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Background/Aims: Hepatitis B virus (HBV) infection is a major cause of cirrhosis and hepatocellular carcinoma. Therefore, we aimed to obtain further information on HBV pathogenesis, and to search for novel putative molecules for anti-HBV therapy. Methods: We utilized Isobaric Tags for Relative and Absolute Quantitation (iTRAQ) to identify the secretory proteins that are differentially expressed in the HBV DNA-transfected HepG2.2.15 cell line and its parental HepG2 cell line. Immunohistochemistry (IHC) was employed to assess the clinical relevance of the observations. Small interfering (si)RNA-based silencing transfection methods were carried out to study the function of ENPP2. Results: Totally, 133 unique proteins were identified as differentially expressed in HepG2.2.15 cell line compared with HepG2 cell line. Ectonucleotide pyrophosphatase/phosphodiesterase family member 2 precursor (ENPP2) is one of the most significantly up-regulated secretory proteins associated with HBV replication. This differential expression of ENPP2 was further validated by real-time quantitative RT-PCR, Western Blot and immunohistochemical analysis. To study the function of ENPP2, we knockdown ENPP2 expression in HepG2.2.15 cell line by RNA interference. ENPP2 silencing increased HBV replication approximately 2.3-fold by enhancing, via the type I IFN signaling pathway, HBV cccDNA (covalently closed circular DNA) translation into viral RNA. Moreover, attenuation of ENPP2 expression inhibited both the invasion and migration ability of hepatoma cells in vitro via interacting with the molecules in the tumor microenvironment. Conclusion: Our study demonstrates that ENPP2 may be a novel anti-HBV target and indicate that suppression of its expression may inhibit the invasion and migration ability of hepatoma cells. [ABSTRACT FROM AUTHOR]

Background and Aims Surgical resection ( SR) and transarterial chemoembolization ( TACE) have been commonly applied for patients with huge hepatocellular carcinoma ( HCC). However, optimal treatment has not been established. Methods Between 2000 and 2009, 267 patients with huge HCC (≥ 10 cm) underwent TACE and 84 underwent SR as the first treatment. Propensity score matching generated a matched cohort composed of 152 patients. We investigated overall survival and possible prognostic factors. Results At baseline, the surgery group showed a tendency to have solitary tumor (72.6% vs 39.3%, P < 0.001), less vessel invasion (29.8% vs 51.3%, P < 0.001), and unilobar tumor extent (77.4% vs 50.9%, P < 0.001) than TACE group. During median follow up of 10 months (range: 0-103), the surgery group showed higher 1-, 3-, and 5-year overall survival rates than TACE group (73.8%, 54.8%, and 39.8% vs 37.8%, 16.3%, and 9.7%, respectively, P < 0.001). In the propensity score-matched cohort, baseline characteristics did not differ between the two groups. Surgery group showed higher 1-, 2-, and 3-year overall survival rates than TACE group (69.7%, 58.6%, and 51.7% vs 40.2%, 33.9%, and 18.5%, respectively, P < 0.001) during median follow up of 14.5 months (range: 0-103). Multivariate analysis revealed that male ( HR 1.90; 95% CI, 1.01-3.58; P = 0.048), albumin ( HR 0.54; 95% CI, 0.34-0.85; P = 0.008), ascites ( HR 1.77; 95% CI, 1.02-3.08; P = 0.044), and SR ( HR 0.44; 95% CI, 0.28-0.70; P = 0.001) were the independent prognostic factors associated with survival. Conclusion Comparing survival after SR and TACE, we showed that SR would be associated with better outcomes than TACE as the first treatment of huge HCC. [ABSTRACT FROM AUTHOR]

