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In eukaryotes, the ubiquitin-proteasome system (UPS) and the autophagy-lysosome pathway (ALP) are essential for maintaining cellular proteostasis and associated with cancer progression. Our previous studies have demonstrated that phosphatase and tensin homolog (PTEN), one of the most frequently mutated genes in human cancers, limits proteasome abundance and determines chemosensitivity to proteasome inhibitors in cholangiocarcinoma (CCA). However, whether PTEN regulates the lysosome pathway remains unclear.We tested the effects of PTEN on lysosome biogenesis and exosome secretion using loss- and gain-of-function strategies in CCA cell lines. Using in vitro dephosphorylation assays, we explored the regulatory mechanism between PTEN and the key regulator of lysosome biogenesis, transcription factor EB (TFEB). Using the migration assays, invasion assays, and trans-splenic liver metastasis mouse models, we evaluated the function of PTEN deficiency, TFEB-mediated lysosome biogenesis, and exosome secretion on tumor metastasis. Moreover, we investigated the clinical significance of PTEN expression and exosome secretion by retrospective analysis.PTEN facilitated lysosome biogenesis and acidification through its protein phosphatase activity to dephosphorylate TFEB at Ser211. Notably, PTEN deficiency increased exosome secretion by reducing lysosome-mediated degradation of multi-vesicular bodies (MVBs), which further facilitated the proliferation and invasion of CCA. TFEB agonist curcumin analog C1 restrained the metastatic phenotype caused by PTEN deficiency in mouse models, and we highlighted the correlation between PTEN deficiency and exosome secretion in clinical cohorts.In CCA, PTEN deficiency impairs lysosome biogenesis to facilitate exosome secretion and cancer metastasis in a TFEB phosphorylation-dependent manner.

Protein oxidation during food processing causes changes in the balance of protein-molecular interactions and protein-water interactions, ultimately leading to protein denaturation, which leads to the loss of a range of functional properties. Therefore, how to control the oxidative modification of proteins during processing has been the focus of research.In this study, the intrinsic fluorescence value of the myofibrillar proteins (MP) decreased and the surface hydrophobicity value increased, indicating that the heat treatment caused a significant change in the conformation of the MP. With the increase of heating temperature, protein carbonyl content increased, total sulfhydryl content decreased, and protein secondary structure changed from α-helix to β-sheet, indicating that protein oxidation and aggregation occurred. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) result showed that heat treatment can lead to the degradation of proteins, especially myosin heavy chain, but actin had a certain thermal stability. A total of 733 proteins were identified by proteomics, and the protein oxidation caused by low temperature vacuum heating (LTVH) was determined to be mild oxidation dominated by malondialdehyde and 4-hydroxynonenal by oxidation site division.This study revealed the effect of LTVH treatment on the protein oxidation modification behavior of sturgeon meat, and explored the effect mechanism of LTVH treatment on the processing quality of sturgeon meat from the perspective of protein oxidation. This result of this study may provide a theoretical basis for the precise processing of aquatic products. This article is protected by copyright. All rights reserved.

Oncogene-derived metabolic reprogramming is important for anabolic growth of cancer cells, which is now considered to be not simply rely on glycolysis. Pentose phosphate pathway and tricarboxylic acid cycle also play pivotal roles in helping cancer cells to meet their anabolic and energy demands. The present work focused on gankyrin, a relatively specific oncogene in hepatocellular carcinoma (HCC), and its impact on glycolysis and mitochondrial homeostasis. Metabolomics, RNA-seq analysis, and subsequent conjoint analysis illustrated that gankyrin regulated the pentose phosphate pathway (PPP), tricarboxylic acid (TCA) cycle, and mitochondrial function and homeostasis, which play pivotal roles in tumor development. Mechanistically, gankyrin was found to modulate HCC metabolic reprogramming via TIGAR. Gankyrin positively regulated the transcription of TIGAR through Nrf2, which bound to the antioxidant response elements (AREs) in the promoter of TIGAR. Interestingly, TIGAR feedback regulated the transcription of Nrf2 and subsequently gankyrin by promoting nuclear importation of PGC1α. The loop between gankyrin, Nrf2, and TIGAR accelerated glucose metabolism toward the PPP and TCA cycle, which provided vital building blocks, such as NADPH, ATP, and ribose of tumor and further facilitated the progression of HCC.

