Process Of Hepatocellular Carcinoma Research Articles

Abstract 864: Pan-cancer analysis of lactic acid metabolism landscape across the Cancer Genome Atlas

Abstract Introduction: Metabolic reprogramming plays an important role in tumor occurrence and progression. Our studies confirmed the survival state of patients with different metabolic systems was heterogeneous and glycolytic metabolism may cause tumor progression. Based on these findings, lactic acid metabolism in glycolysis aroused our interest. This study evaluated the potential association between lactate metabolism gene and tumor clinical process in hepatocellular carcinoma, and proved that it can be used as an effective tool for survival prediction and treatment guidance of 32 kinds of tumor patients by establishing lactate score prognosis model. Methods: We used the expression profile, proteomics and epigenetics of HCC lactate gene set in the Cancer Genome Atlas Database to systematically describe the changes of lactate related genes. The differentially expressed genes were then verified in 32 clinical tumor data sets. A prognostic model of tumor lactic acid infiltration was constructed to evaluate its correlation with the prognosis of patients. Finally, we analyzed the effects of lame score, immunotherapy and drug response on the clinical process of tumor. Results: Firstly, we analyzed the differential expression of lactate metabolism genes in HCC and found that the prognosis of patients with high lactate infiltration was significantly lower than that of mild infiltration. AF angiogenesis factor and EMT factor in high infiltration may be the main reason for their differential expression. We constructed a tumor lactic acid prognosis model through six significantly different lactic acid genes, and found that lame score can be used as a predictor of immunotherapy. In addition, we proved the role of lame score in Pan cancer. PGA was positively correlated with lame score in most tumors. The prognosis of 16 tumors with high lame score group was significantly lower than that of low lame score group. There were significant differences in lame score between metastatic and non metastatic cancer groups of Cesc, kirc and kirp. The high and low risk groups of some tumors showed different dryness indices and dampness. At the same time, methylation gene modification has a significant impact on the Lame score of pan cancer, and the Lame score is used to link the drug tolerance and specific pathway enrichment of pan cancer tumor cell lines, confirming the targeted effect of Lame score on tumor treatment. Finally, we used the clinical characteristics and immunohistochemical analysis of a cohort study of real patients with breast cancer endocrine therapy (Tamoxifen) and hepatocellular carcinoma targeted therapy (Lenvartinib) to verify the clinical treatment prediction ability of lame score. Conclusions: Our study provides a comprehensive analysis of lactate metabolic across cancers and highlights the potential treatment of lactate targeted therapies for cancers. Citation Format: Xiaolin Wei, Lei Cai, Qian He. Pan-cancer analysis of lactic acid metabolism landscape across the Cancer Genome Atlas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 864.

The importance of BUD13 in the prognosis of hepatocellular carcinoma revealed by pan-cancer analysis.

RNA splicing is an essential process involved in many aspects of cell proliferation, survival and differentiation, and given the importance of RNA splicing in gene regulation, alterations in this biological behavior have been associated with many human cancers. BUD13 as an RNA binding protein (RBP) has been sparsely studied in tumors; consequently, there is a compelling need to further investigate the expression profile of BUD13 in human cancers to provide new molecular clues for the pathogenesis of hepatocellular carcinoma. For this porpuse, we used a series of bioinformatics methods to synthesize the relationship between BUD13 and prognosis, tumor microenvironment (TME), immune infiltration, tumor mutational load (TMB), and microsatellite instability (MSI), and tried to find the potential biological processes of BUD13 in tumors by GSEA and GSVA. And the association between the expression of BUD13 gene and prognosis was predicted by constructing a nomogram of hepatocellular carcinoma by multifactorial regression model. Results showed tgat in the present study, we found that elevated expression of BUD13 is associated with poorer OS in a numerous cancers, including ACC, KIRC, LGG, LIHC, READ, THYM, and UCS.More importantly, BUD13 expression levels were also significantly correlated with TME. Our results also indicated that BUD13 expression was closely associated with Pyroptosis genes and immune-related genes. Furthermore, BUD13 expression was associated with TMB, MSI and antitumor drug sensitivity in various cancer types. Functional bioinformatics analysis indicated that BUD13 may be involved in multiple signaling pathways and biological processes in hepatocellular carcinoma. Based on BUD13 expression, a risk factor model was found to predict OS in hepatocellular carcinoma. In conclusion, overall this study suggests that BUD13 expression is associated with poor prognosis and may be involved in the development and progression of hepatocellular carcinoma, which may be further explored as a potential prognostic marker and new targeted therapy.