Background/Aims Acute hepatic failure ( AHF) is one of the most serious complications of transcatheter arterial chemoembolization ( TACE). The aims of this study were to investigate risk factors of AHF after TACE and to establish a predictive model for AHF. Methods In the evaluation set, a total of 820 patients who underwent TACE as a first treatment for hepatocellular carcinoma were included. The demographic, laboratory, radiological and treatment-related factors were analysed to identify risk factors for AHF after TACE and a predictive model was established using the identified risk factors. In the validation set, a different cohort of 438 patients was included to validate the predictive model. Results The incidence of post- TACE AHF was 15.1% (124/820). Multivariate analysis revealed that presence of portal vein thrombosis, high aspartate aminotransferase, bilirubin, and log alpha-foetoprotein levels, and low albumin and sodium levels were independent risk factors. A mathematical model was established using these independent risk factors, and the area under the receiver operating characteristic curve of the model was 0.773 (95% confidence interval, 0.726-0.820). The cut-off value of 9 had a sensitivity of 78.2%, a specificity of 72.3%, a positive likelihood ratio of 2.82, a negative likelihood ratio of 0.30, a positive predictive value of 28.9% and a negative predictive value of 95.8%. Conclusions The risk factors of post- TACE AHF were presence of portal vein thrombosis, high aspartate aminotransferase, bilirubin, and alpha-foetoprotein levels, and low serum albumin and sodium levels. A mathematical model to predict post- TACE AHF was established. [ABSTRACT FROM AUTHOR]

To highlight the societal burden of HBV infection in South Korea, we estimated the annual societal costs of HBV-related diseases for the year 2005. For the economic costs of HBV-infection-related diseases estimate, baseline data was collected from the Health Insurance Review and Assessment Service (HIRA) database. To complement data from the HIRA database, hospital charts from sample hospitals was reviewed and patient surveys were conducted. In 2005, the societal cost of HBV infection was 1.937 trillion KRW, including 474,642 million KRW of direct costs and 1.463 trillion KRW of indirect costs. The cost breakdown by disease was CHB at 465,167 million KRW, cirrhosis at 533,449 million KRW, hepatocellular carcinoma at 863,940 million KRW, and liver transplantation at 74,635 million KRW. The estimated amount is equivalent to 0.24% of the 2005 Korean GDP. This analysis emphasizes how important the prevention and treatment of these diseases are from the perspectives of the Korean society. [ABSTRACT FROM AUTHOR]

The aim of this study was to screen and identify novel B cell epitopes within the human heparanase protein and to investigate the impact of self-developed anti-heparanase polypeptide antibodies on growth and invasion of HCCLM6 human hepatocellular carcinoma cells in vitro. The flexible regions of secondary structure and the B cell epitopes of the human heparanase amino acid sequence were predicted by DNAStar and Bcepred software.The multiple antigenic peptides (MAP) of the epitopes were synthesized in eight-branched form. Rabbits were immunized with the eight-branched MAPs mixed with the universal T-helper epitope human IL-1β peptide (VQGEESNDK, amino acid 163–171). The immunogenicity of the synthesized peptides was evaluated by ELISA, western blot and immunohistochemistry. The impact of the self-developed rabbit anti-heparanase polyclonal antibodies on growth and invasion ability of HCCMLM6 cells were analyzed in a cell culture model. The cells were first treated with one of the three antibodies, respectively, and then measured by using MTT, flow cytometry, plate clone formation, invasion assay and heparan sulfate degrading enzyme assay. The three amino acid sequences 1–15 (MAP1), 279–293 (MAP2), and 175–189 (MAP3) in the large subunit of the human heparanase protein were predicted as its most potential epitopes. ELISA, western blot and immunohistochemistry analysis showed that all three MAPs were capable to induce high titer of serum antibodies. Antibodies induced by MAP1 and MAP2 were high specific. Furthermore, anti-MAP2 antibodies showed the strongest avidity towards liver cancer tissues. Under the treatment with the three anti-heparanase antibodies, respectively, the growth, cell cycle and clone formation of the cells remained unchanged when compared with a treatment with normal rabbit IgG. However, an inhibition of cell invasiveness and heparanase activity could be detected under the treatment with anti-MAP1- or anti-MAP2-antibody (with a terminal concentration of 100 μg/ml). The cell invasiveness was decreased by 54 and 38%, respectively, the heparanase activity by 43 and 39%, respectively. The multiple antigenic peptides MAP1 (AC 1–15) and MAP2 (AC 279–293) may be the dominant B cell epitopes in the human heparanase protein. The induced polypeptide antibodies can effectively inhibit the heparanase activity of HCCLM6 liver cancer cells and therefore influence their invasion ability, which provides a theoretic basis for the development of anti-heparanase antibodies and their clinical use as vaccine. [ABSTRACT FROM AUTHOR]