Based on comprehensive analysis of tectonic and fault evolution, core, well logging, seismic, drilling, and production data, the reservoir space characteristic, distribution, origin of fault-karst carbonate reservoir in Yueman block of South Tahe area, Halahatang oilfield, Tarim Basin, were studied systematically. And the regular pattern of hydrocarbon accumulation and enrichment was analyzed systematically based on development practice of the reservoirs. The results show that fault-karst carbonate reservoirs are distributed in the form of “body by body” discontinuously, heterogeneously and irregularly, which are controlled by the development of faults. Three formation models of fault-karst carbonate reservoirs, namely, the models controlled by the main deep-large fault, the secondary fault and the secondary internal fault, are built. The hydrocarbon accumulation and enrichment of fault-karst carbonate reservoirs is controlled by the spatiotemporal matching relation between hydrocarbon generation period and fault activity, and the size and segmentation of fault. The study results can effectively guide the well deployment and help the efficient development of fault-karst carbonate reservoirs of South Tahe area, Halahatang oilfield. Key words: Tarim Basin, Halahatang oilfield, South Tahe area, Ordovician, fault-karst, carbonate reservoir, hydrocarbon accumulation, oil-gas enrichment

Background and aims Cancer cell survival depends on the balance between reactive oxygen species production and scavenging, which is regulated primarily by NRF2 during tumorigenesis. Here, we demonstrate that deletion of RBP5-mediating protein (RMP) in an autonomous mouse model of intrahepatic cholangiocarcinoma (ICC) delays tumor progression. Approach and results RMP-overexpressing tumor cells exhibited enhanced tolerance to oxidative stress and apoptosis. Mechanistically, RMP competes with NRF2 for binding to the Kelch domain of KEAP1 (Kelch-like ECH-associated protein 1) through the E**E motif, leading to decreased NRF2 degradation via ubiquitination, thus increasing NRF2 nuclear translocation and downstream transactivation of antioxidant genes. This RMP-KEAP1-NRF2 axis promotes ICC tumorigenesis, metastasis, and drug resistance. Consistent with these findings, the RMP level in human ICC is positively correlated with the protein level of NRF2 and is associated with poor prognosis. Conclusion These findings reveal that RMP is involved in the oxidative stress defense program and could be exploited for targeted cancer therapies.

Sturgeon is one of the most precious fish resources worldwide. Low temperature vacuum heating (LTVH) has been confirmed as a good way of maintaining food quality. However, there is a lack of in-depth studies assessing the impact of LTVH on lipid oxidation and flavor formation.The present study compared the effect of LTVH and traditional cooking on lipid oxidation and flavor of sturgeon fillets. In total, 13 fatty acids were detected, of which polyunsaturated fatty acids content was the highest (P 0.05). LTVH prevented the formation of conjugated diene and thiobarbituric acid reactive substances (P 0.05), as manifested by an increased signal intensity of free radicals of electron spin resonance. The characteristic peaks intensity of lipid by Raman at 970 cmOverall, the results of the present study support the view that LTVH is a healthier way of cooking. © 2022 Society of Chemical Industry.

