| Objective:TM6SF2 plays an important role in chronic liver disease and lipid metabolism,but its role in hepatocellular carcinoma is remain unknown.In this study,TCGA,the current mainstream tumor database,was firstly used to explore the expression of TM6SF2 in hepatocellular carcinoma and its clinical significance.Meanwhile,the biological function of TM6SF2 in hepatocellular carcinoma and the mechanism of ceRNA regulatory network composed of LncRNA and miRNA were analyzed by combining bioinformatics technology.Finally,huh-7 cell line was stably transfected with shRNA targeting TM6SF2 to reduce the expression of TM6SF2 in huh-7 and explore its effect on huh-7 cell proliferation.Methods:Online tools were used to dig up TCGA tumor database data,analyze the expression of TM6SF2 in hepatocellular carcinoma tissues,and explore the influence of TM6SF2 on the survival and prognosis of liver cancer patients.The online tool Broad GDAC Firehose was used to download RNA-seq data and clinical information of patients with HCC from TCGA,and analyze the relationship among TM6SF2 and patients' general clinical characteristics,pathological progress and primary hepatocyte pathogenic risk factors.Meanwhile,the coding genes associated with TM6SF2 expression in hepatocellular carcinoma were analyzed,and the GO enrichment analysis,KEGG pathway pathway analysis and PPI protein interaction network analysis were conducted by bioinformatics methods to predict the potential biological functions of TM6SF2.TCGA tumor database data were mined using online tools to analyze miRNA and lncRNA associated with TM6SF2 expression in hepatocellular carcinoma,respectively.Bioinformatics method was used to explore the potential regulatory mechanism between TM6SF2,LncRNA and LncRNA in hepatocellular carcinoma based on the analysis of "TM6SF2-mirna-lncrna" ceRNA.Finally,lentivirus was used to stably transfect shRNA targeted at TM6SF2 in huh-7 cells to construct a cell model with stable and low expression of TM6SF2.RT-PCR method was used to test the expression level of TM6SF2 mRNA,so as to detect the knockdown efficiency of TM6SF2 gene in the cell model.Huh-7 cells with stable and low expression TM6SF2 and control huh-7 cells were tested by cck-8 cell proliferation detection kit.SPSS 21.0 version and GraphPad Prism 7 version were used for all statistical methods and mapping.Results:The expression of TM6SF2 in hepatocellular carcinoma was significantly reduced compared with that in normal liver tissues(P<0.01).In patients with hepatocellular carcinoma,the median survival time of the group with low TM6SF2 expression was reduced(HR=1.432,log-rank p=0.0471).TM6SF2 was associated with BMI,pathological staging,histological grade and risk factors of primary hepatocellular carcinoma(P<0.05).There are a total of 49 coding genes expressed in the hepatocellular carcinoma tissues related to TM6SF2.GO enrichment analysis found that the functions of the genes related to TM6SF2 expression were mainly concentrated in the aspects of fatty acid synthesis,fatty acid ligase activation,thrombin regulation,etc.(P <0.05).Analysis of KEGG pathway pathway revealed that the expression related genes were mainly concentrated in alanine metabolism,PPAR signaling pathway,bile secretion and other signaling pathways(P <0.05).Study on the HCC signaling pathway have found that the expression related genes of TM6SF2 are involved in the hepatocyte growth cycle,liver fibrosis and early HCC.PPI protein interaction network analysis suggests that the APOC3 and HAPLN4 TM6SF2 direct effect(P < 0.05).Candidate miRNA that regulate TM6SF2 in hepatocellular carcinoma were screened,including miRNA-149-3p,miRNA-134-3p,miRNA-3689b-3p(P<0.05,FDR<0.05).Meanwhile,LncRNA related to TM6SF2 expression in hepatocarcinoma were analyzed(P<0.01),and the candidate LncRNA that interact with candidate mirnas were predicted,namely MIR194-2HG,GABPB1-AS1,LIC00261 and MAGI-AS3(P< 0.05).Candidate miRNA and LncRNA participate in the construction of TM6SF2 ceRNA regulatory network.TM6SF2-targeted shRNA was transfected into Huh-7 cells successfully and the TM6SF2 stable knockdown Huh-7 cells were selected.The knockdown efficiency of TM6SF2 gene was about 80%.The results of cck-8 method showed that the low expression of TM6SF2 inhibited the proliferation of huh-7 cells(P<0.05).Conclusion:TCGA database and bioinformatics analysis showed that TM6SF2 was low expression in hepatocellular carcinoma tissues,and the median survival time of patients with low expression of TM6SF2 was reduced.Moreover,TM6SF2 was associated with BMI,pathological stage,grade and risk factors of primary liver cancer,and was a potential tumor suppressor.However,in cell experiments,Knock-dowm TM6SF2 inhibited the proliferation of huh-7 cells.The biological functions of TM6SF2 mainly focus on substance metabolism,suggesting that substance metabolism plays an important role in the pathological process of tumor.We explored LncRNA and miRNA that have potential effects on TM6SF2 and provided theoretical support of tumor data and bioinformatics for the role and mechanism of TM6SF2 in the pathological development of hepatocellular carcinoma. |
PDF Full Text Request