23-Hydroxybetulinic acid (23-HBA) is the efficient antitumor compound extracted from the roots of a Chinese Medicinal Herb, Pulsatilla chinensis (Bge) Regel. To evaluate the effect of radioiodination on cytotoxicity, 23-HBA was radioiodinated with 125I. 125I-23-HBA could be prepared in high yields and good radiochemical purity and was characterized using reverse phase HPLC. In ICR mice bearing Liver Cancer HepA tumor, 125I-23-HBA showed a tumor uptake of 2.1% ID/g at 2 h p.i. and 0.15% ID/g at 48 h p.i on i.v. injection. When injected intratumorally, greater tumor uptake and retention was observed (20% ID/g at 2 h p.i. and 4.6% ID/g at 48 h p.i. respectively). [ABSTRACT FROM AUTHOR]

Background: Dysregulated endocytosis of membrane proteins contributes significantly to several hallmarks of cancer. Basigin can enhance cancer progression, but its precise mechanism remains unclear. CD98 promotes cell spreading and tumorigenicity by triggering integrin clustering and enhancing cell adhesion to the extracellular matrix. The endocytosis and recyle of basigin and CD98 might play critical roles in cancer. Methods: The role of CD98 was confirmed in liver cancer cells by cell spreading in vitro and tumorigenicity by nude mice xenograft tumor assay in vivo; membrane expression of basigin and CD98 in SMMC-7721 was measured by FCAS; pull down and SPR analysis were uses to reveal the direct association between basigin and CD98; DsRed1 tagged CD98 was blocked in the cytoplasm in K7721 (whose basigin was knockn out) and had a well colocalization with ER and Rab5a positive recycling endosomes under co-focal; finally, by FRET imaging and FCAS we observed the internalization of basigin and CD98 was flotillin-1-regulated, and their recycle at early steps was Arf6-mediated. Results: Basigin and CD98 were highly expressed and co-localized on the human hepatocellular carcinoma (HCC) cell membrane; basigin can directly bind to CD98, mediating CD98 redistribution on the HCC cell membrane and activating the downstream integrin signaling pathway. Internalization of basigin and CD98 was flotillin-1 regulated the and their recycling was mediated by Arf6. This recycling process for basigin and CD98 promotes cell spreading and tumor growth in liver cancer xenografts. Conclusion: Basigin, as a redistribution chaperone of CD98, plays a critical role in promoting cell spreading and the progression of hepatocellular carcinoma. [ABSTRACT FROM AUTHOR]

The dielectric properties of human liver were determined by characterization of tissue absorption and coupling of electromagnetic energy in the electromagnetic field. In this study the ex-vivo dielectric properties of human hepatocellular carcinoma (well and moderately differentiated), liver hemangioma, hepatic fibrosis (stages S1 and S2), and normal liver tissue were measured and analyzed over the frequency range of 10 Hz to 100 MHz. The dielectric properties over the frequency range can reflect tissue information including biological macromolecules, vesicles, and cellular membrane; these information aids in distinguishing different physiological states and lesions. The ex-vivo conductivity, permittivity, resistivity, as well as the characteristic parameters between the lesions and normal liver were analyzed and their differences were also verified. The data can contribute to developing bioelectric applications for tissue diagnostics and creating more accurate computer models for medical applications. [ABSTRACT FROM AUTHOR]

Oral administration of β-carotene (BC) was found to exert opposite effects on plasma levels of vascular endothelial growth factor (VEGF) in two animal models. One study in nude mice injected via tail vein with hepatocarcinoma SK-Hep-1 cells showed that BC decreases the plasma VEGF level, whereas the other study in nude mice injected subcutaneously with prostate tumor PC-3 cells showed that BC increases the plasma VEGF level. Herein we investigated whether BC (0.5-20 μM) possesses diverse effects on VEGF secretion in SK-Hep-1, PC-3 and melanoma B16F10 cells. We found that incubation of SK-Hep-1 cells with BC (1-20 μM) for 6 h significantly decreased VEGF secretion, whereas BC (1-10 μM) significantly increased the VEGF secretion in PC-3 cells. However, these effects disappeared at 12 h of incubation. Similar effects occurred in VEGF mRNA and protein expression after treatment of SK-Hep-1 and PC-3 cells with BC for 6 h. In contrast, BC (0.5-20 μM) did not affect mRNA and protein expression and secretion of VEGF in B16F10 cells. We also found that the proliferation of SK-Hep-1 and B16F10 cells was significantly inhibited by 20 μM BC at 6 and 12 h of incubation, whereas the proliferation of PC-3 cells was significantly inhibited by 20 μM BC at 12 h of incubation. In summary, the present study demonstrated the tumor-specific effect of BC on VEGF secretion in different cancer cell lines. [ABSTRACT FROM AUTHOR]

