Apoptosis in hepatoma cells induced by antisense oligodeoxynucleotide against survivin / 中华肝脏病杂志

ABSTRACT

<p><b>OBJECTIVES</b>To investigate the apoptosis induced by antisense oligodeoxynucleotide (ASODN) against survivin and the mechanisms after the hepatocellular carcinoma SMMC-7721 cells transfected with the ASODN.</p><p><b>METHODS</b>The ASODN was transfected into SMMC-7721 cells mediated by liposomal reagent. The changes of cell cycle and apoptotic rate were detected by flow cytometry. The changes of cell skeleton was observed through confocal microscope. The activity of p38MAPK and caspase-3 were detected by immuno-precipitation and kinase activity assess methods, respectively.</p><p><b>RESULTS</b>There were control, sense control, 400, 600, 800, and 1 000 ng/ml ASODN groups (I - VI). The apoptotic rats were 0.70%, 0.76%, 2.43%, 7.82%, 23.11%, and 31.35% in groups I - VI, respectively, which in the ASODN-transfected groups were higher than that in the control group (t<or=-9.6, P<0.01). The cell cycle in the ASODN-transfected groups stopped at G2/M phase. The well-arranged structure of microfilament was broken or destroyed, and the average fluorescent intensity of actin were 189.69+/-6.68, 184.23+/-8.76, 173.14+/-8.15, 99.48+/-6.57, 76.69+/-10.05 and 63.80+/-6.79 in groups I - VI, respectively), which in groups IV - VI decreased markedly, compared with that in the control group (t>or=20.9, P<0.01). The activity of p38MAPK increased significantly, when the ASODN was transfected at the concentration of 600 ng/ml or more, so did the caspase-3 activity (the p38MAPK and caspase-3 activity in groups I - VI were 7.03, 7.07, 13.47, 16.37, 43.97, 47.87 and 0.015+/-0.010, 0.014+/-0.002, 0.026+/-0.003, 0.042+/-0.001, 0.093+/-0.001, 0.100+/-0.001, respectively).</p><p><b>CONCLUSIONS</b>ASODN targeting at survivin mRNA can induce G2/M stop, activate p38MAPK and caspase-3. The activated caspase-3 destroys the cell skeleton microfilament system, resulting in apoptosis.</p>