Impurity Free and Scalable Process of Sorafenib Tosyalte: Hepatocellular Carcinoma

An efficient, impurities free and scalable process of hepatocellular carcinoma and renal cell carcinoma of sorafenib tosylate with good quality and higher yield.

Comprehensive analysis of the MIR4435-2HG/miR-1-3p/MMP9/miR-29-3p/DUXAP8 ceRNA network axis in hepatocellular carcinoma

A growing number of studies have shown that competitive endogenous RNA (ceRNA) regulatory networks might play important roles during the process of hepatocellular carcinoma (HCC). This study assessed the role of the ceRNA network in immune cell infiltration in HCC. Immune-related gene sets were downloaded from Molecular Signatures Database, and differentially expressed genes were screened based on TCGA HCC transcriptome data. The corresponding miRNAs with low expression and good prognostic implications, and the corresponding lncRNAs with high expression and poor prognostic were identified to construct ceRNA networks. The networks were utilized for clinical correlation analysis and risk model construction, and the CIBERSORT algorithm was applied to assess immune cell infiltration. In this study, the mRNA-miRNA-lncRNA model was used to construct a ceRNA network in HCC using immune-related differentially expressed mRNAs. Assessment of the MIR4435-2HG/hsa-miR-1-3p/MMP9/hsa-miR-29-3p/DUXAP8 ceRNA network axis in HCC showed that a high risk/poor prognosis was significantly correlated with tumor stage and invasion depth. MMP9 was positively correlated with resting M0 macrophages and NK cells and negatively correlated with activated mast cells, resting mast cells, monocytes and activated NK cells. DUXAP8 was positively correlated with M2 macrophages and negatively correlated with MIR4435-2HG, which was positively correlated with M2 macrophages and negatively correlated with activated mast cells, CD8 T cells and follicular helper T cells. The correlation of the MIR4435-2HG/hsa-miR-1-3p/MMP9/hsa-miR-29-3p/DUXAP8 ceRNA network axis with immune cell infiltration provides further information on the mechanism of HCC development. The result might improve our understanding the interactions between immune related genes and non-coding RNAs in the occurrence and development of HCC, and the relevant RNAs might be used as diagnostic and prognostic biomarkers and molecular targets in HCC patients.

LncRNA ILF3-AS1 promotes cell migration, invasion and EMT process in hepatocellular carcinoma via the miR-628–5p/MEIS2 axis to activate the Notch pathway

BackgroundLong non-coding RNAs (lncRNAs) are essential indicators for hepatocellular carcinoma. LncRNAs can exert the same functions as their antisense mRNAs. ILF3 is an oncogene in hepatocellular carcinoma. ILF3 divergent transcript (ILF3-AS1) is the antisense RNA of ILF3, and has been reported as an oncogene in various cancers. AimsTo explore the role of lncRNA ILF3-AS1 in malignant phenotypes of hepatocellular carcinoma cells. Methods and resultsRT-qPCR analysis revealed that ILF3-AS1 was significantly upregulated in hepatocellular carcinoma cells. The hepatocellular carcinoma cell viability was suppressed by silenced ILF3-AS1. Transwell and wound healing assays showed that ILF3-AS1 downregulation inhibited cell invasion and migration. The levels of proteins associated with epithelial-mesenchymal transition (EMT) process and the Notch pathway were detected by western blot analysis. Luciferase reporter, RNA pull down and RIP assays were used to investigate the relationship between ILF3-AS1 and downstream target genes. ILF3-AS1 competed with meis homeobox 2 (MEIS2) for miR-628–5p in hepatocellular carcinoma cells. ILF3-AS1 elevated the levels of key proteins on the Notch pathway. Rescue assays demonstrated that MEIS2 reversed the antitumor effects of silenced ILF3-AS1 on hepatocellular carcinoma. In vivo assays demonstrated that ILF3-AS1 silencing inhibited the hepatocellular carcinoma tumor growth. ConclusionsILF3-AS1 promoted hepatocellular carcinoma progression via the Notch pathway and miR-628–5p/MEIS2 axis.