Hepatocellular carcinoma (HCC) is currently the third most common cause of cancer-related death in the Asia-Pacific region. Our previous work showed that knockdown of CD98 significantly inhibits malignant HCC cell phenotypes in vitro and in vivo. The level of CD98 in the membrane is tightly regulated to mediate complex processes associated with cell-cell communication and intracellular signaling. In addition, the intracellular domain of CD98 (CD98-ICD) seems to be of vital importance for recycling CD98 to the membrane after it is endocytosed. The intracellular and transmembrane domains of CD98 associate with β-integrins (primarily β1 but also β3), and this association is essential for CD98 mediation of integrin-like signaling and complements dominant suppression of β1-integrin. We speculated that isolated CD98-ICD would similarly suppress β1-integrin activation and inhibit the malignant behaviors of cancer cells. In particular, the exact role of CD98-ICD has not been studied independently in HCC. In this study, we found that ectopic expression of CD98-ICD inhibited the malignant phenotypes of HCC cells, and the mechanism possibly involves β1-integrin suppression. Moreover, the expression levels of CD98, β1-integrin-A (the activated form of β1-integrin) and Ki-67 were significantly increased in HCC tissues relative to those of normal liver tissues. Therefore, our preliminary study indicates that ectopic CD98-ICD has an inhibitory role in the malignant development of HCC, and shows that CD98-ICD acts as a dominant negative mutant of CD98 that attenuates β1-integrin activation. CD98-ICD may emerge as a promising candidate for antitumor treatment. [ABSTRACT FROM AUTHOR]

Background: The survival following transarterial chemoembolization (TACE) alone is still low in unresectable hepatocellular carcinoma (HCC) with almost patients developing disease progression after treatment. There is need to investigate additional therapeutic options that would intensify the initial response to TACE. The present study was to retrospectively compare the outcome and evaluate the prognostic factors of stereotactic body radiation therapy (SBRT) alone or as an adjunct to transarterial embolization (TAE) or TACE in the treatment of HCC >5 cm. Methods: From January 2011 to April 2015, 77 patients received SBRT followed by TAE or TACE (TAE/TACE + SBRT group) and 50 patients received SBRT alone (SBRT group). The dose of SBRT was 30-50 Gy which was prescribed in 3-5 fractions. Eligibility criteria were: a longest tumor diameter >5.0 cm and Child-Turcotte-Pugh (CTP) Class A or B. Exclusion criteria included tumor thrombus, lymph node involvement and extrahepatic metastasis. Results: The median follow-up period was 20.5 months. Median tumor size was 8.5 cm (range, 5.1-21.0 cm). Median overall survival (OS) in the TAE/TACE + SBRT group was 42.0 months versus 21.0 months in the SBRT group. The 1-, 3- and 5-year OS was 75.5, 50.8, and 46.9 % in the TAE/TACE + SBRT group and was 62.4, 32.9, and 32.9 % in the SBRT group, respectively (P = 0.047). The 1-, 3- and 5-year distant metastasis-free survival (DMFS) was 66.3, 44.3, and 40.6 % in the TAE/TACE + SBRT group and was 56.8, 26.1, and 17.4 % in the SBRT group, respectively (P = 0.049). The progression-free survival (PFS) and local relapse-free survival (LRFS) were not significantly different between the two groups. In the entire patient population, a biologically effective dose (BED10) ⩾100 Gy and an equivalent dose in 2 Gy fractions (EQD2) ⩾74 Gy were significant prognostic factors for OS, PFS, LRFS and DMFS. Conclusions: SBRT combined with TAE/TACE may be an effective complementary treatment approach for HCC >5 cm in diameter. BED10 ⩾100 Gy and EQD2 ⩾74 Gy should receive more attention when the SBRT plan is designed. [ABSTRACT FROM AUTHOR]

