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Hepatocellular carcinoma (HCC) is known to be lethal disease. However, its prognosis remains poor, primarily because the precise oncogenic mechanisms underlying HCC progression remain elusive, thus hampering effective treatment. Here, we aimed to identify the potential oncogenes in HCC and elucidate the underlying mechanisms of their action. To identify potential candidate genes, an integrative analysis of eight publicly available genomic datasets was performed, and the functional implications of the identified genes were assessed in vitro and in vivo. Sortilin 1 (SORT1) was identified as a potential candidate oncogene in HCC, and its overexpression in HCC cells was confirmed by analyzing spatial transcriptomic and single-cell data. Silencing SORT1 in Huh-7 and Hep3B cells significantly reduced HCC progression in vitro and in vivo. Functional analyses of oncogenic pathways revealed that SORT1 expression regulated the Notch signaling pathway activation and CD133 expression. Furthermore, analysis of epigenetic regulation of the candidate gene and its clinical implications using The Cancer Genome Atlas Liver Hepatocellular Carcinoma (TCGA LIHC) and our HCC cohort (AJOU_HCC cohort) data demonstrated an inverse correlation between the methylation status of the SORT1 promoter region, specifically at the cg16988986 site, and SORT1 mRNA expression, indicating the epigenetic regulation of SORT1 in HCC. In addition, the distinct methylation status of cg16988986 was significantly associated with patient survival. In conclusion, SORT1 plays a pivotal role in HCC by activating the Notch signaling pathway and increasing CD133 expression. These findings suggest SORT1 as a promising therapeutic target for HCC., (© 2024. The Author(s).)

Background/aims: Hepatocellular carcinoma (HCC) is a leading cause of cancer mortality worldwide. Despite identification of several biomarkers for HCC diagnosis, challenges such as low sensitivity and intratumoral heterogeneity have impeded early detection, highlighting the need for etiology-specific blood biomarkers., Methods: We generated whole-transcriptome sequencing (WTS) and targeted proteome data from buffy coat and plasma samples from HCC patients. By integrating etiological information on viral infection, we investigated the etiology-specific gene expression landscape at the blood level. Validation of differentially expressed genes (DEGs) was performed using publicly available RNA-seq datasets and qRT‒PCR with AUC analyses., Results: Differential expression analyses with multiomics data revealed distinct gene expression profiles between HBV-associated HCC and nonviral HCC, indicating the presence of etiology-specific blood biomarkers. The identified DEGs were validated across multiple independent datasets, underscoring their utility as biomarkers. Additionally, single-cell RNA-seq analysis of HCC confirmed differences in DEG expression across distinct immune cell types., Conclusion: Our buffy coat WTS data and plasma proteome data may serve as reliable sources for identifying etiology-specific blood biomarkers of HCC and might contribute to discovery of therapeutic targets for HCC across different etiologies.

This study evaluated the effect of sodium-glucose cotransporter-2 inhibitors (SGLT2i) on cancer development, particularly in hepatocellular carcinoma (HCC), in individuals with concomitant fatty liver disease (FLD) and type 2 diabetes mellitus (T2DM). Using data from Korea's Health Insurance Review and Assessment Service, we performed Kaplan-Meier and Cox regression analyses in patients with non-alcoholic fatty liver disease (NAFLD) and T2DM (NAFLD-T2DM cohort) and those with chronic viral hepatitis (CVH) alongside FLD and T2DM (FLD-T2DM-CVH cohort). In the propensity score (PS) matched NAFLD-T2DM cohort (N = 107,972), SGLT2i use was not associated with the occurrence of overall cancer, including HCC. However, old age, male sex, liver cirrhosis, and hypothyroidism were identified as independent risk factors for HCC occurrence, whereas statin and fibrate usage were associated with reduced HCC risk in this cohort in multivariate Cox analysis. In the PS-matched FLD-T2DM-CVH cohort (N = 2798), a significant decrease in HCC occurrence was observed among SGLT2i users (P = 0.03). This finding remained consistent in the multivariate Cox regression analysis (Hazard ratio = 2.21, 95% confidence interval = 1.01-4.85, P = 0.048). In conclusion, SGLT2i may be a beneficial option for diabetes management in patients with concomitant T2DM, FLD, and CVH while affirming the overall safety of SGLT2i in other types of cancer., (© 2024. The Author(s).)

Background Noninvasive diagnostic guidelines for hepatocellular carcinoma (HCC) vary across different global geographic areas, especially regarding criteria about gadoxetic acid-enhanced MRI. Purpose To compare the diagnostic performance of four different international HCC diagnosis guidelines and readers' judgment in diagnosing HCC using gadoxetic acid-enhanced MRI in patients at high risk for HCC. Materials and Methods This retrospective study included patients who had not undergone treatment, were at risk for HCC, and who underwent gadoxetic acid-enhanced MRI from January 2015 to June 2018 from 11 tertiary hospitals in South Korea. Four radiologists independently reviewed focal liver lesions (FLLs) according to four guidelines: American Association for the Study of Liver Diseases (AASLD)/Liver Imaging Reporting and Data System (LI-RADS), Korean Liver Cancer Association-National Cancer Center (KLCA-NCC), European Association for the Study of the Liver (EASL), and Asian Pacific Association for the Study of the Liver (APASL). Reader judgment (HCC or not HCC) was also recorded. Malignant FLLs were confirmed at pathology, and histologic and clinical follow-up data were used for benign FLLs. The guidelines' diagnostic performance was compared using generalized estimating equations. Additionally, the diagnostic odds ratio was assessed. Results A total of 2445 FLLs (median size, 27.4 mm) were analyzed in 2237 patients (mean age, 59 years ± 11 [SD]; 1666 male patients); 69.3% (1694 of 2445) were HCCs. KLCA-NCC showed the highest accuracy (80.0%; 95% CI: 78.7, 81.2; P = .001), with high sensitivity in Eastern guidelines (APASL, 89.1% [95% CI: 87.8, 90.3]; KLCA-NCC, 78.2% [95% CI: 76.6, 79.7]) and high specificity in Western guidelines (AASLD/LI-RADS, 89.6% [95% CI: 87.8, 91.2]; EASL, 88.1% [95% CI: 86.2, 89.9]) ( P = .001). The diagnostic odds ratios were 20.7 (95% CI: 17.0, 25.3) for AASLD/LI-RADS, 18.9 (95% CI: 15.8, 22.6) for KLCA-NCC, 16.8 (95% CI: 13.8, 20.4) for EASL, and 8.9 (95% CI: 7.4, 10.7) for APASL. The readers' judgment demonstrated higher accuracy than that of the guidelines (accuracy, 86.0%; 95% CI: 84.9, 86.9; P = .001). Conclusion Among four different international HCC diagnosis guidelines, Eastern guidelines demonstrated higher sensitivity, whereas Western guidelines displayed higher specificity. KLCA-NCC achieved the highest accuracy, and AASLD/LI-RADS exhibited the highest diagnostic odds ratio. © RSNA, 2024 Supplemental material is available for this article.

Background: Patients with Barcelona clinic liver cancer (BCLC) stage B hepatocellular carcinoma (HCC) are considerably heterogeneous in terms of tumor burden, liver function, and performance status. To improve the poor survival outcomes of these patients, treatment approaches other than transarterial chemoembolization (TACE), which is recommended by HCC guidelines, have been adopted in real-world clinical practice. We hypothesize that this non-adherence to treatment guidelines, particularly with respect to the use of liver resection, improves survival in patients with stage B HCC., Aim: To assess guideline adherence in South Korean patients with stage B HCC and study its impact on survival., Methods: A retrospective analysis was conducted using data from 2008 to 2016 obtained from the Korea Central Cancer Registry. Patients with stage B HCC were categorized into three treatment groups, guideline-adherent, upward, and downward, based on HCC guidelines recommended by the Asian Pacific Association for the Study of the Liver (APASL), the European Association for the Study of the Liver (EASL), and the American Association for the Study of Liver Diseases (AASLD). The primary outcome was HCC-related deaths; tumor recurrence served as the secondary outcome. Survival among the groups was compared using the Kaplan-Meier method and the log-rank test. Predictors of survival outcomes were identified using multivariable Cox regression analysis., Results: In South Korea, over the study period from 2008 to 2016, a notable trend was observed in adherence to HCC guidelines. Adherence to the EASL guidelines started relatively high, ranging from 77% to 80% between 2008 and 2012, but it gradually declined to 58.8% to 71.6% from 2013 to 2016. Adherence to the AASLD guidelines began at 71.7% to 75.9% from 2008 to 2010, and then it fluctuated between 49.2% and 73.8% from 2011 to 2016. In contrast, adherence to the APASL guidelines remained consistently high, staying within the range of 90.14% to 94.5% throughout the entire study period. Upward treatment, for example with liver resection, liver transplantation, or radiofrequency ablation, significantly improved the survival of patients with BCLC stage B HCC compared to that of patients treated in adherence to the guidelines (for patients analyzed according to the 2000 EASL guidelines, the 5-year survival rates were 63.4% vs 27.2%, P < 0.001), although results varied depending on the guidelines. Progression-free survival rates were also significantly improved upon the use of upward treatments in certain groups. Patients receiving upward treatments were typically < 70 years old, had platelet counts > 10 5 /μL, and serum albumin levels ≥ 3.5 g/dL., Conclusion: Adherence to guidelines significantly influences survival in South Korean stage B HCC patients. Curative treatments outperform TACE, but liver resection should be selected with caution due to disease heterogeneity., Competing Interests: Conflict-of-interest statement: The authors declare no conflicts of interest., (©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.)

Background: Hepatocellular carcinoma (HCC) accounts for a majority of primary liver cancer cases and related deaths. The purpose of this study was to assess the diagnostic value of splicing factor 3b subunit 4 (SF3B4) as a novel non-invasive biomarker for HCC and determine the association between SF3B4 expression and immune cell infiltration., Methods: An enzyme-linked immunosorbent assay (ELISA) was used to detect SF3B4 levels in plasma samples obtained from healthy controls (HCs) and patients with chronic hepatitis, liver cirrhosis, and HCC. The expression levels of autoantibodies that detect SF3B4 in the plasma samples of each group of patients were measured. Small extracellular vesicles (EVs) were isolated from patient sera, and the expression levels of EV-SF3B4 were measured using quantitative reverse transcription PCR., Results: ELISA results confirmed that the expression levels of SF3B4 proteins and autoantibodies in the plasma of patients with HCC were higher than those in HCs. However, their diagnostic performance was not better than that of alpha-fetoprotein (AFP). The mRNA expression of SF3B4 in serum EV increased but not in the buffy coat or serum of patients with HCC. Serum EV-SF3B4 displayed better diagnostic power than AFP for all stages of HCC (AUC = 0.968 vs. 0.816), including early-stage HCC (AUC = 0.960 vs. 0.842), and this was consistent in the external cohort. Single-cell RNA sequencing indicated that SF3B4 expression was correlated with myeloid-derived suppressor cells. The Tumor Immune Estimation Resource database reconfirmed the correlation between SF3B4 expression and immune cell infiltration in HCC., Conclusions: SF3B4 may be associated with tumor immune infiltration in HCC, and EV-SF3B4 shows potential as a novel non-invasive diagnostic biomarker of HCC., (© 2023. The Author(s).)

Hepatocellular carcinoma (HCC) is a leading cause of cancer-related deaths globally. Early detection and treatment response monitoring of HCC are vital for improved outcomes. Traditional diagnostic methods rely on imaging, serum tumor markers, and invasive liver tissue biopsy. The advent of liquid biopsy has revolutionized cancer diagnostics and management. Liquid biopsy involves the detection and analysis of tumor-derived components, such as circulating tumor DNA, circulating tumor cells, and extracellular vesicles, in various body fluids. These components reflect tumor characteristics, genetic alterations, and functional changes, providing valuable information for HCC diagnosis and treatment response monitoring. Liquid biopsy offers several advantages, including its non-invasive nature, potential for repetitive sampling, and real-time monitoring of disease progression and treatment response. However, challenges remain, including the sensitivity of detection methods, and standardization. In this review, we discuss the methods, current status, and prospects of liquid biopsy for HCC, highlighting its potential as a valuable tool in HCC management.

Background and Aims: This study aimed to compare the long-term cumulative recurrence rates of hepatocellular carcinoma (HCC) and prognosis after curative resection for HCC in noncirrhotic patients with nonalcoholic fatty liver disease (NAFLD) versus hepatitis B virus (HBV) infection., Methods: We retrospectively analyzed the data of 791 patients without recurrence within 1 year after curative resection for HCC from January 2005 to December 2015. Of these, 63 and 728 were NAFLD and HBV patients without cirrhosis, respectively., Results: Recurrence of HCC was observed in 6 (9.5%) and 210 (28.8%) patients in the NAFLD and HBV groups, respectively, during median follow-ups of 69.9 and 85.2 months. Cumulative recurrence rates in the NAFLD group at 2, 4, 6, 8 and 10 years (3.6, 9.4, 12.4, 12.4 and 12.4%, respectively) were significantly lower than in the HBV group (1.7, 16.9, 27.2, 37.1 and 44.4%, respectively) ( P  = 0.008). Cumulative overall survival (OS) rates in the NAFLD group at 2, 4, 6, 8 and 10 years (98.2, 96.0, 84.0, 84.0 and 84.0 %, respectively) were significantly lower than in the HBV group (99.3, 98.4, 97.3, 95.7 and 93.6%, respectively) ( P  = 0.003). HBV infection, with or without fatty liver compared to NAFLD, were significant predictors for the recurrence of HCC ( P  < 0.05 for all) and OS ( P  < 0.05 for all), respectively., Conclusions: Noncirrhotic NAFLD patients showed lower recurrence rates of HCC but poorer survival outcomes than noncirrhotic HBV patients with or without fatty liver. The recurrence risk of HCC remains even in noncirrhotic NAFLD patients., (Copyright © 2022 Wolters Kluwer Health, Inc. All rights reserved.)

Background: Cancer-associated fibroblasts (CAFs) play an important role in the induction of chemo-resistance. This study aimed to clarify the mechanism underlying CAF-mediated resistance to two tyrosine kinase inhibitors (TKIs), sorafenib and lenvatinib, and to identify a novel therapeutic target for overcoming TKI resistance in hepatocellular carcinoma (HCC)., Methods: We performed a systematic integrative analysis of publicly available gene expression datasets and whole-transcriptome sequencing data from 9 pairs of CAFs and para-cancer fibroblasts isolated from human HCC and para-tumor tissues, respectively, to identify key molecules that might induce resistance to TKIs. We then performed in vitro and in vivo experiments to validate selected targets and related mechanisms. The associations of plasma secreted phosphoprotein 1 (SPP1) expression levels before sorafenib/lenvatinib treatment with progression-free survival (PFS) and overall survival (OS) of 54 patients with advanced HCC were evaluated using Kaplan-Meier and Cox regression analysis., Results: Bioinformatic analysis identified CAF-derived SPP1 as a candidate molecule driving TKI resistance. SPP1 inhibitors reversed CAF-induced TKI resistance in vitro and in vivo. CAF-derived SPP1 activated rapidly accelerated fibrosarcoma (RAF)/mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) through the integrin-protein kinase C-alpha (PKCα) signaling pathway and promoted epithelial-to-mesenchymal transition (EMT). A high plasma SPP1 level before TKI treatment was identified as an independent predictor of poor PFS (P = 0.026) and OS (P = 0.047) in patients with advanced HCC after TKI treatment., Conclusions: CAF-derived SPP1 enhances TKI resistance in HCC via bypass activation of oncogenic signals and EMT promotion. Its inhibition represents a promising therapeutic strategy against TKI resistance in HCC. Moreover, plasma SPP1 level before TKI treatment represents a potential biomarker for treatment response prediction., (© 2023 The Authors. Cancer Communications published by John Wiley & Sons Australia, Ltd. on behalf of Sun Yat-sen University Cancer Center.)

Cancer-associated fibroblasts (CAFs) contribute to tumor progression, and microRNAs (miRs) play an important role in regulating the tumor-promoting properties of CAFs. The objectives of this study were to clarify the specific miR expression profile in CAFs of hepatocellular carcinoma (HCC) and identify its target gene signatures. Small-RNA-sequencing data were generated from nine pairs of CAFs and para-cancer fibroblasts isolated from human HCC and para-tumor tissues, respectively. Bioinformatic analyses were performed to identify the HCC-CAF-specific miR expression profile and the target gene signatures of the deregulated miRs in CAFs. Clinical and immunological implications of the target gene signatures were evaluated in The Cancer Genome Atlas Liver Hepatocellular Carcinoma (TCGA&#95;LIHC) database using Cox regression and TIMER analysis. The expressions of hsa-miR-101-3p and hsa-miR-490-3p were significantly downregulated in HCC-CAFs. Their expression in HCC tissue gradually decreased as HCC stage progressed in the clinical staging analysis. Bioinformatic network analysis using miRWalks, miRDB, and miRTarBase databases pointed to TGFBR1 as a common target gene of hsa-miR-101-3p and hsa-miR-490-3p . TGFBR1 expression was negatively correlated with miR-101-3p and miR-490-3p expression in HCC tissues and was also decreased by ectopic miR-101-3p and miR-490-3p expression. HCC patients with TGFBR1 overexpression and downregulated hsa-miR-101-3p and hsa-miR-490-3p demonstrated a significantly poorer prognosis in TCGA&#95;LIHC. TGFBR1 expression was positively correlated with the infiltration of myeloid-derived suppressor cells, regulatory T cells, and M2 macrophages in a TIMER analysis. In conclusion, hsa-miR-101-3p and hsa-miR-490-3p were substantially downregulated miRs in CAFs of HCC, and their common target gene was TGFBR1 . The downregulation of hsa-miR-101-3p and hsa-miR-490-3p , as well as high TGFBR1 expression, was associated with poor clinical outcome in HCC patients. In addition, TGFBR1 expression was correlated with the infiltration of immunosuppressive immune cells.

Purpose: To investigate added value of late portal venous phase (LPVP) for identification of enhancing capsule (EC) on gadoxetate disodium-enhanced MRI (GD-MRI) for diagnosing hepatocellular carcinoma (HCC) in patients with chronic liver disease (CLD)., Methods: This retrospective study comprised 116 high-risk patients with 128 pathologically proven HCCs who underwent GD-MRI including arterial phase, conventional portal venous phase (CPVP, 60 s), LPVP (mean, 104.4 ± 6.7 s; range, 90-119 s), and transitional phase (TP, 3 min). Two independent radiologists assessed the presence of major HCC features, including EC on CPVP and/or TP (CPVP/TP) and EC on LPVP. The frequency of EC was compared on GD-MRI between with and without inclusion of LPVP. The radiologists assigned Liver Imaging Reporting and Data System (LI-RADS) v2018 categories before and after identifying EC on LPVP., Results: Of the total 128 HCCs, 74 and 73 revealed EC on CPVP/TP for reviewer 1 and 2, respectively. After inclusion of LPVP, each reviewer identified seven more EC [Reviewer 1, 57.8% (74/128) vs. 63.3% (81/128); Reviewer 2, 57.0% (73/128) vs. 62.5% (80/128); P = 0.016, respectively]. Sensitivities of LR-5 assignment for diagnosing HCCs were not significantly different in GD-MRI with or without LPVP for EC identification [Reviewer 1, 71.9% (92/128) vs. 72.7% (93/128); Reviewer 2, 75.0% (96/128) vs. 75.8% (97/128); P = 1.000, respectively]., Conclusion: Including the LPVP in GD-MRI may improve identification of EC of HCC in patients with CLD. However, LI-RADS v2018 using GD-MRI showed comparable sensitivity for diagnosing HCC regardless of applying LPVP for EC., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Objectives: We aimed to investigate the optimal radiologic method to determine Milan criteria (MC) for the prediction of recurrence in patients who underwent locoregional treatment (LRT) for hepatocellular carcinoma (HCC) and subsequent liver transplantation (LT)., Methods: This retrospective study included 121 HCC patients who underwent LRT and had both liver dynamic CT and MRI. They were classified with MC using four cross combinations of two imaging modalities (CT and MRI) and two diagnostic criteria (modified Response Evaluation Criteria in Solid Tumors [mRECIST] and Liver Imaging Reporting and Data System treatment response algorithm [LI-RADS TRA]). Competing risk regression was performed to analyze the time to recurrence after LT. The predictive abilities of the four methods for recurrence were evaluated using the time-dependent area under the curve (AUC)., Results: Competing risk regression analyses found that beyond MC determined by MRI with mRECIST was independently associated with recurrence (hazard ratio, 6.926; p = 0.001). With mRECIST, MRI showed significantly higher AUCs than CT at 3 years and 5 years after LT (0.597 vs. 0.756, p = 0.012 at 3 years; and 0.588 vs. 0.733, p = 0.024 at 5 years). Using the pathologic reference standard, MRI with LI-RADS TRA showed higher sensitivity (61.5%) than CT with LI-RADS TRA (30.8%, p < 0.001) or MRI with mRECIST (38.5%, p < 0.001)., Conclusions: MRI with mRECIST was the optimal radiologic method to determine MC for the prediction of post-LT recurrence in HCC patients with prior LRT., Key Points: • MRI with modified RECIST (mRECIST) is the optimal preoperative method to determine Milan criteria for the prediction of post-transplant HCC recurrence in patients with prior locoregional treatment. • With mRECIST, MRI was better than CT for the prediction of post-transplant recurrence., (© 2022. The Author(s), under exclusive licence to European Society of Radiology.)

