Preliminary Study On The Ability Of HBx Transfection To Promote Invasion And Metastasis Of Hepatocellular Carcinoma Cells And The Difference Of LncRNA Expression Profile

Objective: To study the effect of HBx on the invasion and metastasis of human hepatocellular carcinoma cell line HepG2 and Huh7 by transient transfection of HBx gene and the differential expression profile of lncRNA before and after transfection.Therefore,it will provide a new idea for the diagnosis and treatment of hepatocellular carcinoma with HBV infection.Methods: Two hepatocellular carcinoma cell lines,HepG2 and Huh7,were cultured in vitro,and divided into three groups: blank group,pcDNA3.1 transfection group and pcDNA3.1-HBx transfection group.RT-PCR was used to detect the expression of HBx mRNA in two hepatocellular carcinoma cell lines.Transwell assay was used to detect the effect of HBx on the invasiveness of hepatocellular carcinoma cells.The effect of HBx on the metastatic ability of hepatocellular carcinoma cells was detected by scratch test.The differential expression profile of lncRNA in HepG2 cells was detected by Illumina HiSeq?2000 sequencer and DEseq2 software before and after HBx transfection.QRT-PCR was used to verify the expression of lncRNA mRNA in HepG2 cells before and after HBx transfection.Results: 1.The results of transwell assay showed that the number of invasive cells in HepG2 and Huh7 cells was significantly increased in pcDNA3.1-HBx transfection group.The results of scratch test showed that the migration distance in HepG2 and Huh7 cells was significantly longer in pcDNA3.1-HBx transfection group.2.The results of RNA sequencing showed that there were 29 differentially expressed mRNAs in hepatocellular carcinoma cells before and after HBx transfection,of which 17 were up-regulated and 12 were down-regulated;there were 10 differentially expressed lncRNAs,of which 3 were up-regulated and 7 were down-regulated.3.QRT-PCR results showed that the expression of lncRNA ENST00000582866.1 in pcDNA3.1-HBx group was significantly down-regulated compared with that in pcDNA3.1 group.Conclusion : 1.Transfection of HBx gene can enhance the invasion and metastasis ability of hepatocellular carcinoma cells.2.Differential analysis of lncRNA expression profile in hepatocellular carcinoma cells with transient HBx showed that the differential expression genes of mRNA and lncRNA were 29 and 10 respectively,among which 17 and 3 were up-regulated,12 and 7 were down-regulated respectively.