[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.


CONCLUSIONS
Transfection of survivin ASODN could induce hepatocellular carcinoma cells apoptosis effectively through activated p38MAPK-caspase-3 signal pathway sequentially.
Chen
Tao
T
Center of General Surgery, General Hospital of Chengdu Command, Chengdu 610083 China.
Tian
Fu-Zhou
FZ
Cai
Zhong-Hong
ZH
Yin
Zhi-Liang
ZL
Zhao
Tie-Jun
TJ
D000951:Antigens, Neoplasm; D017209:Apoptosis / Q000187:drug effects*; D006528:Carcinoma, Hepatocellular / Q000473:pathology*; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D045744:Cell Line, Tumor; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D008113:Liver Neoplasms / Q000473:pathology*; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000494:pharmacology*; D009363:Neoplasm Proteins; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000494:pharmacology*; D015398:Signal Transduction; D000077022:Survivin; D014162:Transfection; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
D000951:Antigens, Neoplasm; C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D016376:Oligonucleotides, Antisense; D000077022:Survivin; D048051:p38 Mitogen-Activated Protein Kinases; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases