| Chronic hepatitis B virus infection is a major risk factor for liver cirrhosis and hepatocellular carcinoma in China.Splice variants of HBV can be generated from HBV pregenomic RNA by splicing and reverse transcription.The 2.2 kb spliced defective HBV genome are the most common and identified as singly or doubly spliced isoforms depending on the splicing pattern.Both of them can be detected in HBV-infected liver tissues and has been shown to correlate with chronicity or persistence of HBV infection,liver fibrosis.Previous studies in our laboratory have confirmed that the 2.2 kb HBV doubly spliced protein,HBDSP,could transactivate some proto-oncogenes and HBV regulatory elements.However,the pathogenesis of HBDSP in liver cancer is still unclear.The present study focus on the invasion and metastasis of HBDSP in hepatocellular carcinoma cells.Primarily,HBDSP was overexpressed by infection of Huh7 or HepG2 cell lines with recombinant adenovirus.Would healing assay,transwell migration and invasion assay,CCK8 assay were subjected to HBDSP-overexpressed cells.Results showed that HBDSP could promote invasion and metastasis of Huh7 or HepG2 cells.Epithelial-mesenchymal transition(EMT)markers including E-cadherin,N-cadherin,Vimentin,Snail were further detected to show that over expression of HBDSP in Huh7 or HepG2 cells could induce EMT.Then,high throughput sequencing were carried out to explore the potential pathogenesis of HBDSP in Huh7 cells by RNA-seq technology.The results showed that 224 genes were up-regulated and 266 genes were down-regulated.11 up-regulated genes and 4 down-regulated genes showed a significant difference.Moreover,gene functional annotation analysis showed that these genes mainly enriched in adherens junction,cell-substrate adherens junction,anchoring junction,focal adhesion,which can be related with tumor invasion and metastasis.These results may establish foundation to reveal the mechanism of HBDSP leading to the occurrence and development of HCC. |
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