Microenvironment characterization and multi-omics signatures related to prognosis and immunotherapy response of hepatocellular carcinoma

BackgroundImmune cell infiltration in the tumor microenvironment (TME) affects tumor initiation, patients’ prognosis and immunotherapy strategies. However, their roles and interactions with genomics and molecular processes in hepatocellular carcinoma (HCC) still have not been systematically evaluated.MethodsWe performed unsupervised clustering of total 1000 HCC samples including discovery and validation group from available public datasets. Immune heterogeneity of each subtype was explored by multi-dimension analysis. And a support vector machine (SVM) model based on multi-omics signatures was trained and tested. Finally, we performed immunohistochemistry to verify the immune role of signatures.ResultsWe defined three immune subtypes in HCC, with diverse clinical, molecular, and genomic characteristics. Cluster1 had worse prognosis, better anti-tumor characteristics and highest immune scores, but also accompanied by immunosuppression and T cell dysfunction. Meanwhile, a better anti-PD1/CTLA4 immunotherapeutic response was predicted in cluster1. Cluster2 was enriched in TAM-M2 and stromal cells, indicating immunosuppression. Cluster3, with better prognosis, had lowest CD8 T cell but highest immune resting cells. Further, based on genomic signatures, we developed an SVM classifier to identify the patient’s immunological status, which was divided into Type A and Type B, in which Type A had poorer prognosis, higher T cell dysfunction despite higher T cell infiltration, and had better immunotherapeutic response. At the same time, MMP9 may be a potential predictor of the immune characteristics and immunotherapeutic response in HCC.ConclusionsOur work demonstrated 3 immune clusters with different features. More importantly, multi-omics signatures, such as MMP9 was identified based on three clusters to help us recognize patients with different prognosis and responses to immunotherapy in HCC. This study could further reveal the immune status of HCC and provide potential predictors for immune checkpoint treatment response.

A novel pretreatment model identifying high-risk of refractoriness after transcatheter arterial chemoembolization in unresectable hepatocellular carcinoma.

e16641 Background: Refractoriness to transcatheter arterial chemoembolization is common during the therapeutic process of hepatocellular carcinoma, which is an intractable issue and may compromise the prognosis. We aim to establish a pre-treatment model to identify patients with high risks of refractoriness. Methods: From 2010 to 2016, 824 patients who had initially underwent at least two sessions of transcatheter arterial chemoembolization in Zhongshan Hospital, Fudan University were retrospectively enrolled. These patients were randomly allocated into a training cohort and a validation cohort. The pre-treatment scoring model was established based on the clinical and radiological variables using logistic regression and nomogram. The discrimination and calibration of the model were also evaluated. Results: Logistic regression identified vascularization pattern, ALBI grade, serum alpha-fetoprotein level, serum γ- glutamyl transpeptidase level and major tumor size as the key parameters related to refractoriness. The p-TACE model was established using these variables (risk score range: 0-19.5). Patients were divided into six risk subgroups based on their scores ( < 4, ≥4, ≥7, ≥10, ≥13, ≥16). The discriminative ability, as determined by the area under receiver operating characteristic curve was 0.784 (95% confidence interval: 0.741-0.827) in the training cohort and 0.743 (95% confidence interval: 0.696-0.789) in the validation cohort. Moreover, satisfactory calibration was confirmed by Hosmer-Lemeshow test with P values of 0.767 and 0.913 in the training cohort and validation cohort. Conclusions: This study presents a pre-treatment model to identify patients with high risks of refractoriness after transcatheter arterial chemoembolization and shed light on clinical decision making.

LncRNA LINC00668 promotes cell proliferation, migration, invasion ability and EMT process in hepatocellular carcinoma by targeting miR-532-5p/YY1 axis.

