Effects Of Pim-3 Antisense Oligodeoxynucleotide On Proliferation And Apoptosis Of Human Hepatocellular Carcinoma Cell Line SMMC-7721

Objective:Pim-3; a member of a proto-oncogene Pim family with serine/threonine kinase activity, was expressed in human hepatocellular carcinoma tissues and cell lines but not in normal hepatocytes, its contribution to hepatocarcinogenesis remains unknown. To investigate effects of Pim-3 fully phosporothioated antisense oligodeoxynucleotide (Pim-3) ASODN on proliferation and apoptosis of human hepatocellular carcinoma SMMC-7721 cell.Methods:Pim-3 ASODN was transfected into SMMC-7721 cells by Lipofect AMINE 2000. Cell proliferation was detected by MTT.The expression of Pim-3 was detected by Reverse Transcription-Polymerase Chain Reaction(RT-PCR) and Western blot . Flow cytometry(FCM)was employed for examining the cell cycle of the transfected SMMC-7721 cells.Results:MTT assay demonstrated that Pim-3 ASODN could significantly inhibit the proliferation of the cells in both time and concentration dependent manner, compared to missense ODN, liposomes and DMEM medium(P<0.05).RT-PCR and Western blot analysis showed the mRNA and protein expression of Pim-3 ASODN cells compared with three control groups was significantly reduced(P<0.05).FCM results showed :Compared with the control group cells, the apoptosis of the SMMC-7721 cell transfected by Pim-3 ASODN significantly increased and cell cycle obviously arrest at G1 period(P<0.05).Conclusion:Pim-3 ASODN is able to inhibit the proliferation of SMMC-7721 cell and induce the cell apoptosis,which might provide a new path for hepatocellular carcinoma gene therapy.