Background/aims: A comprehensive analysis of trends in the incidence of hepatocellular carcinoma (HCC) is important for planning public health initiatives. We aimed to analyze the trends in HCC incidence in South Korea over 10 years and to predict the incidence for the year 2028., Methods: Data from patients with newly diagnosed HCC between 2008 and 2018 were obtained from Korean National Health Insurance Service database. Age-standardized incidence rates (ASRs) were calculated to compare HCC incidence. A poisson regression model was used to predict the future incidence of HCC., Results: The average crude incidence rate (CR) was 22.4 per 100,000 person-years, and the average ASR was 17.6 per 100,000 person-years between 2008 and 2018. The CR (from 23.9 to 21.2 per 100,000 person-years) and ASR (from 21.9 to 14.3 per 100,000 person-years) of HCC incidence decreased during the past ten years in all age groups, except in the elderly. The ASR of patients aged ≥80 years increased significantly (from 70.0 to 160.2/100,000 person-years; average annual percent change, +9.00%; P<0.001). The estimated CR (17.9 per 100,000 person-years) and ASR (9.7 per 100,000 person-years) of HCC incidence in 2028 was declined, but the number of HCC patients aged ≥80 years in 2028 will be quadruple greater than the number of HCC patients in 2008 (from 521 to 2,055), comprising 21.3% of all HCC patients in 2028., Conclusion: The ASRs of HCC in Korea have gradually declined over the past 10 years, but the number, CR, and ASR are increasing in patients aged ≥80 years.

Non-alcoholic fatty liver disease (NAFLD), attributed to excessive fat accumulation in the liver, is reportedly prevalent worldwide. NAFLD is one of the leading causes of chronic liver disease, including non-alcoholic steatohepatitis (NASH), cirrhosis, and hepatic cellular carcinoma (HCC). The peripheral roles of serotonin (5-hydroxytryptamine, 5HT) were found to regulate hepatic lipid metabolism. Among serotonin receptor subtypes, 5HT 2A receptor is known to regulate hepatic lipid metabolism. Hepatic lipid accumulation and hepatic triglyceride (TG) were reduced in liver-specific 5HT 2A receptor knockout (5HT 2A receptor LKO) mice upon high-fat diet (HFD) feeding. In the present study, we explored a series of new peripherally acting 5HT 2A receptor antagonists. Compound 14a displayed good in vitro activity, with an IC 50 value of 0.17 nM. Compound 14a exhibited good microsomal stability, no significant CYP and hERG inhibition, and 5HT receptor subtype selectivity. The brain-to-plasma ratio of 14a was below the lower limit of quantification, indicating limited blood-brain barrier (BBB) penetration. HFD-fed 14a treated mice showed decreased liver steatosis and lobular inflammation. These results demonstrate the potential of newly synthesized peripheral 5HT 2A receptor antagonists for treating NAFLD., (Copyright © 2022. Published by Elsevier Masson SAS.)

BACKGROUND. In LI-RADS version 2018, observations showing at least one of five targetoid appearances in different sequences or postcontrast phases are categorized LR-M, indicating likely non-hepatocellular carcinoma (HCC) malignancy. OBJECTIVE. The purpose of this study was to evaluate interobserver agreement for LI-RADS targetoid appearances among a large number of radiologists of varying experience and the diagnostic performance of targetoid appearances for differentiating HCC from non-HCC malignancy. METHODS. This retrospective study included 100 patients (76 men, 24 women; mean age, 58 ± 9 [SD] years) at high risk of HCC who underwent gadoxetic acid-enhanced MRI within 30 days before hepatic tumor resection (25 randomly included patients with non-HCC malignancy [13, intrahepatic cholangiocarcinoma; 12, combined HCC-cholangiocarcinoma]; 75 matched patients with HCC). Eight radiologists (four more experienced [8-15 years]; four less experienced [1-5 years]) from seven institutions independently assessed observations for the five targetoid appearances and LI-RADS categorization. Interobserver agreement and diagnostic performance for non-HCC malignancy were evaluated. RESULTS. Interobserver agreement was poor for peripheral washout (κ = 0.20); moderate for targetoid transitional phase or hepatobiliary phase appearance (κ = 0.33), delayed central enhancement (κ = 0.37), and targetoid restriction (κ = 0.43); and substantial for rim arterial phase hyperenhancement (κ = 0.61). Agreement was fair for at least one targetoid appearance (κ = 0.36) and moderate for at least two, three, or four targetoid appearances (κ = 0.43-0.51). Agreement for individual targetoid appearances was not significantly different between more experienced and less experienced readers other than for targetoid restriction (κ = 0.63 vs 0.43; p = .001). Agreement for at least one targetoid appearance was fair among more experienced (κ = 0.29) and less experienced (κ = 0.37) reviewers. Agreement for at least two, three, or four targetoid appearances was moderate to substantial among more experienced reviewers (κ = 0.45-0.63) and moderate among less experienced reviewers (κ = 0.42-0.56). Existing LR-M criteria of at least one targetoid appearance had median accuracy for non-HCC malignancy of 62%, sensitivity of 84%, and specificity of 54%. For all reviewers, accuracy was highest when at least three (median accuracy, 79%; sensitivity, 68%; specificity, 82%) or four (median accuracy, 80%; sensitivity, 54%; specificity, 88%) targetoid appearances were required. CONCLUSION. Targetoid appearances and LR-M categorization exhibited considerable interobserver variation among both more and less experienced reviewers. CLINICAL IMPACT. Requiring multiple targetoid appearances for LR-M categorization improved interobserver agreement and diagnostic accuracy for non-HCC malignancy.

For hepatocellular carcinoma, the IMbrave150 trial demonstrated that combination atezolizumab and bevacizumab had significantly better overall survival compared with sorafenib in patients with unresectable disease. However, as more immune checkpoint inhibitors are approved as first-line agents in gastrointestinal cancers, there have been few reports on whether sequential PD-1/PD-L1 blockade is beneficial in the treatment of these diseases. We present a patient with hepatocellular carcinoma who had disease progression on atezolizumab, a PD-L1 inhibitor, but subsequently had a remarkable response to pembrolizumab, a PD-1 inhibitor., Competing Interests: Competing interests: None declared., (© BMJ Publishing Group Limited 2022. No commercial re-use. See rights and permissions. Published by BMJ.)

Background: Hepatocellular carcinoma (HCC) is one of the most common and lethal cancers worldwide. Wiskott-Aldrich syndrome protein family member 2 (WASF2) is an integral member of the actin cytoskeleton pathway, which plays a crucial role in cell motility. In this study, we aimed to explore the role of WASF2 in HCC carcinogenesis and its regulatory mechanism., Methods: WASF2 expression in HCC was analyzed using six public RNA-seq datasets and 66 paired tissues from patients with HCC. The role of WASF2 in normal hepatocyte cell phenotypes was evaluated using a WASF2 overexpression vector in vitro; it was evaluated in HCC cell phenotypes using small interfering RNA (siRNA) in vitro and in vivo. Epigenetic regulatory mechanism of WASF2 was assessed in the Cancer Genome Atlas liver hepatocellular carcinoma project (TCGA&#95;LIHC) dataset and also validated in 38 paired HCC tissues. Site mutagenesis, bisulfite sequencing polymerase chain reaction (BSP), methylation-specific polymerase chain reaction (MSP), and quantitative MSP (qMSP) were used for evaluating WASF2 methylation status., Results: WASF2 is overexpressed in HCC and is clinically correlated with its prognosis. WASF2 overexpression promoted normal hepatocyte proliferation. WASF2 inactivation decreased the viability, growth, proliferation, migration, and invasion of Huh-7 and SNU475 HCC cells by inducing G2/M phase arrest. This induced cell death and inhibited epithelial-mesenchymal transition, hindering actin polymerization. In addition, WASF2 knockdown using siWASF2 in a xenograft mouse model and a lung metastasis model exerted tumor suppressive effect. There was a negative correlation between WASF2 methylation status and mRNA expression. The methylation pattern of CpG site 2 (- 726 bp), located in the WASF2 promoter, plays an important role in the regulation of WASF2 expression. Furthermore, the cg242579 CpG island in the WASF2 5' promoter region was hypomethylated in HCC compared to that in the matched non-tumor samples. Patients with high WASF2 methylation and low WASF2 expression displayed the highest overall survival., Conclusions: WASF2 is overexpressed and hypomethylated in HCC and correlates with patient prognosis. WASF2 inactivation exerts anti-tumorigenic effects on HCC cells in vitro and in vivo, suggesting that WASF2 could be a potential therapeutic target for HCC., (© 2022. The Author(s).)

Background/aims: Posthepatectomy liver failure (PHLF) is a major complication that increases mortality in patients with hepatocellular carcinoma after surgical resection. The aim of this retrospective study was to evaluate the utility of magnetic resonance elastography-assessed liver stiffness (MRE-LS) for the prediction of PHLF and to develop an MRE-LS-based risk prediction model., Methods: A total of 160 hepatocellular carcinoma patients who underwent surgical resection with available preoperative MRE-LS data were enrolled. Clinical and laboratory parameters were collected from medical records. Logistic regression analyses were conducted to identify the risk factors for PHLF and develop a risk prediction model., Results: PHLF was present in 24 patients (15%). In the multivariate logistic analysis, high MRE-LS (kPa; odds ratio [OR] 1.49, 95% confidence interval [CI] 1.12 to 1.98, p=0.006), low serum albumin (≤3.8 g/dL; OR 15.89, 95% CI 2.41 to 104.82, p=0.004), major hepatic resection (OR 4.16, 95% CI 1.40 to 12.38, p=0.014), higher albumin-bilirubin score (>-0.55; OR 3.72, 95% CI 1.15 to 12.04, p=0.028), and higher serum α-fetoprotein (>100 ng/mL; OR 3.53, 95% CI 1.20 to 10.40, p=0.022) were identified as independent risk factors for PHLF. A risk prediction model for PHLF was established using the multivariate logistic regression equation. The area under the receiver operating characteristic curve (AUC) of the risk prediction model was 0.877 for predicting PHLF and 0.923 for predicting grade B and C PHLF. In leave-one-out cross-validation, the risk model showed good performance, with AUCs of 0.807 for all-grade PHLF and 0. 871 for grade B and C PHLF., Conclusions: Our novel MRE-LS-based risk model had excellent performance in predicting PHLF, especially grade B and C PHLF.

