Abstract:

Objective   To construct a short hairpin RNA (shRNA) eukaryotic expression vector targeting to the Livin gene, and explore its influence on chemotherapeutic sensitivity of HepG2 cells. Methods   The Livin shRNA eukaryotic expression vectors pSD11-U6/Neo/GFP/livin were designed, constructed and transfected stably into HepG2 cells. The Livin mRNA and protein relative expression were detected by fluorescence quantitative PCR and Western blotting. MTT was used to evaluate the inhibition rate of HepG2 cells after exposure to cisplatin (2.0mg/L).  ResultsThe DNA sequencing confirmed that the Livin shRNA eukaryotic expression vectors were successfully constructed. Transfection of the vectors could significantly decrease the Livin mRNA and protein relative expression of HepG2 cells (P<0.05). The inhibition rate of cisplatin in transfected cells were obviously higher than those in the untransfected cells (P<0.05). Conclusion    The shRNA eukaryotic expression vectors targeting to the Livin gene are constructed successfully. Transfection of the vectors can effectively decrease the expression of Livin gene in HepG2 cells, and increase the sensitivity of HepG2 cells to chemotherapy.

Key words: Livin gene; RNA interference; Hehapocellular neoplasm