Promotion effects of survivin-ASODN on apoptosis of SMMC-7221 cell and its mechanism / 吉林大学学报(医学版)

ABSTRACT

Objective To study the influence of survivin targetedly inhibited with antisense oligonucleotide (ASODN)technique on the apoptosis of hepatoma carcinoma cells SMMC-7221,and to clarify the mechanism of promotion effect of survivin-ASODN on the apoptosis of SMMC-7721 cells.Methods The ASODN sequence of survivin marked by FAM fluorescein was designed and synthized. The SMMC-7721 cells were transfected by different concentrations (100,200,300,400,and 600 nmol· L-1 )of survivin-ASODN (ASODN transfection groups),at the same time blank control group and blank liposome control group and sense oligonucleotide (SODN) control group were set up.The apoptotic rates and the changes of cell cycle of the SMMC-7721 cells 24,48,and 72 h after transfected with different concentrations of survivin-ASODN were detected by FCM. The expression levels of survivin were measured by Western blotting method.Results Compared with each control group,24 h after transfection,the apoptotic rates of survivin-ASODN transfected SMMC-7221 cells were increased,the growth of cells was inhibited (P<0.05),and the effects had time-dose dependent tendency.48 h after transfection,the hypodiploid apoptotic peak appeared in ASODN transfection groups before G1 phase, the number of the cells at G0/G1 phase was decreased (P<0.05)and the number of the cells at G2/M phase wsa increased (P<0.05). Compared with each control group,the survivin expression levels in the SMMC-7721 cells in ASODN transfection groups were decreased (P<0.05 ), and the effects of survivin-ASODN was time-dose dependent (P<0.05 ). Conclusion Survivin-ASODN can block the expression of survivin in SMMC-7721 cells and inhibit the proliferation of SMMC-7721 cells by changing the cell cycle and increasing apoptosis in a time-dose dependent manner.