The development of hypersonic vehicles puts forward new requirements for simulation capabilities of ground wind tunnel facilities. MHD acceleration has brought great interests as a means of enhancing gas speed and become an important breakthrough. Two-dimensional numerical simulation of electron MHD(EMHD) acceleration process in continuous Faraday channel is carried out using MHD five-equation model of based on low magnetic Reynolds number. The thermal, electromagnetic, flow characteristics and its interactions in the channel are analyzed. The results show that the distribution of potential in the channel is no longer symmetrical due to the existence of Lorentz force, resulting in Hall effect. The current density whose maximum value exceeds 4×104 A/m2, is concentrated at the edge of the electrode, which would become more and more obvious as the magnetic induction intensity increases. Influenced jointly by the Lorentz force and the Hall effect, the direction of current shifts to the right, which in turn causes the Lorentz force shift upward. The effects of electrical conductivity σ, magnetic induction intensity B and applied voltage U on the flow field are analyzed. As σ increases, a large amount of Joule heat which reaches 3.24×108 J at σ=0.1 S/m is generated in the channel, making the Mach number of airflow decrease gradually. As B increases, the electromagnetic effect is significantly enhanced, while the Joule heat is reduced, about 32% at B=6 T. Therefore, the airflow is effectively accelerated. The load factor of EMHD channel is affected by applied voltage and magnetic field jointly. The effect of the applied voltage on airflow is more significant than that of magnetic induction intensity at small B. But at large B, the influence of the magnetic induction intensity on airflow will dominate.

Glutamine-fructose-6-phosphate aminotransferase (GFPT) is a key factor in the hexosamine metabolism pathway. It regulates the downstream factor O-GlcNAc to change cell function and plays an important role in the metabolism and immune process of tissues and organs. However, the evolutionary relationship of GFPT family proteins in vertebrates has not been elucidated.To deduce and explore the evolution and function of vertebrate GFPT family.18 GFPT sequences were obtained from Homo sapiens (H. sapiens), Trachypithecus francoisi (T. francoisi), Mus musculus (M. musculus), Rattus norvegicus (R. norvegicus), Gallus gallus (G. gallus), Zootoca vivipara (Z. vivipara), Xenopus tropicalis (X. tropicalis), Danio rerio (D. rerio), Rhincodon typus (R. typus), Plasmodium relictum from National Center for Biotechnology Information (NCBI). The physical and chemical characteristics and molecular evolution of GFPT family proteins and nucleic acid sequences were analyzed by ClustalX2, Gene Doc, MEGA-X, SMART, Datamonkey, R etc. RESULTS: Based on the neighbor-joining (NJ) phylogenetic tree and evolution fingerprints, GFPT family members of vertebrates can be divided into two groups: the GFPT1 group and the GFPT2 group. Seven positive selection sites were identified by IFEL and integrated methods mixed effects model of evolution (MEME) and fixed effects likelihood (REL). Finally, we predicted 28 phosphorylation sites and 18 ubiquitousness sites in the human GFPT1 sequence, 10 phosphorylation sites, and five ubiquitousness sites in GFPT2. Gene ontology (GO) analyzes the protein molecules and KEGG signaling pathways of vertebrates interacting with GFPT family proteins.Our work confirmed that higher animals GFPT family may have differentiated GFPT1 and GFPT2, which meets their own functional needs. This knowledge answers the question what the origin and evolution of GFPT family in vertebrates and provided the basis for disease treatment and function research of GFPT protein.

Gallbladder cancer (GBC) is an aggressive malignancy of biliary tract with poor prognosis. Although several studies have shown the frequency of relevant genetic alterations, there are few genetic models or translational studies that really benefit for GBC treatment in the era of precision medicine. By targeted sequencing and immunohistochemistry staining, we identified that phosphate and tension homology deleted on chromosome ten (PTEN) was frequently altered in GBC specimens, and loss of PTEN expression was independently correlated with poor survival outcomes. Further drug screening assays revealed proteasome inhibitor bortezomib as a promising agent for GBC treatment, and knockdown of PTEN increased bortezomib efficacy both in vivo and in vitro. Therapeutic evaluation of patient derived xenografts (PDXs) strongly supported the utilization of bortezomib in PTEN deficient GBC. Mechanically, functional PTEN inhibited ARE-dependent transcriptional activity, the same machinery regulating the transcription of proteasome subunits, thus PTEN suppressed proteasome activity and bortezomib sensitivity. Through siRNA screening, we identified the ARE-related transcriptional suppressor BACH1 involved in PTEN-mediated proteasome inhibition and regulated by PTEN-AKT1 axis. In summary, our study indicates that proteasome activity represents a prime therapeutic target in PTEN-deficient GBC tumors, which is worthy of further clinical validation.

