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[The signal transduction pathway related to hepatocellular carcinoma apoptosis induced by survivin antisense oligonucleotide].

OBJECTIVE
The expression of survivin gene in hepatocellular carcinoma cell line SMMC-7721 was blocked by means of antisense oligonucleotide (ASODN) transfection. To investigate the signal transduction pathway of apoptosis induced by survivin ASODN.


METHODS
survivin ASODN was transfected into SMMC-7721 cells mediated by DOTAP liposomal reagent. The expression of survivin protein and mRNA was detected by western-blot and in situ hybridization method, respectively. Flow cytometer and TUNEL method were used to detect apoptosis. The changes in the expression and activity of p38MAPK and caspase-3 were assessed by western-blot, immuno-precipitation, RT-PCR, and kinase activity assess to study the relationship between the changes of p38MAPK and caspase-3 activity.


RESULTS
The expression of survivin protein and mRNA decreased 84.6% and 69.7% respectively while apoptosis increased from 0.70% to 31.15% after transfected with survivin ASODN. P38MAPK activity increased significantly followed by increasing of caspase-3 activity after survivin ASODN transfection. Both p38MAPK and caspase-3 activity were inhibited after treated with inhibitor of p38MAPK, SB202190.


CONCLUSION
Transfection of survivin ASODN could induce hepatocellular carcinoma cells apoptosis effectively through activated p38MAPK-caspase-3 signal pathway sequentially.