The Expression Distribution And Function Of HCV Non-structural Protein5A Trans-activated Protein9

Background And Objective:NS5ATP9is a trans-activator gene of HCV NS5A protein, screened by suppression subtractive hybridization (SSH). The present study shows that NS5ATP9gene is associated with a variety of tumor tissue. It involved in DNA repair, cell signaling pathways and the regulation of cell proliferation. But the reports about NS5ATP9expression in human tissues and cell lines and its subcellular location are inconsistent. In this study, we conducted NS5ATP9expression, distribution, and explore preliminaryly NS5ATP9gene function in different cell lines, mouse tissues and organs, intracellular localization and adjacent tissues of the liver and cancer aspects.Methods:(1) The expression of NS5ATP9in different liver cancer cell lines in mRNA levels and protein levels were examined by Real-time quantitative PCR and Western Blot.(2) The mRNA expression of NS5ATP9in human hepatocellular carcinoma and adjacent noncancerous tissues were deetected by real-time PCR.(3) The mitochondrial marker protein recombinant vector pCMV-PDHAl-N-RFP and the recombinant plasmids pEGFP-NS5ATP9were co-transfected into HepG2cells, and the subcellular localization of NS5ATP9were observed by fluorescence microscope.(4) The pEGFP-Nl-NS5ATP9recombinant plasmids were transfected into HepG2cells and the biological function of the NS5ATP9were explored by microarray expression profiling.Results:(1)The expression of NS5ATP9in Huh7, QYH7701, L02, SMMC7721and HepG2liver cancer cell lines in mRNA levels were2.46,5.74,6.06,12.04and19.29times compared to Be17402liver cancer cell lines.(2) The mRNA expression of NS5ATP9in human hepatocellular carcinoma were0.26,0.21,0.12,0.47,0.84times lower than adjacent noncancerous tissues.(3) The mitochondrial marker protein recombinant vector pCMV-PDHAl-N-RFP and the recombinant plasmids pEGFP-NS5ATP9were co-transfected into HepG2cells, and the subcellular localization of NS5ATP9were mainly distributed in the liver the cell nucleus.(4) NS5ATP9protein in different cell lines detectioned by Western blot showed that NS5ATP9mainly existed in the nucleus.(5) Biological function of the NS5ATP9explored by microarray expression profiling shows that the most target genes connected with immune response and signal transduction, most of them were located in cell membranes or associated with cell membrane, most genes of knownen functional were connected with ion channels activity and enzyme inhibitors activity.Conclusions:(1)The expression of NS5ATP9in different liver cancer cell lines in mRNA levels were low,but hightest in HepG2cell lines compared with others (2) The mRNA expression of NS5ATP9in human hepatocellular carcinoma were lower than adjacent noncancerous tissues,NS5ATP9maybe participated in the progress of HCV carcinogenic mechanism as a Tumor-suppressor genes (3) The subcellular localization of NS5ATP9were observed by fluorescence microscope showed that its position was in the nucleus.(4) Biological function of the NS5ATP9explored by microarray expression profiling shows that the function may be through the adjustment of membrane protein on ion channel activity and signal transduction mechanism to achieve its to immune response of the adjustment, this may be related with tumor mechanism cause by HCV...