Liver cancer is now one of the most lethal and commonest cancers in the world, among which over 90% is hepatocellular carcinoma (HCC). Recent studies have confirmed long non-coding RNAs (lncRNAs) are implicated in carcinogenesis. It has been reported lncRNA LINC00668 serves as an oncogene in several cancers. However, the mechanism where LINC00668 regulates HCC is still unclear. qRT-PCR analysis was adopted to detect the expression of relative RNAs. Cytoplasmic and nuclear RNA fraction analysis was conducted to verify the underlying molecular mechanism. Cell colony formation was carried out to test cell colony formation ability and transwell assays were performed to testify cell migratory and invaded abilities. Relevant protein expression level was measured by Western blot assay. LINC00668 was significantly up-regulated in HCC tissues and cell lines. LINC00668 knockdown inhibited cell proliferative, migratory and invasion abilities and slowed down the epithelial–mesenchymal transition (EMT) process. Mechanistically, LINC00668 positively modulates the expression of YY1 by competitively binding to miR-532-5p. It was revealed that LINC00668 up-regulation accelerated cell proliferation and motility in HCC and suggested LINC00668 could be a potential therapeutic target for HCC.

CDK1-PLK1/SGOL2/ANLN pathway mediating abnormal cell division in cell cycle may be a critical process in hepatocellular carcinoma

ABSTRACT This study aims to investigate the potential mechanisms and identify core biomarkers of Hepatocellular carcinoma (HCC). The profile GSE113850 was downloaded to analyze the differentially expressed genes. Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and protein–protein interaction network analysis were used to reveal the main signal pathways of the differentially expressed genes (DEGs) and hub genes. The correlation between core gene expression and pathological stages, the disease-free survival analysis, the overall survival analysis were analyzed by Gene Expression Profiling Interactive Analysis. Furthermore, we reidentified the expression level of core genes of carcinoma tissues and para-carcinoma tissues from 14 HCC patients with real-time reverse transcription-polymerase chain reaction analysis (RT-PCR) and western blotting. After SK-Hep1 cell was treated with cyclin-dependent kinase 1 (CDK1) siRNA for 72 h, we detected the expression of the core genes and fluorescence-activated cell sorting analysis. A total of 378 DEGs were found. GO and KEGG analysis revealed that the DEGs were mainly enriched in the cell cycle. There were positive correlations among CDK1, polo-like kinase 1, shugoshin2 and anillin actin-binding protein. Moreover, the expression levels of four core genes were related to the HCC occurrence, pathological stages, and survivorship curve. The clinical HCC specimens verified the higher expression level of core genes by real-time RT-PCR. The transfection of siCDK1 in SK-Hep1 resulted in a disordered cell cycle. Furthermore, CDK1 knockdown suppressed the expression of PLK1, ANLN, and SGOL2. The CDK1-PLK1/SGOL2/ANLN pathway mediating abnormal cell division in the cell cycle might be a critical process in HCC.

Activated hepatic stellate cells promote angiogenesis in hepatocellular carcinoma by secreting angiopoietin-1.

Angiogenesis is the central pathological process in hepatocellular carcinoma (HCC), and its progression is affected by tumor cells and the microenvironment. Activated hepatic stellate cells (aHSCs) are the most significant stromal cells involved in HCC. This study was aimed to explore the effects and mechanisms of aHSCs on angiogenesis in HCC. We isolated primary hepatoma cells, aHSCs, and hepatic vascular endothelial cells from human HCC samples. Then, we performed a novel in vitro assay and in vivo experiment in a nude mouse HCC model to investigate the functions of aHSCs on angiogenesis in HCC. Our results demonstrated that aHSCs are the primary sources of angiopoietin-1 (Ang-1) in human HCC in vitro, and aHSCs could promote hepatic vascular endothelial cell (HVEC) growth by secreting Ang-1. Furthermore, aHSCs could induce HVEC microtubule formation, and this ability was reduced when Ang-1 expression was silenced in aHSCs. In addition, CD34 expression in a nude mouse HCC model was downregulated when Ang-1 messenger RNA was silenced in aHSCs. Our data also indicated that HSC Ang-1 expression could be inhibited by overexpressing Raf kinase inhibitor protein. Therefore, we suggest that aHSCs promote angiogenesis through secreting Ang-1, potentially providing an interesting target for antiangiogenic therapies for HCC.

7-Deoxynarciclasine shows promising antitumor efficacy by targeting Akt against hepatocellular carcinoma.