Fibrolamellar carcinoma (FLC) is a primary liver cancer that most commonly arises in adolescents and young adults in a background of normal liver tissue and has a poor prognosis due to lack of effective chemotherapeutic agents. The DNAJB1-PRKACA gene fusion (DP) has been reported in the majority of FLC tumors; however, its oncogenic mechanisms remain unclear. Given the paucity of cellular models, in particular FLC tumor cell lines, we hypothesized that engineering the DP fusion gene in HEK293T cells would provide insight into the cellular effects of the fusion gene. We used CRISPR/Cas9 to engineer HEK293T clones expressing DP fusion gene (HEK-DP) and performed transcriptomic, proteomic, and mitochondrial studies to characterize this cellular model. Proteomic analysis of DP interacting partners identified mitochondrial proteins as well as proteins in other subcellular compartments. HEK-DP cells demonstrated significantly elevated mitochondrial fission, which suggests a role for DP in altering mitochondrial dynamics. Transcriptomic analysis of HEK-DP cells revealed a significant increase in LINC00473 expression, similar to what has been observed in primary FLC samples. LINC00473 overexpression was reversible with siRNA targeting of PRKACA as well as pharmacologic targeting of PKA and Hsp40 in HEK-DP cells. Therefore, our model suggests that LINC00473 is a candidate marker for DP activity., Competing Interests: The authors have declared that no competing interests exist.

Background: According to the Liver Imaging Reporting and Data System (LI-RADS), the LI-RADS category M (LR-M), which are probably or definitely malignant but are not specific for hepatocellular carcinomas (HCCs), does not exclude HCCs. A gap in knowledge remains, including their characteristics and recurrence of HCCs categorized as LR-M., Purpose: To compare the characteristics of HCCs categorized as LR-M with HCCs categorized as LR-4 or LR-5 (LR-4/5) using the LI-RADS version 2018 and evaluate the relationship of these categories with the risk of early recurrence after curative resections of single HCCs., Study Type: Retrospective., Subjects: Two hundred and eighty-one patients (mean age, 57 years; 191 men and 90 women) who underwent curative resections for single HCCs and preoperative contrast-enhanced MRI between 2015 and 2017., Field Strength/sequence: 3T Dual gradient-echo T 1 WI with in- and opposed-phase, turbo spin-echo T 2 WI, diffusion-weighted echo-planar images, and three-dimensional gradient-echo T 1 WI before and after administration of contrast agent., Assessment: MRI features according to the LI-RADS version 2018 were evaluated and LI-RADS category were assigned for each observation. Clinical, imaging, and histopathological features were compared based on LI-RADS categorization. Early recurrence rates (<2 years) and associated factors were also evaluated., Statistical Tests: Fisher's exact test, two-sample t test after satisfying assumption of normality through Shapiro-Wilk test, Fleiss κ coefficient, Cox proportional hazards regression analysis, Kaplan-Meier method, and log-rank test., Results: Forty-one HCCs (14.6%) were categorized as LR-M and 240 HCCs (85.4%) were categorized as LR-4/5. LR-M HCCs showed poorer differentiation than LR-4/5 HCCs. In the multivariate analysis, the LR-M category was an independent predictor for early recurrence (hazard ratio, 1.904; 95% confidence interval, 1.024-3.542; P < 0.05). Early recurrence rates were significantly higher in patients with LR-M HCCs than in patients with LR-4/5 HCCs (32.0% vs. 18.4%, respectively, P < 0 05)., Data Conclusion: Compared to LR-4/5 HCCs, LR-M HCCs were associated with poorer tumor differentiation and higher early recurrence rates after curative resections of single HCCs., Level of Evidence: 3 Technical Efficacy Stage: 2., (© 2021 International Society for Magnetic Resonance in Medicine.)

This study investigated the diagnostic potential of serum small extracellular vesicle-derived long noncoding RNAs (EV-lncRNAs) for hepatocellular carcinoma (HCC). Driver oncogenic lncRNA candidates were selected by a comparative analysis of lncRNA expression profiles from two whole transcriptome human HCC datasets (Catholic&#95;LIHC and TCGA&#95;LIHC). Expression of selected lncRNAs in serum and small EVs was evaluated using quantitative reverse transcription PCR. Diagnostic power of serum EV-lncRNAs for HCC was determined in the test (n = 44) and validation (n = 139) cohorts. Of the six promising driver onco-lncRNAs, DLEU2, HOTTIP, MALAT1, and SNHG1 exhibited favorable performance in the test cohort. In the validation cohort, serum EV-MALAT1 displayed excellent discriminant ability, while EV-DLEU2, EV-HOTTIP, and EV-SNHG1 showed good discriminant ability between HCC and non-HCC. Furthermore, a panel combining EV-MALAT1 and EV-SNHG1 achieved the best area under the curve (AUC; 0.899, 95% CI = 0.816-0.982) for very early HCC, whereas a panel with EV-DLEU2 and alpha-fetoprotein exhibited the best positivity (96%) in very early HCC. Serum small EV-MALAT1, EV-DLEU2, EV-HOTTIP, and EV-SNHG1 may represent promising diagnostic markers for very early-stage HCC., (© 2021 The Authors. Molecular Oncology published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.)

Objective: To assess the diagnostic performance of the Liver Imaging Reporting and Data System (LI-RADS) version 2018 treatment response algorithm (TRA) for the evaluation of hepatocellular carcinoma (HCC) treated with transarterial radioembolization., Materials and Methods: This retrospective study included patients who underwent transarterial radioembolization for HCC followed by hepatic surgery between January 2011 and December 2019. The resected lesions were determined to have either complete (100%) or incomplete (< 100%) necrosis based on histopathology. Three radiologists independently reviewed the CT or MR images of pre- and post-treatment lesions and assigned categories based on the LI-RADS version 2018 and the TRA, respectively. Diagnostic performances of LI-RADS treatment response (LR-TR) viable and nonviable categories were assessed for each reader, using histopathology from hepatic surgeries as a reference standard. Inter-reader agreements were evaluated using Fleiss κ., Results: A total of 27 patients (mean age ± standard deviation, 55.9 ± 9.1 years; 24 male) with 34 lesions (15 with complete necrosis and 19 with incomplete necrosis on histopathology) were included. To predict complete necrosis, the LR-TR nonviable category had a sensitivity of 73.3-80.0% and a specificity of 78.9-89.5%. For predicting incomplete necrosis, the LR-TR viable category had a sensitivity of 73.7-79.0% and a specificity of 93.3-100%. Five (14.7%) of 34 treated lesions were categorized as LR-TR equivocal by consensus, with two of the five lesions demonstrating incomplete necrosis. Inter-reader agreement for the LR-TR category was 0.81 (95% confidence interval: 0.66-0.96)., Conclusion: The LI-RADS version 2018 TRA can be used to predict the histopathologic viability of HCCs treated with transarterial radioembolization., Competing Interests: The authors have no potential conflicts of interest to disclose., (Copyright © 2021 The Korean Society of Radiology.)

Background and Aims: The influence of direct-acting antivirals (DAAs) on chronic hepatitis C (CHC)-related hepatocellular carcinoma (HCC) remains controversial. We investigated the effect of eradicating CHC using DAAs on treatment outcomes in patients with CHC-related HCC treated with transarterial chemoembolization (TACE)., Methods: This nationwide, multi-center, retrospective study recruited patients with CHC-related HCC treated with TACE as the first-line anti-cancer treatment, and who achieved a sustained virological response (SVR) using DAAs (DAA group) between 2006 and 2017. Patients achieving an SVR following interferon-based treatment (IFN group) and those without treatment (control group) were also recruited for comparison., Results: A total of 425 patients were eligible for the study. Of these, 356 (83.8%), 26 (6.1%), and 43 (10.1%) were allocated to the control, IFN, and DAA groups, respectively. A multivariate analysis showed that liver cirrhosis, segmental portal vein thrombosis, and larger maximal tumor size independently predicted an increased risk of progression (all p < 0.05), whereas, the DAA group (vs. IFN and control groups) independently predicted a reduced risk of progression (hazard ratio (HR) = 0.630, 95% confidence interval 0.411-0.966, p = 0.034). The cumulative incidence rate of HCC progression in the DAA group was significantly lower than that in the IFN and control groups (p = 0.033, log-rank test). In addition, the DAA group (vs. IFN and control groups) was independently associated with a reduced risk of mortality (p = 0.042)., Conclusions: DAA treatment provided significantly prolonged progression-free survival in patients with CHC-related HCC treated with TACE compared to that in patients administered IFN or no treatment.

Background/aims: This study was performed to evaluate the efficacy of direct-acting antivirals (DAAs) in Korean patients with hepatitis C virus (HCV)-related hepatocellular carcinoma (HCC) and to investigate the risk factors associated with HCC recurrence., Methods: A total of 100 patients with HCV-related HCC, who were treated with DAAs between May 2015 and December 2016, were recruited from seven university hospitals in Korea. Claim data of 526 patients with HCC obtained from the Health Insurance Review and Assessment Service in South Korea were used for external validation of the results., Results: Among the 100 patients, 88% achieved a sustained virological response (SVR) 12 weeks after the end of DAA therapy (SVR12), and 37% experienced HCC recurrence after DAA therapy. Short last HCC treatment durability (<12 months) before DAA commencement was independently associated with HCC recurrence (hazard ratio [HR], 2.89; p=0.011). In the nationwide validation cohort, 20.3% of the patients experienced HCC recurrence. The last HCC treatment with a noncurative method, a short last HCC treatment durability (<12 months), and a longer total duration of HCC treatment (≥18 months) were independently related with HCC recurrence (HR 3.73, p<0.001; HR 3.34, p<0.001; and HR 1.74, p=0.006; respectively)., Conclusions: DAA therapy showed an acceptable SVR12 rate in patients with HCV-related HCC. Short last HCC treatment durability (<12 months) was associated with HCC recurrence after DAA therapy. This finding suggests that the last HCC treatment durability is an important predictor of HCC recurrence after DAA therapy.

Objectives: The primary aim of this study was to determine the clinical and histopathological prognostic factors for patients who underwent surgical resection of multiple hepatocellular carcinomas (HCCs) of multicentric occurrence. The secondary aim of this study was to evaluate whether specific imaging-related factors, including arterial phase hyperenhancement (APHE) and the LI-RADS category of each lesion on gadoxetic acid-enhanced MRI, would provide additional prognostic information about multicentric HCCs., Methods: In this retrospective study, 54 patients with 120 multicentric HCCs were diagnosed by surgical resection at a single tertiary hospital between 2009 and 2014. Two independent readers evaluated patients' preoperative gadoxetic acid-enhanced MR images and recorded APHE and LI-RADS category for each HCC, with discrepancies resolved through consensus sessions if necessary. Potential clinicopathologic and imaging parameters for predicting disease-free survival (DFS) and overall survival (OS) were analyzed using Cox regression analysis., Results: Presence of microvascular invasion (MVI) (p = 0.003) and of three or more HCCs (p = 0.013) were both independent predictors of a shorter DFS. Patients with concurrent MVI and three or more HCCs had the shortest DFS. MVI was the only statistically significant parameter (p = 0.023) predicting OS. The number of HCCs with APHE or LR-5/M category was not associated with survival., Conclusions: Presence of MVI and of three or more HCCs were associated with poorer outcomes after surgical resection of multicentric HCCs. Imaging parameters on gadoxetic acid-enhanced MRI such as APHE or LI-RADS category were not associated with postsurgical outcomes., Key Points: • Patients with three or more hepatocellular carcinomas showed worse disease-free survival than those with two hepatocellular carcinomas after surgical resection. • Microvascular invasion was the only significant factor to affect both the disease-free and overall survivals of patients after surgical resection of multicentric hepatocellular carcinomas. • Preoperative MRI findings related to multicentric hepatocellular carcinomas such as arterial phase hyperenhancement and LI-RADS category of lesions did not provide significant prognostic information.

