Analysis And Significance Of MRNA-lncRNA Expression Profiles Of HBx Transfected Different Hepatocellular Carcinoma Cells

Objective:1.To investigate the effect of mRNA and lncRNA expression profiles of HBx transfected hepatocarcinoma cell lines.2.Screening of hepatocarcinoma cell lines HepG2 and Huh7 for lncRNAs with opposite regulation of HBx.3.To investigate the effect of HBx-related ENST00000582866 on the invasion and metastasis of HepG2 and to find potential targets.Methods:1.Using gene overexpression transfection technology to transfer HBx into hepatocellular carcinoma cells to construct a HBV infected cell model.2.The expression levels of mRNA and lncRNA before and after transfection were analyzed based on RNA-Seq,and KEGG and GO enrichment analysis were performed on differentially expressed genes.3.The lncRNA expression profiles in HepG2 and Huh7 cells were overlapped before and after transfection,and the lncRNAs with opposite regulation of HBx in the two groups of hepatocellular carcinoma cells were screened.4.Using gene overexpression and small RNA interference techniques to overexpress or reduce the expression level of ENST00000582866 in hepatocellular carcinoma cells.5.The effects of ENST00000582866 on invasion and metastasis of hepatocellular carcinoma cells were observed by transwell invasion and scratching experiments.6.Prediction of potential targets of ENST00000582866 based on bioinformatics and target verification using q-PCR technology.Results:1.RNA-Seq detected that HBx transfection caused dysregulation of 554 mRNA and 10 lncRNA in HepG2,and 508 mRNA and 62 lncRNA expression in Huh7.2.Coincidence analysis showed that the expression of 8 lncRNAs was reversed after HBx transfection of different lines of hepatocellular carcinoma cells.3.The results of Transwell invasion assay showed that the number of cells in the over-expression group of ENST00000582866 was significantly decreased,while the number of cells in the siRNA-ENST00000582866 group was significantly increased.The scratch test showed that the migration distance in the ENST00000582866 overexpression group was shortened,while the migration distance in the siRNA-ENST00000582866 group was significantly increased.4.The expression level of PLK1 in the ENST00000582866 overexpression group was significantly increased,while the expression level in the siRNA-ENST00000582866 group was significantly decreased.Conclusion:This study was the first to compare the expression profiles of mRNA and lncRNA in HBx transfected HepG2 and Huh7 cells,and preliminarily confirmed that HBx-related lncRNA-ENST00000582866 can inhibit the invasion and migration of hepatocellular carcinoma cell.The possible reason is that ENST00000582866 made the expression of PLK1 is up-regulated to inhibit invasion and migration of hepatocellular carcinoma cell.The above provides effective information for molecular precision treatment of hepatocellular carcinoma cell.