Akt is a promising therapeutic target for cancer treatment. In our study, we have identified that 7-deoxynarciclasine (7-DONCS) is a potential inhibitor of Akt, which results in the repression of multiple oncogenic processes in hepatocellular carcinoma (HCC). We have found that 7-DONCS suppresses the growth of HCC by inducing the apoptotic and autophagic capacities, as well as by inhibiting epithelial-mesenchymal transition (EMT) in vitro and in vivo. Pretreatment of cells with specific autophagy inhibitor (Bafilomycin A1) or knockdown of endogenous LC‐3B by siRNA strongly enhences 7‐DONCS‐regulated apoptosis and EMT. Consequently, we have found that 7-DONCS selectively inhibits phospho-Akt (Ser473), and subsequent molecular docking reveals that 7-DONCS directly binds to the C-terminal domain of Akt. Overexpressing Akt significantly blocks these effects via 7-DONCS in HCC cells. Furthermore, 7-DONCS, by targeting Akt, exhibits a promising therapeutic effect in orthotopic hepatocellular tumors. Finally, higher p-Akt expression is associated with poor prognosis, and higher level of Akt was positively correlated with the enrichment of both apoptosis and autophagy downregulation, and EMT upregulation in HCC patients. These studies suggest that 7-DONCS serves as an attractive drug candidate by targeting Akt for future HCC therapy.

Abstract 1902: Enrichment of putative cancer stem cells during anti-angiogenic therapies promotes relapse formation in hepatocellular carcinoma

Abstract Background and Aims: Activation of neo-angiogenic processes in hepatocellular carcinoma (HCC) during disease progression is frequently associated with poor clinical outcome. Consequently, inhibition of neo-angiogenesis is an effective treatment strategy for advanced HCC. However, development of chemoresistance is observed in the majority of patients. Compelling evidence suggest that cancer stem cells (CSCs) may contribute to the acquisition of resistant properties in many solid tumors, but their exact role in this process for HCC remains to be defined. Here, we evaluate the importance of CSCs in the development of resistance and relapse formation after exposure to different anti-angiogenic therapies in HCC and define the concomitant adaptive molecular changes. Methods: Five HCC cell lines and one primary HCC isolate were exposed to sorafenib and sunitinib for a total of 14 days. The treatment effects on CSCs were estimated by sphere forming capacity in vitro as well as the side-population (SP) approach. Expression levels of key oncogenic and CSC markers, such as EpCAM, CD133 and ABCG2 transporter, were assessed by qRT-PCR and flow cytometry. Furthermore, whole transcriptome analyses were performed at different time points. Results: Both treatment regimens effectively reduced oncogenic properties in all investigated HCC cells. However, sustained anti-proliferative effect after treatments was observed in only one cell line. In three other hepatoma lines an initial treatment effect was subsequently followed by rapid re-growth thereby mimicking the responses observed in patients. Interestingly, two cell lines showed differential response to applied drugs, showing anti-proliferative effects to sorafenib, while relapse formation occurred after sunitinib treatments. While anti-oncogenic effects in sensitive cell lines were associated with significant reduction in sphere forming capacity, CSC marker EpCAM as well as SP cells, resistant cell line showed a transient increased in CSC properties. Importantly, acquired resistance to both drugs uniformly developed in the cell lines suggesting that common molecular mechanisms might be operative. These adaptive molecular changes involved signaling pathways known to be associated to cell survival and proliferation (RAS, AKT, MYC), as well as angiogenesis (VEGFR, PDGFR, HIF1a). Furthermore, the resistant cell lines showed compensatory upregulation of key oncogenic molecules such as EGFR as well as multidrug resistance ABC transporters. Conclusions: Our in vitro model recapitulates features of drug resistance observed in human HCC patients. Resistance to anti-angiogenic therapies might be fueled by transient expansion of CSCs. Therefore, specific targeting of CSCs as well as pro-oncogenic compensatory signaling pathways might be an effective therapeutic strategy to overcome resistance in HCC. Citation Format: Darko Castven, Carolin Czauderna, Diana Becker, Marcus A. Wörns, Snorri S. Thorgeirsson, Peter Grimminger, Hauke Lang, Peter R. Galle, Jens U. Marquardt. Enrichment of putative cancer stem cells during anti-angiogenic therapies promotes relapse formation in hepatocellular carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1902. doi:10.1158/1538-7445.AM2017-1902