Adiponectin blocks hepatocellular carcinoma (HCC) progression by inducing cell apoptosis through the modulation of C-Jun N-terminal kinase and mammalian target of rapamycin. However, the precise upstream signaling pathways or molecules remain elusive. In the present study, we analyzed the role of antioxidant protein thioredoxin (Trx) in adiponectin-induced apoptosis in HCC. Adiponectin treatment decreased the viabilities of both HepG2 and Huh7 HCC cells accompanied by increased accumulation of intracellular reactive oxygen species, as evidenced by 2',7'-dichlorodihydrofluorescein diacetate staining. Pretreatment of these cells with the deoxidant N-acetylcysteine blocked the inhibitory effect of adiponectin. Levels of Trx2 protein in both HCC cells were significantly decreased, and the level of Trx1 was significantly inhibited in Huh7 cells while unchanged in HepG2 cells. However, the redox state of Trx1 was altered from reduced to the oxidized form following adiponectin treatment in HepG2 cells. Overexpression of both Trx proteins rescued adiponectin-induced cell apoptosis, whereas mutated Trx proteins were less effective. Further analysis suggested that both ASK1 and JNK signaling are involved in this process. Trx1 and Trx2 proteins also manifested protective effects on HCC cells in response to adiponectin treatment in a xenograft tumor model. Furthermore, high levels of Trx proteins and low adiponectin expression levels were found in primary human HCC samples compared with paracancerous tissues. These results suggest that Trx proteins play important roles in mediating adiponectin-induced HCC cell apoptosis, thus providing new insights into the pathogenesis of HCC and identifying adiponectin and Trx proteins as potential combinational therapeutic targets for the treatment of HCC. [ABSTRACT FROM AUTHOR]

Background Sorafenib is an orally active multikinase inhibitor approved for the treatment of advanced hepatocellular carcinoma ( HCC). However, clinical parameters that may predict the treatment outcomes in sorafenib-treated advanced HCC patients remains unknown. Methods A total of 99 advanced ( BCLC C) HCC patients treated with sorafenib as an initial treatment modality from January 2007 to December 2011 were retrospectively reviewed. Overall survival was the primary endpoint for the analysis. Various clinical parameters including tumour stage and adverse effects to sorafenib were analysed. Univariate and multivariate analysis were carried out to identify clinical parameters predictive of the effect of sorafenib. Results There were 86 males and 13 females included in this study, with a median age of 53 years. The median overall survival was 91 days. Sixty-nine patients had Child-Pugh class A cirrhosis and 30 patients had Child-Pugh class B cirrhosis. Hepatitis B virus was the predominant cause of HCC (75.8%). Noted adverse effects were hand-foot syndrome, diarrhoea, fatigue, abdominal pain, nausea and stomatitis. The presence of hand-foot syndrome and diarrhoea and the absence of portal vein thrombosis and lymph node metastasis predicted a better overall survival in the multivariate analysis. Excluding the absence of lymph node metastasis, the same parameters were associated with a longer radiological time to progression. Conclusion Advanced HCC patients treated with sorafenib who experienced hand-foot syndrome and diarrhoea showed better overall survival than patients without these side effects. These side effects may be used as clinical parameters predictive of sorafenib response in patients with HCC. [ABSTRACT FROM AUTHOR]

HAb18G/CD147, a membrane spanning molecule and highly expressed in hepatocellular carcinoma (HCC)cells, was shown to stimulate the production of matrix metalloproteinases (MMPs) in the interaction of tumorcells and fibroblasts. Studies on the EMMPRIN/CD147 showed that CD147 extracellular domain is involvedin the induction of MMPs. To study the biological molecular function of HAb18G/CD147 extracellulardomain (HAb18G/CD147-ED) on production of MMPs following mediated tumor cell invasion, we isolatedfour novel monoclonal anibodies (MAbs)–1B3, 3B3, HAb18Gedomab1, and HAb18Gedomab2—againstHAb18G/CD147-ED by immunization of BALB/c mice with purified HAb18G/CD147-ED fragments, whichwere efficiently expressed in Escherichia coli. Gelatin zymography and Boyden chamber assays were used to identify the production of MMPs in the co-cultured human fibroblast and HCC cells, and to quantify themigrated cells in the presence of the generated MAbs. The results showed that two MAbs (1B3 and 3B3)inhibited the secretion of MMP-2 and the HCC cell invasion, whereas the other two MAbs (HAb18Gedomab1and HAb18Gedomab2) had a reverse function. FCM additive assay showed that four MAbs recognized differentepitopes of HAb18G/CD147-ED. Taken together, the results suggest that various regions ofHAb18G/CD147-ED participated in the regulation of MMP secretion. [ABSTRACT FROM AUTHOR]

The article discusses a study which characterized the basigin isoforms and the function of basigin-3 to inhibit hepatocellular carcinoma proliferation and invasion.