Because of the large-size caves and fractures, carbonate reservoirs are heterogeneous. Based on the characteristics of fractured-cavernous reservoir, seismic and geology data, an interpretation model for well test of natured vuggy reservoir was developed in consideration of the existence of large-scale cave near the wellbore. The equations for expressing the pressure at bottom hole were obtained by using the concept of Green’s function and the Laplace transformation technique. The analysis on the affecting factors of dynamic pressure at well bottom indicates that with the existence of discontinuous cavity-region in the formation, the pressure derivative curve will concave downward which is caused by physical property differences between inside and outside the cavity. The size of cavity mainly affects the amplitude of the pressure derivative reduction. The results of a real testing case interpretation have good consistency with geological features, which proved the mathematical model.

This study aimed to reveal the molecular mechanisms associated with off-flavor generation in sturgeon fillets treated by low temperature vacuum heating (LTVH). Label-free quantitative proteomics was used to identify 120 favor-related proteins, 27 proteins were screened as differentially expressed for bioinformatics analysis. 17 of KEGG pathways were identified. Particularly, proteins involved in proteasome and peroxisome were highly correlated with off-flavor formation. They were primarily implicated in the structures of proteins, including binding and proteasome pathways. The results indicated that the LTVH reduced the binding sites by down-regulating protease and superoxide dismutase expression. LTVH increased the myofibrillar protein and sulfhydryl content and decreased the total volatile basic nitrogen and thiobarbituric acid reactive substance, which confirmed that protein oxidation was related to off-flavor. This proteomics study provided new insights into the off-flavor of sturgeon with LTVH, and proposed potential link between biological processes and off-flavor formation.

Circular RNAs (circRNAs) are relatively recently identified noncoding RNAs that are ubiquitously expressed in human tissues and serve key functions in regulating gene expression. However, few studies have focused on human intervertebral disc degeneration (IDD) circRNAs, and the potential role of circRNAs in IDD has not been described in detail. In the present study, circRNA expression data was downloaded from the Gene Expression Omnibus database, and circRNAs in the human intervertebral disc were classified according to their length, indicating their uniform distribution of circRNAs of different lengths. Gene Ontology analysis was performed, which indicated that the differentially expressed circRNAs were mainly produced as a result of catalytic activity and from binding genes in the molecular function category, cell part genes in the cellular component category, and cellular and metabolic process genes in the biological process category. Classification analysis divided the circRNAs host genes into 16 classes; with nucleic acid binding genes ranked as the most common host gene type in IDD tissue. Pathway analysis indicated that >15 signaling pathways may serve different roles in IDD, and Wnt signaling, gonadotropin‑releasing hormone receptor and integrin signaling pathways may serve important roles. Using co‑expression analysis, 76 differentially expressed circRNAs and host gene pairs were identified, which were divided into four groups: CircRNAs and their host genes downregulated; circRNAs downregulated and host genes upregulated; circRNAs and their host genes upregulated; and circRNAs upregulated and host genes downregulated. Finally, hsa_circ_0008305 upregulation and hsa_circ_0041946 downregulation were validated in IDD using reverse transcription‑quantitative polymerase chain reaction. In conclusion, the findings of the present study may shed light on the potential roles of circRNAs in IDD and the possibility for their use in the diagnosis and clinical treatment of IDD in the future.