Objective: We investigated whether liver stiffness (LS) quantified using magnetic resonance elastography (MRE) could predict the prognosis of advanced hepatocellular carcinoma (HCC) patients treated with sorafenib., Methods: We selected 50 sorafenib-treated advanced HCC patients who underwent MRE within 3 months before drug administration from a prospectively maintained cohort of chronic liver disease patients, according to the inclusion and exclusion criteria. Univariate and multivariate analyses were performed to evaluate the prognostic role of laboratory data, tumor characteristics, and MRE-assessed LS for overall survival (OS), progression-free survival (PFS), and significant liver injury (grade ≥ 3) after sorafenib administration., Results: High MRE-assessed LS either as continuous (per kPa, hazard ratio (HR) 1.54; 95% confidence interval (CI) 1.23-1.92, p < 0.001) or categorical (> 7.5 kPa, HR 4.06, 95% CI 1.40-11.79, p < 0.01) variable was significantly associated with poor OS along with higher serum alpha-fetoprotein (AFP, ≥ 400 ng/mL) and advanced tumor stage (modified Union for International Cancer Control (mUICC) IVb). Higher MRE-assessed LS was also significantly associated with the development of significant liver injury after sorafenib administration (per kPa, HR 1.62, 95% CI 1.21-2.17, p = 0.001; > 7.5 kPa, HR 10.11, 95% CI 2.41-42.46, p = 0.002). PFS analysis identified higher serum AFP (≥ 400 ng/mL) and advanced tumor stage (mUICC IVb) as significant risk factors for early disease progression, whereas LS was not associated with PFS CONCLUSION: Higher MRE-assessed LS is a potential biomarker for predicting poor OS and significant liver injury in advanced HCC patients treated with sorafenib., Key Points: • Higher pretreatment LS by MRE (> 7.5 kPa), higher AFP (≥ 400 ng/mL), and advanced tumor stage (mUICC IVb) were associated with poor OS in advanced HCC patients treated with sorafenib. • Higher pretreatment LS by MRE was associated with developing significant (grade ≥ 3) liver injury during sorafenib treatment, which required termination of the therapy. • Patients with high pretreatment LS by MRE should be monitored carefully for potential liver injury during sorafenib treatment.

Objectives: This study aimed to assess the detection of hepatocellular carcinoma (HCC) utilizing subtraction AP (arterial phase) imaging only for T1 hyperintense observations compared with the detection of HCC on subtraction AP imaging that included T1 hyper-, iso-, and hypointense lesions on gadoxetate disodium-enhanced MRI., Materials and Methods: This retrospective study included 234 patients (311 observations including 239 HCCs) at high risk for HCC who underwent gadoxetate disodium-enhanced MRI with subtraction AP imaging between 2015 and 2017. Arterial phase hyperenhancement (APHE) was divided into two subtypes: conventional APHE, where subtraction AP imaging is used to detect APHE only for T1 hyperintense observations; and modified APHE, where subtraction AP imaging is applied to T1 hyper-, iso-, and hypointense lesions. Two readers independently reviewed all observations and the per-observation diagnostic performances were compared using McNemar's test., Results: Modified nonrim APHE showed significantly higher sensitivity than conventional nonrim APHE (90.0% vs 82.8%; p < 0.001) for diagnosing HCC, without a significant difference in specificity (66.7% vs 68.1%; p > 0.999). The LR-5 category with modified nonrim APHE provided better sensitivity than the LR-5 with conventional nonrim APHE (70.3% vs 63.2%; p < 0.001), without a significant decrease in specificity (94.4% vs 95.8%; p > 0.999)., Conclusion: Extended application of subtraction AP imaging for T1 hypo- or isointense observations on gadoxetate disodium-enhanced MRI can improve sensitivity in the diagnosis of HCC without a significant difference in specificity., Key Points: • Modified nonrim arterial phase hyperenhancement (APHE), extended application of subtraction arterial phase imaging for T1 hypo- or isointense observation, outperforms conventional nonrim APHE. • The LR-5 category with modified nonrim APHE provided better sensitivity in diagnosing HCC than the LR-5 with conventional APHE, without a significant decrease in specificity.

Objectives: The current LR-5 criteria of Liver Imaging Reporting and Data System (LI-RADS) determined by only major features provide high specificity, but unsatisfactorily low sensitivity for the noninvasive diagnosis of hepatocellular carcinoma (HCC). This study aimed to identify significant ancillary features (AFs) in LI-RADS version 2018 and develop the upgraded LR-5 criteria to improve diagnostic performance on gadoxetic acid-enhanced magnetic resonance imaging (MRI)., Methods: This retrospective study included 280 patients (366 observations including 281 HCCs) at high-risk for HCC who underwent gadoxetic acid-enhanced MRI between 2015 and 2017. Two readers evaluated major features and AFs for each observation and assigned a LI-RADS category. Independently significant AFs were identified through logistic regression analysis. Upgraded LR-5 criteria were developed by combining independently significant AFs with LR-4 assigned by major features alone. Sensitivities and specificities of the diagnostic criteria were compared using McNemar's test., Results: Two of the AFs favoring malignancy in general (mild-moderate T2 hyperintensity and hepatobiliary phase hypointensity) and two of the AFs favoring HCC in particular (nonenhancing "capsule" and mosaic architecture) were independently significant features for diagnosing HCC. By using the upgraded LR-5 criteria (LR-4 by major features alone + each aforementioned AF), sensitivities were significantly increased (69.4-76.9%) compared with the standard LR-5 (66.2%; all, p ≤ 0.004), whereas specificities (95.3-96.5%) were not significantly different (96.5%; all, p > 0.999)., Conclusions: Independently significant AFs may be used to upgrade from LR-4 to LR-5 to improve sensitivity without impairing specificity on gadoxetic acid-enhanced MRI., Key Points: • Independently significant AFs for HCC on gadoxetic acid-enhanced MRI were mild-moderate T2 hyperintensity, hepatobiliary phase hypointensity, nonenhancing "capsule," and mosaic architecture. • When LR-4 criteria by major features alone in combination with significant AFs were upgraded to LR-5, sensitivities were higher than the standard LR-5, without impairing specificity. • Independently significant ancillary features in Liver Imaging Reporting and Data System version 2018 may be used to upgrade from LR-4 to LR-5 to improve sensitivity without impairing specificity on gadoxetic acid-enhanced MRI.

Purpose: Alpha-fetoprotein (AFP) is a prognostic marker for hepatocellular carcinoma (HCC). We investigated the prognostic value of AFP levels in patients who achieved complete response (CR) to transarterial chemoembolization (TACE) for HCC., Materials and Methods: Between 2005 and 2018, 890 patients with HCC who achieved a CR to TACE were recruited. An AFP responder was defined as a patient who showed elevated levels of AFP (>10 ng/mL) during TACE, but showed normalization or a >50% reduction in AFP levels after achieving a CR., Results: Among the recruited patients, 569 (63.9%) with naïve HCC and 321 (36.1%) with recurrent HCC after complete resection were treated. Before TACE, 305 (34.3%) patients had multiple tumors, 219 (24.6%) had a maximal tumor size >3 cm, and 22 (2.5%) had portal vein tumor thrombosis. The median AFP level after achieving a CR was 6.36 ng/mL. After a CR, 473 (53.1%) patients experienced recurrence, and 417 (46.9%) died [median progression-free survival (PFS) and overall survival (OS) of 16.3 and 62.8 months, respectively]. High AFP levels at CR (>20 ng/mL) were independently associated with a shorter PFS [hazard ratio (HR)=1.403] and OS (HR=1.284), together with tumor multiplicity at TACE (HR=1.518 and 1.666, respectively). AFP non-responders at CR (76.2%, n=359 of 471) showed a shorter PFS (median 10.5 months vs. 15.5 months, HR=1.375) and OS (median 41.4 months vs. 61.8 months, HR=1.424) than AFP responders (all p =0.001)., Conclusion: High AFP levels and AFP non-responders were independently associated with poor outcomes after TACE. AFP holds clinical implications for detailed risk stratification upon achieving a CR after TACE., Competing Interests: The authors have no potential conflicts of interest to disclose., (© Copyright: Yonsei University College of Medicine 2021.)

Purpose: To evaluate the diagnostic performance of LR-M criteria for differentiating hepatocellular carcinoma, intrahepatic mass-forming cholangiocarcinoma, and combined hepatocellular-cholangiocarcinoma and to compare the imaging features of each type., Methods: In this retrospective study, 110 patients were surgically diagnosed with cholangiocarcinoma (n = 67) and combined hepatocellular-cholangiocarcinoma (n = 43) at a single tertiary hospital between 2013 and 2018. Among them, those with risk factors were enrolled (16 cholangiocarcinomas and 33 combined hepatocellular-cholangiocarcinomas). Forty-nine other patients with size-matched hepatocellular carcinoma were selected as a control group. Two independent readers evaluated the imaging findings of the preoperative MRIs based on LI-RADS version 2018 and assigned an LI-RADS category. The diagnostic performance of the LR-M criteria for diagnosing cholangiocarcinoma or combined hepatocellular-cholangiocarcinoma was evaluated, and the imaging features were compared. The imaging findings of the tumors in patients without risk factors (51 cholangiocarcinomas and 10 combined hepatocellular-cholangiocarcinomas) were evaluated for subgroup analysis., Results: In the non-hepatocellular carcinoma group, 33 patients were categorized into LR-M and 14 patients into LR-5 (67.3% and 28.6%, respectively), while 5 patients with hepatocellular carcinoma were categorized into LR-M and 38 patients into LR-5 (10.2% and 77.6%, respectively). Sensitivity and specificity of the LR-M criteria were 67.3% and 89.8%, respectively. When more than two LR-M features were present, cholangiocarcinoma or combined hepatocellular-cholangiocarcinoma were suggested with a specificity of 95.9%., Conclusion: The diagnostic performance of the LR-M criteria is acceptable with moderate sensitivity and high specificity for both cholangiocarcinoma and combined hepatocellular-cholangiocarcinoma. Imaging findings of primary hepatic carcinomas should be understood as a spectrum.

