[Inhibitory effects of survivin antisense oligonucleotide on drug-resistant human ovarian cancer cell COC1/DDP].
OBJECTIVE
Recent studies have shown that survivin is an anti-apoptosis gene, which plays an important role in the carcinogenesis and drug resistance of ovarian cancer. This study was designed to explore the effects of liposome-survivin antisense oligonucleotide (Lip-ASODN) on the growth,apoptosis,and cell cycle distribution of drug-resistant human ovarian cancer cell line COC1/DDP.


METHODS
Survivin-ASODN were transfected into COC1/DDP cells mediated by lipofectin. The proliferation of COC1/DDP cells was assessed by cyto-dynamics and MTT assay. The mRNA expression of survivin was determined by reverse transcription- polymerase chain reaction (RT-PCR). The caspase-3 protein activity was measured by Western blot analysis. The apoptotic rate and cell cycle distribution were estimated by flow cytometry (FCM).


RESULTS
Compared with Lip-SODN and Lip alone groups, the proliferation of COC1/DDP cells after cultured with Lip-ASODN has been significantly inhibited, its inhibitory rate was (68.3+/-6.2)% after cultured for 72 hours (P< 0.05). The mRNA expression of survivin in Lip-ASODN group was significantly decreased, while the caspase-3 activity increased in a time-dependent manner as compared with Lip-SODN and Lip alone groups (P< 0.05). Cell cycle distribution significantly changed in Lip-ASODN group, many cells have been blocked in G0/G1 phase (79.21%), while G2/M phase (4.92%) and S phase (15.87%) decreased. The apoptotic rate of Lip-ASODN group reached 33.18%, which was much higher than those of Lip-SODN and Lip alone groups (P< 0.05).


CONCLUSIONS
Survivin ASODN can inhibit COC(1)/DDP cell proliferation, reduce the mRNA expression of survivin, and induce cell apoptosis.
Ruan
Fei
F
Women's Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, 310006, PR China. rf100@163.com
Xie
Xian-Kuan
XK
Liu
Shao-Yang
SY
D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002945:Cisplatin / Q000494:pharmacology; D019008:Drug Resistance, Neoplasm; D005260:Female; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D008081:Liposomes; D008869:Microtubule-Associated Proteins / Q000096:biosynthesis* / Q000235:genetics; D009363:Neoplasm Proteins; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000494:pharmacology*; D010051:Ovarian Neoplasms / Q000378:metabolism / Q000473:pathology*; D000077022:Survivin; D014162:Transfection
D000970:Antineoplastic Agents; C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D008081:Liposomes; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D016376:Oligonucleotides, Antisense; D000077022:Survivin; D002945:Cisplatin