This study was carried out to explore the roles of circular RNAs (circRNAs) in nucleus pulposus (NP) tissues in intervertebral disc degeneration (IDD). Differentially expressed circRNAs in IDD and normal NP tissues were identified based on the results of microarray analysis. Bioinformatics techniques were employed to predict the direct interactions of selected circRNAs, microRNAs (miR), and mRNAs. CircRNA_104670 was selected as the target circRNA due to its large multiplier expression in IDD tissues. After luciferase reporter and EGFP/RFP reporter assays, we confirmed that circRNA_104670 directly bound to miR-17-3p, while MMP-2 was the direct target of miR-17-3p. The receiver-operating characteristic (ROC) curve showed that circRNA_104670 and miR-17-3p had good diagnostic significance for IDD (AUC circRNA_104670 = 0.96; AUC miRNA-17-3p = 0.91). A significant correlation was detected between the Pfirrmann grade and expression of circRNA_104670 (r = 0.63; p = 0.00) and miR-17-3p (r = −0.62; p = 0.00). Flow-cytometric analysis and the MTT assay showed that interfering with circRNA_104670 using small interfering RNA (siRNA) inhibited NP cell apoptosis (p, Spinal disease: A roundabout route to disc degeneration ‘RNA sponges’ may provoke lower back pain by soaking up regulatory RNAs that normally protect the protein infrastructure surrounding cells in intervertebral discs. Many people suffer from lower back pain arising from disc degeneration (IDD). A team led by Fei-Zou and Jian-Yuan Jiang at Fudan University, Shanghai, China set out to identify molecular mechanisms that might contribute to IDD. They focused on circular RNAs, non-protein coding RNAs that have been linked to a variety of diseases. The researchers learned that IDD is associated with strongly elevated expression of a circular RNA that acts as an ‘RNA sponge’, binding to and thereby inactivating other RNA molecules. This inactivation ultimately results in the excessive production of an enzyme that can damage the protein matrix that supports cells within spinal discs, potentially setting up the conditions for IDD.

Adjuvant transcatheter arterial chemoembolization (TACE) protects against hepatocellular carcinoma (HCC) and is associated with reduced disease recurrence and improved outcome after surgery. However, deterioration of liver function after TACE negatively impacts the patient prognosis and limits it use as an option to prolong survival. We analyzed two independent cohorts that included a total of 510 patients with HCC who had undergone tumor resection. Immunohistochemistry assay was used to measure RPB5-mediating protein (RMP) expression and assessed their association with recurrence rate and response to therapy with adjuvant TACE. In patients with HCC, the expression of RMP in tumor is associated with age, gender, tumor size, portal venous invasion, TNM stages, BCLC stages and overall survival. Among patients with high RMP expression, adjuvant TACE after resection was associated with early recurrence. Even in the patients with small tumor size (no more than 5 cm) or no venous invasion, RMP status is associated with response to adjuvant TACE. RMP status in tumors may be a useful marker in estimating prognosis in patients with HCC and in assisting in the selection of patients who are likely to benefit from adjuvant TACE to prevent relapse.

CKAP4, one kind of type II trans-membrane protein, plays an important role to maintain endoplasmic reticulum structure and inhibits the proliferation of bladder cancer cells by combining its ligand anti-proliferative factor (APF). However, the biological function of CKAP4 in the progression of liver cancer has not been clearly demonstrated. In the present study, we knocked down or overexpressed CKAP4 in hepatocellular carcinoma (HCC) cells and cell proliferation, invasion, and migration capacities were investigated by CCK-8 and transwell assays. In vivo tumor model in mice was used to evaluate the role of CKAP4 on growth and metastasis of HCC. The data documented that HCC cells with high CKAP4 levels were featured by low proliferation capability as well as low invasion potential. Interestingly, we found that CKAP4 suppressed the activation of epithelial growth factor receptor (EGFR) signaling, which may partly explain the role of CKAP4 in cell biological behavior of HCC. Further study revealed that CKAP4 could associate with EGFR at basal status and the complex was reduced upon EGF stimulation, leading to release EGFR into cytoplasm. Thus, we demonstrate the novel mechanism, for the first time, expression of CKAP4 regulates progression and metastasis of HCC and it may provide therapeutic values in this tumor.

