Survivin反义寡核苷酸诱导人卵巢癌耐药细胞株COC_1/DDP凋亡的实验研究

Apoptosis of drug-resistant human ovarian cancer cell line COC_1/DDP induced by survivin antisense oligonucleotides.

目的 :探讨经脂质体介导的survivin反义寡核苷酸诱导人卵巢癌耐药细胞株COC1/DDP凋亡的作用。方法 :将脂质体介导的survivin反义寡核苷酸 (survivin ASONs)作用于COC1/DDP细胞 ,采用一步法RT PCR检测survivin、bcl 2mRNA的表达 ,并进行细胞核DNA梯形电泳 ,检测caspase 3活性 ,流式细胞仪检测细胞凋亡率及细胞周期。结果 :经survivin ASONs作用后COC1/DDP细胞中survivin等抗凋亡基因的表达下降 ,细胞核DNA电泳呈现凋亡细胞特有的梯状条带 ,caspase 3活性增加且呈时间依赖性 ,细胞凋亡率达33.18% ,细胞周期G2 /M及S期明显下降而多阻滞于G0 /G1期 ,与对照组差异有显著性(P <0 .0 5 )。结论 :survivin ASONs反义寡核苷酸能诱导人卵巢癌耐药细胞株COC1/DDP细胞凋亡 。

Objective:To explore the apoptosis of drug-resistant human ovarian cancer cell line COC1/DDP induced by liposome-survivin antisense oligonucleotides.Methods:The expressions of apoptosis associated genes(survivin and bcl-2)were studied by reverse transcription-polymerase chain reaction(RT-PCR).The caspase-3 activities were measured by western blot.The apoptosis rate,cell cycle profiles were detected by flow cytometry(FCM).Results:The mRNA expressions of anti-apoptosis genes(survivin and bcl-2) were down-regulated.DNA electrophoresis showed DNA ladder which was the characteristic change of apoptosis.The caspase-3 activity was significantly higher than that of control group,which showed time-dependent(P<0.05).Apoptosis rate reached 33.18% much higher than that of untreated group.Cell cycle profiles changed a lot,G2/M and S phase decreased while G0/G1 increased.Conclusion:Survivin antisense oligonucleotides could effectively induce apoptosis of drug-resistant human ovarian cancer cell line COC1/DDP.