Liquid biopsy can provide a strong basis for precision medicine. We aimed to identify novel single-nucleotide variants (SNVs) in circulating tumor DNA (ctDNA) in patients with hepatocellular carcinoma (HCC). Deep sequencing of plasma-derived ctDNA from 59 patients with HCC was performed using a panel of 2924 SNVs in 69 genes. In 55.9% of the patients, at least one somatic mutation was detected. Among 25 SNVs in 12 genes, four frequently observed SNVs, MLH1 (13%), STK11 (13%), PTEN (9%), and CTNNB1 (4%), were validated using droplet digital polymerase chain reaction with ctDNA from 62 patients with HCC. Three candidate SNVs were detected in 35.5% of the patients, with a frequency of 19% for MLH1 chr3:37025749T>A, 11% for STK11 chr19:1223126C>G, and 8% for PTEN chr10:87864461C>G. The MLH1 and STK11 SNVs were also confirmed in HCC tissues. The presence of the MLH1 SNV, in combination with an increased ctDNA level, predicted poor overall survival among 107 patients. MLH1 chr3:37025749T>A SNV detection in ctDNA is feasible, and thus, ctDNA can be used to detect somatic mutations in HCC. Furthermore, the presence or absence of the MLH1 SNV in ctDNA, combined with the ctDNA level, can predict the prognosis of patients with HCC.

This study aimed to identify novel long noncoding RNA (lncRNA) biomarkers for hepatocellular carcinoma (HCC) using publicly available tissue genomic datasets and validate their diagnostic utility for early-stage HCC. Differentially expressed lncRNAs between 371 HCC and 50 nontumor tissues were obtained from The Cancer Genome Atlas liver hepatocellular carcinoma (TCGA&#95;LIHC) project. Subsequently, the expression of the serum- and extracellular vesicle (EV)-derived lncRNA was assessed in 10 patients with HCC and 10 healthy controls using RT-qPCR. The candidate lncRNAs were validated in 90 HCC and 92 non-HCC (29 healthy control, 28 chronic hepatitis, 35 liver cirrhosis) patients. The sensitivity, specificity, and area under the receiver operating characteristic curve (AUC) were calculated for the candidate lncRNAs and the current HCC biomarker, alpha-fetoprotein (AFP). SFTA1P, HOTTIP, HAGLROS, LINC01419, HAGLR, CRNDE, and LINC00853 were markedly upregulated in HCC in TCGA&#95;LIHC dataset. Among them, LINC00853 has not been reported in relation to HCC before. In patients with HCC, only expression of small EV-derived LINC00853 (EV-LINC00853) was increased. EV-LINC00853 showed excellent discriminatory ability in the diagnosis of all-stage HCC (AUC = 0.934, 95% confidence interval = 0.887-0.966). Moreover, using a 14-fold increase and 20 ng·mL -1 as cutoffs for EV-LINC00853 expression and AFP level, respectively, EV-LINC00853 was found to have a sensitivity of 93.75% and specificity of 89.77%, while AFP showed only 9.38% sensitivity and 72.73% specificity for the diagnosis of early-stage HCC (mUICC stage I). EV-LINC00853 had a positivity of 97% and 67% in AFP-negative and AFP-positive early HCC, respectively. Serum EV-derived LINC00853 may be a novel potential diagnostic biomarker for early HCC, especially for AFP-negative HCC., (© 2020 The Authors. Published by FEBS Press and John Wiley & Sons Ltd.)

The risk of developing hepatocellular carcinoma (HCC) after hepatitis B e antigen seroclearance (ESC) remains unclear. We established and validated a new risk prediction model for HCC development after ESC in patients with chronic hepatitis B (CHB) receiving antiviral therapy (AVT). Between 2006 and 2016, 769 patients (training cohort) and 1,061 patients (validation cohort) with CHB who experienced ESC during AVT using entecavir (ETV) or tenofovir disoproxil fumarate (TDF) were recruited. In the multivariate analysis, male sex (hazard ratio [HR] = 2.092; 95% confidence interval [CI] = 1.152-3.800), cirrhosis (HR = 5.141; 95% CI = 2.367-11.167) and fibrosis-4 index (FIB-4) of >3.25 (HR = 2.070; 95% CI = 1.184-3.620) were the independent risk factors for HCC development (all P < .05). Accordingly, a novel HCC-ESC AVT model was developed (1x[sex: male = 1, female = 0] + 3x(cirrhosis = 1, noncirrhosis = 0) + 1x(FIB-4: >3.25 = 1, ≤3.25 = 0). The cumulative risk for HCC development was significantly different among the risk groups based on the HCC-ESC AVT category (0-1, 2-4 and 5 for the low-, intermediate- and high-risk groups, respectively) (overall P < .001, log-rank test). The area under the receiver operating characteristic curve (AUC) for predicting HCC development 3, 5 and 10 years after ESC was 0.791, 0.771 and 0.790, respectively (all P < .05). The predictive value of the HCC-ESC AVT model was similar in the validation cohort (AUC = 0.802, 0.774 and 0.776 at 3, 5 and 10 years, respectively; all P < .05). Hence, we have developed and validated a new HCC-ESC AVT model for HCC development, which includes male sex, cirrhosis and FIB-4 of >3.25 as constituent variables., (© 2020 John Wiley & Sons Ltd.)

Purpose: To investigate the added value of considering hypointensity on late portal venous phase (LPVP) images as washout for diagnosis of hepatocellular carcinoma (HCC) using gadoxetic acid-enhanced MRI (Gd-EOB-MRI) in patients with chronic liver disease (CLD)., Methods: This retrospective study comprised 97 patients at high risk for HCC who underwent Gd-EOB-MRI including unenhanced, multi-arterial phase, conventional portal venous phase (CPVP, 60 s), and LPVP (mean, 99.9 ± 9.1 s; range, 90-119 s) images. A total of 115 hepatic lesions were identified by histopathological or clinical diagnosis. Three independent radiologists assessed the MRI images by consensus. Diagnosis of HCC was made using criteria of arterial hyperenhancement and hypointensity relative to the surrounding liver parenchyma (1) on CPVP or (2) on CPVP and/or LPVP images. The generalized estimating equation was used to compare diagnostic performance for HCC between Criterion 1 and 2., Results: In 82 HCCs, the frequency of hypointensity differed significantly between the CPVP and LPVP images (64.6% [53/82] vs. 84.1% [69/82], P < 0.001). Among 33 non-HCCs, two cHCC-CCs showed additional hypointensity on LPVP than CPVP images (33.3% [11/33] vs. 39.4% [13/33], P = 0.500). Criterion 2 provided significantly greater sensitivity for diagnosing HCC than Criterion 1 (54.9% [45/82] vs. 74.4% [61/82], P < 0.001), with relatively little reduction in specificity (90.9% [30/33] vs. 84.8% [28/33], P = 0.145)., Conclusion: Additional use of LPVP hypointensity as washout could significantly improve sensitivity for HCC diagnosis when utilizing Gd-EOB-MRI in patients with CLD, without a significant decrease in specificity.

Currently, a reliable serum biomarker for hepatocellular carcinoma (HCC) has not been established, particularly for early-stage HCC (single tumor < 2 cm). We aimed to investigate diagnostic serum exosomal microRNA (exo-miR) panel for early-stage HCC. Driver oncogenic miR (onco-miR) candidates were selected by integrative analysis of miR expression profiles from three different RNA sequencing datasets of human HCC. Expressions of selected onco-miRs in serum exosome were measured using quantitative real-time PCR. Diagnostic performances of serum exo-miRs for HCC were evaluated in the test cohort (N = 24) and validation cohort (N = 144). Serum exo-miR panels were developed using a logistic regression model, and their diagnostic performance was evaluated. Six promising driver onco-miRs, including miR-25-3p, miR-140-3p, miR-423-3p, miR-1269a, miR-4661-5p, and miR-4746-5p, were identified by integrative analysis of three different RNA sequencing datasets. Among the six candidates, four serum exo-miRs (miR-25-3p, miR-1269a, miR-4661-5p, and miR-4746-5p) showed promising performance in the test cohort with area under the receiving operator curve (AUROC) >0.8. In our validation study, serum exo-miR-4661-5p could diagnose HCC in all stages (AUROC = 0.917), even in early stage (AUROC = 0.923), with a greater accuracy than other candidate serum exo-miRs and serum AFP. The panel composed of exo-miR-4661-5p and exo-miR-4746-5p was identified as the most accurate biomarker for early-stage HCC (AUROC = 0.947, 95% confidence interval = 0.889-0.980, sensitivity = 81.8%, and specificity = 91.7%). In conclusion, exo-miR-4661-5p-based serum panel is a promising diagnostic marker for early-stage HCC., (© 2020 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.)

Objectives: Magnetic resonance elastography (MRE) is a non-invasive tool for measuring liver stiffness (LS) with high diagnostic accuracy. This study investigated whether quantified LS by MRE could predict early recurrence of patients with hepatocellular carcinoma (HCC) within the Milan criteria., Methods: A prospectively collected cohort, which included the HCC patients who underwent MRE before treatment (an HCC-MRE cohort), was analyzed. In the HCC-MRE cohort, only patients under the Milan criteria, who underwent hepatic resection, radiofrequency ablation (RFA), or transarterial chemoembolization (TACE), were reviewed. We investigated whether LS assessed by MRE was an independent predictor of early recurrence using Cox regressions and Kaplan-Meier analyses., Results: A total of 192 HCC patients under the Milan criteria who underwent hepatic resection (n = 96), RFA (n = 23), or TACE (n = 73) were included. Higher LS ratings (kPa; hazard ratio [HR] = 1.12; 95% confidence interval [CI] = 1.01-1.25; p = 0.040) emerged as an independent risk factor for early tumor recurrence. In the subgroup analysis, higher LS ratings were associated with higher risks of early HCC recurrence in both the resection/RFA group (> 4.5 kPa; HR = 2.95; 95% CI = 1.26-6.94; p = 0.013) and the TACE group (> 6 kPa; HR = 2.94; 95% CI = 1.27-6.83; p = 0.012)., Conclusion: LS assessed by MRE was an independent predictor of early recurrence among HCC patients under the Milan criteria after achieving a complete response., Key Points: • Liver parenchymal stiffness measured by MRE predicts early recurrence of treated HCC under Milan criteria. • A liver stiffness > 5.5 kPa was associated with worse recurrence-free survival. • Patients with high pre-treatment LS may benefit from stringent follow-up.