A cationic organic flocculant epichlorohydrin-dimethylamine(EPI-DMA) was employed for the treatment of simulated leather-making process wastewater. The study aims at investigating the flocculation performance of EPI-DMA for the leather dyeing wastewater which contains fat-liquoring agent wastewater and retanning agent wastewater. The results show that decoloration efficiency of EPI-DMA for dyeing wastewater can reach around 90% with proper dosage. For containing retanning agent wastewater, EPI-DMA also has good flocculation performance, but for containing fat-liquoring agent wastewater, flocculation performance of EPI-DMA was poor.

In this study, Regenerated chrome tanning agent is prepared from chrome sludge by hydrogen peroxide oxidation. The regenerated chrome tanning agent was characterized by IR which reveals that organic acid and protein polypeptide combined with chrome have been mostly removed. A small part of organic acid and protein polypeptide, not oxidized, increases the masking effect of the product. The application results show that the shrinkage temperature (Ts) of wet blue leather can reach 100°C and the leather has good feeling of handle and fullness which shows it has same properties with commercial chrome tanning agent.

Stable leather superfine powder suspension was got by superfine powder of leather solid wastes, maleic anhydride(MAH) and acrylic acid(AA). The best MAH、AA modifying conditions with grafting ratio as index were discussed by orthogonal test according L9(34) factor levels, and the results show that modified productsⅠis got by the total amount of MAH account for 50% of the amount weight of leather superfine powder at 80°C for 3 hours in the solution of N,N-dimethyl formamide (DMF). The MAH modified products was filtered and washed with water to reserve, and then was further modified by AA and MA being designed orthogonal test according L9(34)factor levels for optimum conditions with grafting ratio as index. The best modification conditions were as follows: ammonium persulfate(APS) as the initiator, the mass of initiator accounted for 2% of total weight of monomers , the volume ratio of AA and MA was 5, and the reaction temperature was 90 °C for 3 hours in 12% aqueous solution. The monomer grafting percentage of the products was 37.8%. The product was characterized by infrared spectroscopy (IR) and Size analyzer.

Water-based anionic polycarbodiimide crosslinker which the storage stability is more than 6 months was synthesized by isophorone diisocyanate(IPDI), sodium hydroxyethyl sulfonate(SHES), 3-methyl-1-phenyl-2-phosopholene-1-oxide(MPPO). Polycarbodiimide crosslinker was characterized by Fourier transform infrared spectroscopy (FTIR) and was applied to aqueous polyurethane resin coating agents which containing carboxyl groups. The mechanical properties, the swelling ratio of the film in water, 0.05mol/L Sodium hydroxyl and butyl acetate and the dry and wet rub and abrasion resistance were tested. The infrared spectrogram shows the characteristic peak of -N=C=N- appeared at 2200～2100cm-1 and the characteristic peak of -SO3-appeared at 1105cm-1.The results of application indicated that: WAPC was applied to polyurethane resin, tensile strength of the film increased from 1.485MPa to 3.752 MPa. The swelling rate in water，0.05mol/L Sodium hydroxyl，butyl acetate decreased from 444.9% to 126.1%，181.9% to 92.5%，351.2% to 186.4% respectively; WPAC was added to top coating of coating agent, it was founded that the dry and wet rub resistance were both increased.