A single nucleotide variant in microRNA-1269a promotes the occurrence and process of hepatocellular carcinoma by targeting to oncogenes SPATS2L and LRP6

Hepatocellular carcinoma (HCC) is one of the malignant and lethal cancers. Single nucleotide polymorphisms (SNPs) in microRNAs(miRNAs) can affect the expression and target identification of miRNAs and lead to the formation of malignant tumors. However, little is known about whether microRNA-1269a (miR-1269a) SNPs affect the susceptibility and progression of HCC or their specific mechanism. The association between microRNA-1269a rs73239138 and the susceptibility to HCC was verified by MassARRAY assay in a large case-control sample. The effect of miR-1269a and the variant on the proliferation and apoptosis of HCC cells was examined by flow cytometry (FCM), CCK8 assay and Western blot. The target of miR-1269a was identified by bioinformatics analysis and qRT-PCR and its role on cell proliferative capacity was examined by CCK8 assay. The expression level of miR-1269a was analyzed by qRT-PCR in HCC cells transfected with wild or variant type pre-miR-1269a plasmid.MiR-1269a produced a tumor suppressor effect by inhibiting cell proliferation and inducing apoptosis of human HCC cells, possibly via inhibiting the expression of its target genes SPATS2L and LRP6, which were tumor promoters. While, rs73239138 (G>A) in miR-1269a reduced the anticancer effect of miR-1269a possibly by attenuating its total amount in HCC cells or its target recognition, reduce its inhibition on target genes and promoted the susceptibility to HCC. Our findings for the first time proved that miR-1269a SNP plays a role in the occurrence and process of HCC and the relevant mechanism, in accompany with the discovery of the novel target genes of miR-1269a.

Angiogenesis in hepatocellular carcinoma

The published data concerning angiogenesis processes in hepatocellular carcinoma are systematized in the present review. The basic morphogenetic steps of neovascularization, namely, sinusoidal capillarization, tissue arterialization, and vascular mimicry and cooperation, are considered. A detailed analysis of activators and inhibitors of angiogenesis in tumors in general, and hepatocellular carcinoma in particular, is presented. The key parameters of angiogenesis and growth factors associated with clinical characteristics and prognosis of the disease are enumerated. The progressive increase in blood supply through the hepatic artery and consequent reduction of blood flow through the portal vein system concomitantly to the increase of the degree of malignancy of hepatocellular carcinoma are emphasized. Special attention is paid to the development of approaches for the detection of markers for specific stages of angiogenesis and antiangiogenic therapies for malignant tumors.

FGF19 promotes epithelial-mesenchymal transition in hepatocellular carcinoma cells by modulating the GSK3β/β- catenin signaling cascade via FGFR4 activation.

Compelling evidence suggests that the epithelial-mesenchymal transition (EMT) correlates with aggressiveness of tumors and poor survival. FGF19 has been shown to be involved in EMT in cholangiocarcinoma and colorectal cancer, however, molecular mechanisms underlying FGF19-induced EMT process in hepatocellular carcinoma (HCC) remain largely unknown. Here, we show the expression of FGF19 is significantly elevated and negatively associated with the expression of E-cadherin in HCC tissues and cell lines. Ectopic FGF19 expression promotes EMT and invasion in epithelial-like HCC cells through repression of E-cadherin expression, whereas FGF19 knockdown enhances E-cadherin expression and hence diminishes EMT traits in mesenchymal-like HCC cells, suggesting FGF19 exerts its tumor progressing functions as an EMT inducer. Interestingly, depletion of FGF19 cannot abrogate EMT traits in the presence of GSK3β inhibitors. Furthermore, FGF19-induced EMT can be markedly attenuated when FGFR4 is knocked out. These observations clearly indicate that FGFR4/GSK3β/β-catenin axis may play a pivotal role in FGF19-induced EMT in HCC cells. As FGF19 and its specific receptor FGFR4 are frequently amplified in HCC cells, selective targeting this signaling node may lend insights into a potential effective therapeutic approach for blocking metastasis of HCC.

