Construction of siRNA eukaryotic expression vectors of PEG10 gene and their apoptosisª²inducing effect on transfected HepG2 cells

    HUANG Jin, LIN Juª²Sheng, DONG Xuª²Yang, CHANG Ying, SONG Yuª²Hu

    Institute of Liver Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China

    ¡¾Abstract¡¿ AIM: To construct the siRNA eukaryotic expression vectors of PEG10 gene and to investigate their apoptosisª²inducing effect on transfected HepG2 cells in vitro. METHODS: Three specific siRNAs of PEG10 were designed, and cloned into the psiRNAª²hH1neo, which were transiently transfected into HepG2 via Lipofectamine 2000. Realtime quantitative PCR was used to measure the PEG10 mRNA expression in HepG2. The cell growth was assessed by MTT assay and the apoptosis was evaluated by flow cytometry and confocal microscopy. RESULTS: Three specific siRNA eukaryotic expression vectors (siRNA1, siRNA2, siRNA3) targeting PEG10 were successfully produced, which could inhibit the expression of PEG10 mRNA to varying degrees. The expression of PEG10 mRNA was remarkably inhibited at 48 h after transfection, and siRNA2 resulted in the highest inhibition rate (93.99%). HepG2 growth was also inhibited significantly (P<0.05), and cell apoptosis rate reached the highest after siRNA2 transfection (66.91%). CONCLUSION: SiRNA eukaryotic expression vectors targeting PEG10 gene can induce HepG2 cell growth inhibition and apoptosis through downª²regulating PEG10. ......