Yang et al. identify a feedback loop between gankyrin, an oncoprotein overexpressed in human hepatocellular carcinoma (HCC), and Nrf2. The positive feedback modulates a series of antioxidant enzymes that lower intracellular reactive oxygen species to confer protection from mitochondrial damage and cell death., Oxidative stress status has a key role in hepatocellular carcinoma (HCC) development and progression. Normally, reactive oxygen species (ROS) levels are tightly controlled by an inducible antioxidant program that responds to cellular stressors. How HCC cells respond to excessive oxidative stress remains elusive. Here, we identified a feedback loop between gankyrin, an oncoprotein overexpressed in human HCC, and Nrf2 maintaining the homeostasis in HCC cells. Mechanistically, gankyrin was found to interact with the Kelch domain of Keap1 and effectively competed with Nrf2 for Keap1 binding. Increased expression of gankyrin in HCC cells blocked the binding between Nrf2 and Keap1, inhibiting the degradation of Nrf2 by proteasome. Interestingly, accumulation and translocation of Nrf2 increased the transcription of gankyrin through binding to the ARE elements in the promoter of gankyrin. The positive feedback regulation involving gankyrin and Nrf2 modulates a series of antioxidant enzymes, thereby lowering intracellular ROS and conferring a steadier intracellular environment, which prevents mitochondrial damage and cell death induced by excessive oxidative stress. Our results indicate that gankyrin is a regulator of cellular redox homeostasis and provide a link between oxidative stress and the development of HCC.

The functions of cytoskeleton-associated membrane protein 4 (CKAP4), one kind of type II transmembrane protein, are associated with the palmitoyl acyltransferase DHHC2. The objective of the current study was to investigate CKAP4/DHHC2 expression and its prognostic significance in patients with hepatocellular carcinoma (HCC).Two independent cohorts of 416 patients with HCC were enrolled. All the patients included had defined clinicopathologic and follow-up data. Using real-time polymerase chain reaction and immunohistochemical assay, CKAP4 and DHHC2 expression were evaluated. The association between CKAP4/DHHC2 expression and HCC-specific disease-free survival and overall survival was analyzed by Kaplan-Meier curves, the log-rank test, and Multivariate Cox regression analyses.The data documented that CKAP4 expression was much higher in HCC tumor tissues compared with adjacent normal tissues and its expression was significantly correlated with tumor size, intrahepatic metastases, portal venous invasion, and Barcelona Clinic Liver Cancer stage of disease in 2 cohorts of patients. On survival analysis, patients with high CKAP4 expression appeared to have a favorable overall survival and a longer disease-free survival compared with those with low expression. DHHC2 expression was also examined in tissue microarray analysis by immunohistochemistry and the results demonstrated that 87.6% of the cases had low expression of DHHC2. Kaplan-Meier analysis indicated that a high level of DHHC2 expression predicted favorable overall survival and disease-free survival rates in both the training cohort and validation set. Furthermore, the combination of CKAP4 and DHHC2 was found to have a more powerful efficiency in prognosis prediction than either one alone.To the best of our knowledge, the current study is the first to demonstrate that the expression of CKAP4 and its palmitoyl acyltransferase DHHC2 correlates with disease progression and metastasis in patients with HCC and may provide prognostic and therapeutic value.

Hepatocellular carcinoma (HCC) is believed to arise from tumor-initiating cells (T-ICs), which are responsible for tumor relapse and metastases. Portal vein tumor thrombus (PVTT) is raised from HCC and strongly correlated to a poor prognosis. However, the mechanism underling the formation of PVTT is largely unknown. Herein, we provide evidence that RNA polymerase II subunit 5 (RPB5)-mediating protein (RMP) was progressively upregulated in PVTT and overexpressed RMP appeared to increase T-ICs self-renewal. Moreover, RMP promoted metastases of PVTT cells and HCC cells in vitro and in vivo. Knockdown of RMP attenuated T-ICs self-renewal and reversed epithelial-mesenchymal transition (EMT) in HCC and PVTT cells. The neutralizing assays suggested that interleukin-6 (IL-6) had an indispensable role in RMP regulating metastases and self-renewal of HCC cells. Furthermore, the transcription of IL-6 was verified to be modulated by RMP via interaction with p65 and RPB5, through which expanding the T-IC/cancer stem cell populations, as well as inducing EMT was promoted. These results suggested that RMP may promote PVTT formation by promoting IL-6 transcription. Thus, RMP serves as a potent factor contributed to develop PVTT and a promising therapeutic target for HCC patients.