Identification of dysfunctional biological pathways and their synergistic mechanism in hepatocellular carcinoma process

BackgroundHepatocellular carcinoma (HCC) is a lethal and prevalent cancer worldwide. This study was conducted to investigate dysfunctional pathways and their synergistic mechanism in the HCC process. MethodsWe downloaded transcriptome profiling data (GSE25097) from the Gene Expression Omnibus (GEO) database, including 6 healthy liver samples, 40 cirrhosis samples, 243 adjacent non-tumor samples, and 268 HCC samples. Robust Multi-Array (RMA) in R software was employed to preprocess the downloaded dataset, and Student's t-test (FDR less than 0.001) was performed to identify the differentially expressed genes (DEGs) between 4 sample groups. Then, pathway enrichment analysis (FDR less than 0.05) based on iSubpathwayMiner was performed. Furthermore, we performed collaborative analysis on these pathways through calculating the Jaccard index, and crosstalk networks were constructed and visualized by Cytoscape. ResultsTotally, 4617, 9517, and 12,479 DEGs were identified between healthy liver and cirrhosis samples, cirrhosis and adjacent non-tumor samples, and adjacent non-tumor and HCC samples, respectively. Furthermore, a total of 26 crosstalks involving 13 pathways, 78 crosstalks involving 54 pathways, and 86 crosstalks involving 52 pathways were identified through the DEGs between healthy liver and cirrhosis samples, cirrhosis and adjacent non-tumor samples, and adjacent non-tumor and HCC samples, respectively. Moreover, 5 dysfunctional pathways were found to co-exist in the three processes of HCC. Among them, 3 dysfunctional pathways have collaborative relationship, including Staphylococcus aureus infection, leishmaniasis, and Chagas disease. ConclusionsIn this study, dysfunctional pathways in the HCC process and crosstalks between these pathways were investigated for the first time, providing new insight into the potential mechanisms of HCC.

Role of hepatitis B virus integration in evolution of hepatocellular carcinoma

Hepatitis B virus(HBV)-human DNA integration is a commonly seen event in the evolutionary process of hepatocellular carcinoma(HCC).HBV chronic infection induces HBV mutation,HBV integration and the inflammatory microenvironment alternation in the hosts,which provides a evolutionary soil for the hepatocarcinogenesis.The second generation sequencing technique provides a better approach for understanding the HBV integration mechanism in HCC.This review focuses on the features of HBV integration and influences of HBV integration on human,virus genomes,and discusses the association of HBV integration with HCC.

Expression and Role of ICAM-1 in the Occurrence and Development of Hepatocellular Carcinoma

Intercellular adhesion molecule-1 (ICAM-1) is a member of the immunoglobulin superfamily, its main function being to participate in recognition and adhesion between cells. ICAM-1 is considered closely related to occurrence, development, metastasis and invasion process of hepatocellular carcinoma (HCC). A variety of inflammatory cytokines and stimulus affect its expression through the nuclear factor-kappa B (NF-κB) signal transduction pathway. In the initial stage of inflammation, hepatocirrhosis and tumor development, ICAM-1 is expressed differently, and has varied effects on different cells to promote occurrence of malignancy and metastasis. ICAM-1 has diagnostic significance for AFP-negative or suspected HCC, and may be a prognositic significance. It is thus widely used in studies as a biomarker which reflects cancer cells metastasis as well as curative effect of drugs. Many new treatments of HCC may be based on the effects of ICAM-1 on different levels of function.