Background/aims: This study aimed to compare the diagnostic performances of Liver Imaging Reporting and Data System (LI-RADS) 2018 and Korean Liver Cancer Association-National Cancer Center (KLCA-NCC) 2018 criteria on magnetic resonance imaging (MRI) for the noninvasive diagnosis of hepatocellular carcinoma (HCC) in high-risk patients., Methods: This retrospective study included 273 treatment-naïve patients (71 patients with extracellular contrast agent [ECA]-MRI and 202 patients with hepatobiliary agent [HBA]-MRI; 352 lesions including 263 HCCs) with high risk of HCC who underwent contrast-enhanced MRI between 2016 and 2017. Two readers evaluated all lesions according to the criteria of LI-RADS 2018 and KLCA-NCC 2018. The per-lesion diagnostic performances were compared using the generalized estimating equation method., Results: On ECA-MRI, the sensitivity and specificity of LI-RADS 2018 and KLCA-NCC 2018 were not significantly different (LR-5 vs. definite HCC: 75.8% vs. 69.4%, P=0.095 and 95.8% vs. 95.8%, P>0.999; LR-5/4 vs. definite/probable HCC: 87.1% vs.83.9%, P=0.313 and 87.5% vs. 91.7%, P=0.307). On HBA-MRI, definite HCC of KLCA-NCC 2018 showed significantly higher sensitivity (79.1% vs. 68.2%, P<0.001) than LR-5 of LI-RADS 2018 without a significant difference in specificity (93.9% vs. 95.4%, P=0.314). Definite/probable HCC of KLCA-NCC 2018 had higher specificity (92.3% vs. 80.0%, P=0.003) than LR-5/4 of LI-RADS 2018. The sensitivity was lower for definite/probable HCC than for LR-5/4 without statistical significance (85.6% vs. 88.1%, P=0.057)., Conclusion: On ECA-MRI, LI-RADS 2018 and KLCA-NCC 2018 showed comparable diagnostic performances. On HBA-MRI, definite HCC of KLCA-NCC 2018 provided better sensitivity than LR-5 category of LI-RADS 2018 without compromising the specificity, while definite/probable HCC of KLCA-NCC 2018 revealed higher specificity than LR-5/4 of LI-RADS 2018 for diagnosing HCC.

Background & Aims: The liver Imaging Reporting and Data System (LI-RADS) is a comprehensive system for standardizing liver imaging in patients at high risk for hepatocellular carcinoma (HCC). We performed a meta-analysis to determine the diagnostic performance of the LR-5 category for HCC and the pooled proportions of HCCs in each LI-RADS category using CT/MRI LI-RADS v2017., Methods: We searched multiple databases for original studies reporting on the diagnostic accuracy of CT/MRI LI-RADS v2017. Random-effects models were used to determine the summary estimates of the diagnostic performance of the LR-5 category and the pooled proportions of HCCs for each LI-RADS category. Risk of bias and concerns regarding applicability were evaluated with the Quality Assessment of Diagnostic Accuracy Studies-2 tool., Results: Fourteen studies (3 prospective studies and 11 retrospective studies) were included in the final analysis, consisting of 2056 patients, 2589 observations, and 1693 HCCs. The pooled per-observation sensitivity was 67% (95% confidence interval [CI], 62%-72%) with specificity of 92% (95% CI, 88%-95%) in the LR-5 category of CT/MRI LI-RADS v2017 for diagnosing HCC. The pooled proportions of HCCs were 0% (95% CI, 0%-0%) for LR-1, 4% (95% CI, 0%-8%) for LR-2, 34% (95% CI, 23%-44%) for LR-3, 67% (95% CI, 53%-81%) for LR-4, and 92% (95% CI, 87%-96%) for LR-5. The proportions of HCCs were significantly different among LI-RADS categories 1-5 (P = .034)., Conclusions: The LR-5 category of CT/MRI LI-RADS v2017 shows moderate sensitivity and high specificity for diagnosing HCC. Higher LI-RADS categories contained higher proportions of HCCs., (© 2020 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Oxidative stress resistance in cancer cells has contributed to multi-drug resistance, which poses a serious challenge to cancer therapy. To surmount this, combinatorial treatment involving anticancer drugs and histone deacetylase inhibitors (HDACi) have emerged as a chemotherapeutic option. Yet, HDACi's role in redox states of cancer cells still requires elucidation. In the present study, we hypothesized that HDACi sensitizes cancer cells to oxidative stress and results in G2/M cell cycle arrest. Cell viability and cell cycle were analyzed using Cell Counting Kit 8 (CCK8) and fluorescent activated cell sorting (FACS), respectively. The transcriptomes of cells were investigated by massive analysis of cDNA end (MACE). Expression of mRNA and proteins were analyzed by quantitative real-time PCR (qPCR) and Western blot, respectively. Intracellular oxidative stress induced by tert-Butyl hydroperoxide (tBHP) reduced cell viability and resulted in G2/M cell cycle arrest in a dose-dependent manner in hepatocellular carcinoma (HCC) cells. The effects of sorafenib on cell cycle arrest and HCC viability were enhanced through HDACi treatment. MACE revealed that genes related to progression of G2/M cell cycle including Foxm1, Aurka, Plk1, and Ccnb1 were significantly down-regulated in tBHP and HDACi-treated HepG2 cells. Inhibition of FOXM1 with thiostrepton also resulted in reduced cell viability and expression of FOXM1 target genes such as Aurka, Plk1, and Ccnb1. These results indicate that HDACi sensitizes HepG2 cells to oxidative stress and results in G2/M cell cycle arrest via down-regulation of FOXM1, which plays a key role in progression of G2/M cell cycle., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Purpose: We compared the diagnostic performances of the European Association for the Study of the Liver (EASL) 2018 and Liver Imaging Reporting and Data System (LI-RADS) 2018 criteria on magnetic resonance imaging (MRI) for the noninvasive diagnosis of hepatocellular carcinoma (HCC) in high-risk patients and evaluated the difference in diagnostic value between MRI with extracellular contrast agents (ECA-MRI) and MRI with hepatobiliary agents (HBA-MRI)., Methods: This study included 382 observations from 298 patients at high risk for HCC who underwent preoperative multiphasic contrast-enhanced MRI between January 2015 and December 2016. Two readers assessed all observations according to the EASL 2018 and LI-RADS 2018 criteria, and the per-observation diagnostic performances were compared., Results: On ECA-MRI, the LR-5 category of LI-RADS 2018 showed significantly higher sensitivity (78.9% vs. 71.5%, p = 0.005) and accuracy (81.7% vs. 75.0%, p = 0.003) for the diagnosis of HCC than the EASL 2018. On HBA-MRI, the diagnostic performances of the EASL 2018 and LR-5 of LI-RADS 2018 were not significantly different. When using EASL 2018, no statistically significant differences were observed in the diagnostic performances between ECA-MRI and HBA-MRI; however, when using the LR-5 of LI-RADS 2018, ECA-MRI had a higher sensitivity (78.9% vs. 67.5%, p = 0.029) than HBA-MRI., Conclusions: On ECA-MRI, the LR-5 category of LI-RADS 2018 provides better sensitivity and accuracy than the EASL 2018 for diagnosing HCC. EASL 2018 provides comparable diagnostic performances between ECA-MRI and HBA-MRI, but the LR-5 category of LI-RADS 2018 provides better sensitivity on ECA-MRI than on HBA-MRI.

Background: Hypervascular hyperplastic nodules (HHNs) occasionally develop in patients with alcoholic liver cirrhosis (ALC) and show arterial enhancement, thus mimicking hepatocellular carcinoma (HCC). Importantly, HHN as a benign lesion should be distinguished from HCC., Purpose: To evaluate the value of gadoxetic acid-enhanced MRI (Gd-EOB-MRI) and diffusion-weighted imaging (DWI) in distinguishing HHN from small (<3 cm) hypervascular HCC (hHCC) in patients with ALC., Study Type: Retrospective case-control study., Field Strength/sequence: 3.0T/in- and out-of-phase, T 1 -weighted, T 2 -weighted, diffusion-weighted, apparent diffusion coefficient, and dynamic gadoxetic acid-enhanced images., Population: Among 560 patients with ALC who underwent Gd-EOB-MRI and DWI, 12 patients with 28 HHNs and 22 patients with 29 hHCCs smaller than 3 cm were included., Assessment: The following MRI features were evaluated by three independent radiologists: signal intensity (SI) on T 1 -weighted, T 2 -weighted, diffusion-weighted, and hepatobiliary phase (HBP) images; shape, homogeneity, and margin on HBP; diffusion restriction; intralesional fat; necrosis; hemorrhage; washout on portal venous phase (PVP) and/or transitional phase (TP); and capsular enhancement. Quantitative analysis was also conducted., Statistical Tests: Univariate and multivariate analyses were performed to determine the significant MRI findings, and their diagnostic performance for the prediction of HHN was analyzed., Results: Lesion size of ≤16 mm (odds ratio [OR], 24.41; P = 0.007), low-to-iso SI on DWI (OR, 26.92; P = 0.007), and absence of washout on PVP and/or TP (OR, 31.84; P = 0.009) were significant independent factors for predicting HHN. When all three criteria were satisfied, the specificity was 100%. Compared with hHCCs, HHNs showed significantly smaller size (mean, 13.8 mm vs. 19.9 mm; P < 0.001) and higher mean SI value (994.0 vs. 669.5) and lesion-to-liver SI ratio (1.045 vs. 0.806) on HBP (P < 0.001, respectively)., Data Conclusion: Gd-EOB-MRI and DWI may be helpful in differentiating HHN from small hHCC in patients with ALC., Level of Evidence: 3 Technical Efficacy Stage: 2 J. Magn. Reson. Imaging 2020;51:70-80., (© 2019 International Society for Magnetic Resonance in Medicine.)

Fibrolamellar hepatocellular carcinoma (FL‑HCC) is a variant of hepatocellular carcinoma (HCC) that most commonly affects adolescents and young adults and is associated with an extremely poor prognosis due to the lack of effective chemotherapeutic agents. Mutations in p53 are a common oncogenic driver in HCC but not in FL‑HCC. However, in tumors lacking a p53 mutation, the tumor suppressor activity of p53 has been revealed to be dysregulated in several different cancer types. One mechanism has been attributed to the overexpression of mouse double minute 4 protein (MDM4), a negative regulator of p53, which inhibits the normal functions of p53 including induction of apoptosis and DNA repair. Therefore, restoring the normal function of p53 in cancer cells by targeting MDM4 has become a potential therapeutic strategy. Hence, in the present study the components of the DNA damage response (DDR) pathway were examined; ATM, p53, and MDM4 in FL‑HCC. Seven FL‑HCC tumors along with their adjacent non‑neoplastic hepatic tissues were examined. Ataxia‑telangiectasia mutated (ATM), p53, and MDM4 protein expression was assessed using western blot analysis and cellular localization was determined using immunohistochemistry (IHC). MDM4 mRNA transcript levels were assessed using RT‑qPCR. The present results demonstrated that the DNA damage sensor, ATM, is phosphorylated and localized to the nuclei of tumor cells. While there was a significant increase in total p53 protein in tumor cells, phosphorylated p53 was revealed to preferably localize to the cytoplasmic compartment of tumor cells. Notably, the present results revealed that MDM4 transcript levels were increased in the majority of tumor samples and the nuclear MDM4 levels were significantly increased in tumor tissue compared to their adjacent non‑neoplastic liver tissue. The present results indicated that increased MDM4 expression and nuclear localization may be a potential mechanism for p53 dysregulation in FL‑HCC.