Background: Asparagine synthetase (ASNS) is associated with drug resistance in leukaemia, and the function of this enzyme in the context of hepatocellular carcinoma (HCC) is not clear. In this study, the relationship between ASNS expression and clinical outcomes after surgical resection was investigated, and the therapeutic value of ASNS was also evaluated. Methods: The expression of ASNS was evaluated in HCC samples by real-time PCR and immunohistochemistry assays. The correlation between ASNS expression and clinicopathological features was investigated. Potential clinicopathological prognostic factors were examined by univariate and multivariate survival analysis. Asparagine synthetase was overexpressed and knocked down in HCC cell lines to assess the influence of the enzyme on cell proliferation, migration and tumourigenicity. L-asparaginase was used to treat HCC cells with high or low levels of ASNS in vitro and in vivo to examine the therapeutic efficacy. Results: The expression of ASNS was higher in HCC tumour tissues and was closely correlated with the serum AFP level, tumour size, microscopic vascular invasion, tumour encapsulation, TNM stage and BCLC stage. Patients with low ASNS expression levels had a poor prognosis with respect to overall survival (OS). The multivariate survival analysis indicated that ASNS is an independent prognostic factor for OS. Furthermore, functional studies demonstrated that ASNS significantly inhibits the proliferation, migration and tumourigenicity of HCC cells. The knockdown of ASNS markedly increased sensitivity to L-asparaginase, indicating that cells with different ASNS protein levels have different sensitivities to L-asparaginase. Conclusion: The expression of ASNS is an independent factor affecting the survival of HCC patients, and low ASNS expression in HCC was correlated with worse surgical outcomes. The ASNS may be a promising therapeutic target for the treatment of HCC.

Macrophages (Mψ) are the major component of infiltrating leukocytes in tumors and exhibit distinct phenotypes according to the microenvironment. We have recently found that signal regulatory protein α (SIRPα), the inhibitory molecule expressed on myeloid cells, plays a critical role in controlling innate immune activation. Here, we identify that SIRPα is down-regulated on monocytes/Mψ isolated from peritumoral areas of hepatocellular carcinoma (HCC) samples, while its level is moderately recovered in intratumor Mψ. In vitro assays demonstrate that SIRPα expression is significantly reduced on Mψ when cocultured with hepatoma cells. This reduction is partly due to the soluble factors in the tumor microenvironment. Knockdown (KD) of SIRPα prolongs activation of nuclear factor kappa B (NF-κB) and PI3K-Akt pathways as Mψ encounter tumor cells, leading to an increased capacity of Mψ for migration, survival, and proinflammatory cytokine production. Enhanced Stat3 and impaired Stat1 phosphorylation are also observed in tumor-exposed SIRPα-KD Mψ. Adoptive transfer with SIRPα-KD Mψ accelerates mouse hepatoma cells growth in vivo by remolding the inflammatory microenvironment and promoting angiogenesis. SIRPα accomplishes this partly through its sequestration of the signal transducer Src homology 2-containing phosphotyrosine phosphatase (SHP2) from IκB kinase β (IKKβ) and PI3K regulatory subunit p85 (PI3Kp85). Conclusion: These findings suggest that SIRPα functions as an important modulator of tumor-polarized Mψ in hepatoma, and the reduction of SIRPα is a novel strategy used by tumor cells to benefit their behavior. Therefore, SIRPα could be utilized as a potential target for HCC therapy. (Hepatology 2013;58:680–691)