Antisense oligonucleotide targeting survivin transfecting the hepatocellular cell could induce the activity of caspase-3

Objective:To investigate the path of apoptosis induced by antisense oligonucleotide(AS ODN) targeting survivin on hepatocellular cell.Methods: Positive expression of survivin had been confirmed in SMMC-7721 cell line. Antisense oligonucleotide targeting survivin was designed and composed. Twenty-four hours before transfection, SMMC-7721 cell were cultured in 6-well culture dish.Cultured cells were divided into 6 groups:①control group;②lipofectin group;③ODN group;④200nmol/L AS ODN group;⑤400nmol/L AS ODN group; ⑥600nmol/L AS ODN group. After transfection for 20 hours, cultured cells were harvested to continue next tests.①Cell form was observed with invert microscope;②Survivin protein expression was detected by western blot;③Apoptotic index(AI) and proliferative index(PI)were examined by flow cytometry;④Activity of caspase-3 was detected by western blot. Results: ①Expression of survivin in AS ODN groups were significantly decreased than that of the control, lipofectin and ODN group;②Abnormal form of cells was observed in AS ODN transfected groups;③The PI of AS ODN groups were significantly higher than that of other groups;④The PI of AS ODN groups were significantly lower than that of other groups;⑤Activity of caspase-3 was increased in AS ODN groups but not in other groups. Conclusion:Antisense oligonucleotide targeting survivin could decrease the expression of survivin, induce hepatomacellular apoptosis and inhibit cell proliferation.