Background/aims: Barcelona Clinic Liver Cancer (BCLC) C stage demonstrates considerable heterogeneity because it includes patients with either symptomatic tumors (performance status [PS], 1-2) or with an invasive tumoral pattern reflected by the presence of vascular invasion (VI) or extrahepatic spread (EHS). This study aimed to derive a more relevant staging system by modification of the BCLC system considering the prognostic implication of PS., Methods: A total of 7,501 subjects who were registered in the Korean multicenter hepatocellular carcinoma (HCC) registry database from 2008 to 2013 were analyzed. The relative goodness-of-fit between staging systems was compared using the Akaike information criterion (AIC) and integrated area under the curve (IAUC). Three modified BCLC (m-BCLC) systems (#1, #2, and #3) were devised by reducing the role of PS., Results: As a result, the BCLC C stage, which includes patients with PS 1-2 without VI/EHS, was reassigned to stage 0, A, or B according to their tumor burden in the m-BCLC #2 model. This model was identified as the most explanatory and desirable model for HCC staging by demonstrating the smallest AIC (AIC=70,088.01) and the largest IAUC (IAUC=0.722), while the original BCLC showed the largest AIC (AIC=70,697.17) and the smallest IAUC (IAUC=0.705). The m-BCLC #2 stage C was further subclassified into C1, C2, C3, and C4 according to the Child-Pugh score, PS, presence of EHS, and tumor extent. The C1 to C4 subgroups showed significantly different overall survival distribution between groups (p<0.001)., Conclusions: An accurate and relevant staging system for patients with HCC was derived though modification of the BCLC system based on PS.

Circulating microbial dysbiosis is associated with chronic liver disease including nonalcoholic steatohepatitis and alcoholic liver disease. In this study, we evaluated whether disease-specific alterations of circulating microbiome are present in patients with cirrhosis and hepatocellular carcinoma (HCC), and their potential as diagnostic biomarkers for HCC. We performed cross-sectional metagenomic analyses of serum samples from 79 patients with HCC, 83 with cirrhosis, and 201 matching healthy controls, and validated the results in the same number of subjects. Serum bacterial DNA was analyzed using high-throughput pyrosequencing after amplification of the V3-V4 hypervariable regions of 16S rDNA. Blood microbial diversity was significantly reduced in HCC, compared with cirrhosis and control. There were significant differences in the relative abundances of several bacterial taxa that correlate with the presence of HCC, thus defining a specific blood microbiome-derived metagenomic signature of HCC. We identified 5 microbial gene markers-based model which distinguished HCC from controls with an area under the receiver-operating curve (AUC) of 0.879 and a balanced accuracy of 81.6%. In the validation, this model accurately distinguished HCC with an AUC of 0.875 and an accuracy of 79.8%. In conclusion, circulating microbiome-based signatures may be potential biomarkers for the detection HCC.

Background/aim: Fibroblast growth factor (FGF), vascular endothelial growth factor, and hepatocyte growth factor play a critical role in the pathogenesis of hepatocellular carcinoma (HCC)., Materials and Methods: We assessed nine single nucleotide polymorphisms (SNPs) in the FGF1, FGF2, FGF receptor (FGFR)-2, Flt-1, and c-MET genes in 245 HCC patients and 483 chronic hepatitis B virus (HBV) carriers without HCC., Results: Kaplan-Meier analysis showed that patients with the FGF2 rs308447 TT genotype had shorter overall survival than patients with the CC or CT genotype (p=0.016) and that FGF2 rs308379 A allele carriers had shorter overall survival than patients with the TT genotype (p=0.020)., Conclusion: Multivariate Cox proportional analysis revealed that the FGF2 rs308379 A allele (hazard ratio(HR)=1.663, p=0.004) and advanced tumor stage (HR=3.430, p<0.001) were independent prognostic factors for overall survival in patients with HCC., (Copyright© 2019, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Objectives: To compare the sensitivity of Liver Imaging Reporting and Data System (LI-RADS) v2017 for diagnosis of hepatocellular carcinoma (HCC) using multiphasic computed tomography (CT) between patients with and without moderate to severe fatty liver (MSFL)., Methods: This retrospective study included a total of 106 high-risk patients with 112 pathologically proven HCCs who underwent multiphasic CT. Patients were classified as MSFL (24 men, 2 women; mean age, 59.5 years [range, 38-79 years]) and non-MSFL (64 men, 16 women; mean age, 62.9 years [range, 40-89 years]) groups according to unenhanced CT liver and spleen parenchymal attenuation. Two independent radiologists assigned LI-RADS categories and accessed HCC features on CT. Sensitivities for LR-5 assignment and frequencies of HCC features were compared between the two groups., Results: Sensitivities of LR-5 assignment for diagnosing HCCs were not significantly different between MSFL and non-MSFL groups (65.4% [17/26] vs. 76.7% [66/86] for reviewer 1, p = 0.247; 73.1% [19/26] vs. 76.74% [66/86] for reviewer 2, p = 0.702). No significant differences in the frequencies of arterial hyperenhancement, washout, and capsule were observed between the two groups (96.2% [25/26] vs. 98.8% [85/86], p = 0.412; 80.8% [21/26] vs. 89.5% [77/86], p = 0.308; and 53.8% [14/26] vs. 57% [49/86], p = 0.778, respectively)., Conclusions: LI-RADS v2017 using CT showed comparable sensitivity for diagnosing HCC regardless of MSFL., Key Points: • Using LI-RADS v2017 with CT, diagnosis of HCC in patients with MSFL showed similar sensitivity to that in patients without MSFL. • Frequencies of major HCC features (arterial hyperenhancement, washout, and capsule) on CT between the MSFL and non-MSFL groups were not significantly different. • LI-RADS using CT may be feasible for diagnosing HCC in patients with fatty liver.

This study aimed to evaluate hepatic toxicity after repeated stereotactic body radiation therapy (SBRT) for recurrent hepatocellular carcinoma (HCC) using deformable image registration (DIR). Between January 2007 and December 2015, 85 patients who underwent two sessions of SBRT for HCC treatment were retrospectively analyzed. A DIR technique was used to calculate the cumulative dose of the first and second SBRT to the normal liver by matching two computed tomography simulation images. The Dice similarity coefficient (DSC) index was calculated to evaluate DIR accuracy. Before the first and second SBRT, 6 (7.1%) and 12 (14.1%) patients were Child-Pugh class B, respectively. Median tumor size was 1.7 cm before both SBRT treatments. Mean DSC index value was 0.93, being >0.9 in 79 (92.9%) registrations. Median cumulative mean liver dose (MLD) was 9.3 Gy (interquartile range, 7.6-11.7). Radiation-induced liver disease developed in three patients, and two of them, with Child-Pugh class B, experienced irreversible liver function deterioration following the second SBRT. The DIR method provided reliable information regarding cumulative doses to the liver. In patients with Child-Pugh class A liver function, repeated SBRT for small recurrent HCC could be safely performed with acceptable hepatic toxicity.

Background/aim: Epithelial cell adhesion molecule (EpCAM) is expressed in hepatic progenitor cells and hepatocellular carcinoma (HCC), and is considered a marker of liver cancer stem cells., Materials and Methods: A total of 262 patients were enrolled who had undergone surgical resection for HCC, with immunohistochemical staining results for EpCAM. The immunohistochemical expression of EpCAM and other stemness-related markers was evaluated as prognosticators of tumor recurrence and survival in patients who underwent surgical resection for HCC., Results: A multivariate Cox regression analysis showed that tumor size [hazard ratio (HR)=2.26, p=0.005], intrahepatic metastasis (HR=2.31, p=0.011), and EpCAM positivity (HR=1.74, p=0.038) were associated with tumor recurrence. In a Kaplan-Meier survival analysis, patients with EpCAM-positive tumors had a significantly higher tumor recurrence rate and a reduced overall survival compared to those with EpCAM-negative tumors., Conclusion: Immunohistochemical expression of EpCAM was identified as a poor prognosticator of recurrence and survival after surgical resection in patients with HCC., (Copyright© 2018, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

We evaluated whether nucleos(t)ide analog (NA) influences the risk of non-hepatocellular carcinoma (non-HCC) malignancies in patients with chronic hepatitis B (CHB). A total of 9867 patients with CHB were followed up for ≥12 months for the occurrence of any type of malignancy between 1998 and 2013. Patients who received NA for ≥180 days were defined as the NA group. Propensity score matching produced the control (n = 2220) and NA groups (n = 2220) after adjustment for age, sex, and the presence of diabetes mellitus and liver cirrhosis. The National Health Insurance Service sample cohort dataset was used for external validation. Regarding non-HCC malignancies, only old age was an independent risk factor (>50 years; hazard ratio 3.17, 95% confidence interval 1.71-5.88, P < .001) in multivariate analysis. With regard to specific cancers such as thyroid, breast, lung, stomach, colorectal, pancreatobiliary, and hematologic malignancy, there was no difference of the incidence of each malignancy between the NA and control groups in both the hospital-based and external validation cohorts. NA treatment neither raises nor lowers the incidence of non-HCC malignancies in patients with CHB. Patients >50 years old are encouraged to undergo surveillance for malignancies similar to the general population.

Aim: To compare the long-term therapeutic outcomes of repeated radiofrequency ablation (RFA) with that of transarterial chemoembolisation (TACE) in patients with local tumour progression (LTP) after initial RFA treatment for hepatocellular carcinoma (HCC)., Materials and Methods: This retrospective study was approved by the institutional review board and the requirement for informed consent was waived. Between July 2006 and February 2012, 713 patients underwent RFA for single HCC as a first-line treatment. Fifty-eight patients who showed LTP as initial tumour recurrence post-RFA treatment were included. Patients were treated with either repeated RFA (n=33) or TACE (n=25). TACE was performed as an alternative therapeutic option when repeated RFA was not feasible based on the planning ultrasonography. Recurrence-free and overall survival rates were estimated using the Kaplan-Meier method. Prognostic factors for outcomes were evaluated using the Cox proportional hazards model., Results: Both groups did not show significant differences in terms of baseline characteristics, with the exception being the proportion of subphrenic tumours (p=0.031). The RFA and TACE groups did not differ significantly in their 5-year recurrence-free and overall survival rates (17% versus 10.7% and 72.7% versus 51.9%, respectively, with all p-values >0.05). In addition, multivariate analyses revealed that type of treatment was not associated with recurrence-free or overall survival in patients with post-RFA LTP., Conclusion: TACE is an effective treatment, comparable to repeated RFA, in patients with LTP after initial RFA when repeated RFA is not feasible., (Copyright © 2017 The Royal College of Radiologists. Published by Elsevier Ltd. All rights reserved.)