Structural studies on the CXCR7 decoy receptor

Motivations. Recent studies have revealed that the chemokine CXCR7 plays an important role in cancer development. However, little is known about the effect of CXCR7 on the process of hepatocellular carcinoma (HCC) cell invasion and angiogenesis. In particular, CXCR7 was overexpressed in HCC tissues. The CXCR7-receptor is a transmembrane protein that binds two chemokines, CXCL11 and CXCL12. It has the same basic structure of the other chemokine receptors composed by 7 helices and some Cys residues within the primary sequence to stabilize the tertiary structure through intermolecular disulfide bonds. The signal transduction of chemokines occurs, usually via the G proteins whereas CXCR7 is a decoy and behaves as the receptor Duffy. In particular, it does not bind to G protein, and then does not activate its metabolic pathway. When CXCR7 binds to CXCL12, it can form homodimers or heterodimers from the moment that tends to bind also to CXCR4. In the case of heterodimer (CXCL12/CXCR4/CXCR7), CXCR7 changes the conformation of the CXCR4/G-protein complex and repealing signaling. The activation of CXCR4 by CXCL12 leads to the activation of signaling through PI3K/AKT, IP3, and MAPK pathways, that promote cell survival, proliferation and chemotaxis. In addition, the pathway of the β-arrestin may be activated by GRK that leads to the internalisation of CXCR4. When CXCR7 alloy CXCL12, the mobilization of intracellular Ca2 + classical does not occur, and the activation of β-arrestin may lead to the scavenging.CXCL12. In tumor cells CXCR7 can also activate the signaling via PLC / MAPK that increases the survival of the cells. Therefore, to study the structure-function relationships in CXCR7, we have modeled its three dimensional structure as well as studied the energetic stability of the protein by molecular dynamics simulations. Finally, its complex with CXCL11 and CXCL12 has been simulated by docking methods. Methods. The three-dimensional model of human CXCR7 was performed by a comparative modeling strategy using as template the structure of the human CXCR4, recently published, and that of bovine rhodopsin using MODELLER9v5 program. Models have been evaluated in terms of stereochemical, structural packing and energetic quality. PatchDock web server was used to model the complexes between CXCR7 and the two ligands, CXCL11 and CXCL12, using for CXCR7 our model obtained by comparative modeling, for CXCL11 and CXCL12 their experimental structures. The complexes were analyzed by ''Cocomaps Server'' to identify the amino acids at the interface and to evaluate their solvent accessibility. The presence of putative H-bonds was calculated with Hbplus program. Results. Our studies show that CXCR7 model presents seven trans-membrane helices, a long N-terminal segment and three loops (i.e. loops 1, 2 and 3) in the extracellular region and, moreover, other three loops and a C-terminal loop in the cytoplasmatic region can also be observed. This model had a Prosa Z-score of -2.19 and 86.54% of residues in most favored regions. In particular, the extracellular loop 2 and the cytoplasmatic C-terminal region comprise two short β-strands and a short helix, respectively, in agreement with the experimental structure of CXCR4. A comparison between the secondary structures of CXCR7 and CXCR4 models shows seven well conserved trans-membrane helices and only few changes in helix length. Furthermore, we have modeled two complexes between CXCR7 and two chemokines, CXCL11 and CXCL12. For these complexes, we evaluated the interaction residues, the related contact maps and the number of inter-chain H-bonds. In general, the analysis of the two complexes shows that the interacting regions are located in the N-terminal region as well as in the loops 2 and 3 of CXCR7. We have also analyzed the physico-chemical properties of interaction residues of CXCR7 in the two complexes: i) some positively and negatively charged residues are present in the N-terminal while in the CXCR7/CXCL11 complex we also found also an aromatic residue; ii) the loop 2 presents a negatively charged residue (Glu) and an aromatic residue (Tyr); iii) the loop 3 shows a single aromatic residue (Phe). These observations suggest that the predominant interactions found in the models between CXCR7 and the two chemokines are on hydrophobic and electrostatic basis. In particular, CXCR7 presents the highest affinity for CXCL12 in terms of binding energy, and of number of H-bonds, of charged and hydrophobic interaction residues, and of salt bridges. Moreover, the presence of stacking interactions is also in good agreement with experimental studies which indicated that the affinity of CXCL12 for CXCR7 is approximately ten times higher than the affinity of CXCL12 for CXCR4.

Decision making in elderly HCC

Background The decision making process in hepato-cellular carcinoma (HCC) relies on multiple factors. Hepatic surgery, in fact, has to take into consideration a patient’s general medical condition, performance status, underlying liver disease, functional hepatic reserve. All these factors affect hepatic metabolism, flux and regeneration mechanism conditioning future liver remnant volume. What is more, hepatic regeneration is delayed, even if not completely impaired, in the elderly. Nevertheless age is not always properly taken into account.