Survivin反义核酸诱导卵巢癌细胞凋亡及逆转多西他赛耐药机制的实验研究  

Experimental study on apoptosis of ovarian cancer cells and reversal docetaxel drug resistance induced by Survivin antisense RNA

作　　者：马荣[1] 陈曦海[2] 唐丽萍[1] 耿晓星[1] 张云艳[1] 张英蕾 王晶[1] 

机构地区：[1]哈尔滨医科大学附属肿瘤医院妇科,黑龙江哈尔滨150040 [2]哈尔滨医科大学附属第四医院普外科,黑龙江哈尔滨150001 [3]黑龙江省人才中心,黑龙江哈尔滨150001

出　　处：《第四军医大学学报》2006年第17期1616-1619,共4页Journal of the Fourth Military Medical University

基　　金：黑龙江省教育厅课题(10551152)

摘　　要：目的构建Survivin反义真核表达载体(anti-pcD-NA3-svv),探讨其对卵巢癌的凋亡诱导作用、对多西他赛的化疗增敏作用及逆转耐药的机制.方法采用Survivin反义核酸瞬时电转染法转染卵巢癌细胞系Skov3,筛选出阳性细胞株.以Westernblot检测Survivin蛋白的表达,用流式细胞仪检测其对Skov3的凋亡诱导作用,采用MTT法检测其对多西他赛的化疗增敏作用,并检测实验组细胞(Skov3-SVVanti)MDR-1mRNA的表达.结果稳定表达anti-pcDNA3-svv的Skov3-SV-Vanti其Survivin蛋白表达比未转染者明显降低,Skov3-SV-Vanti组细胞流式细胞仪检测显示其凋亡率为19%,对多西他赛的IC50值为(13.3±2.25)ng/mL,而对照组(Skov3)为(53.2±2.45)ng/mL,差异具有明显的统计学意义(P<0.01).Skov3-SVVanti组MDR-1mRNA表达较Skov3组明显减少(P<0.01).结论Survivin反义核酸可诱导卵巢癌细胞凋亡,并通过降低MDR-1mRNA表达增强多西他赛化疗敏感性.AIM： To establish Survivin antisense RNA and explore its effect on the apoptosis of ovarian cancer cells and the sensitivity to docetaxel and its mechanism of reversing docetaxel drug resistance. METHODS： Survivin antisense eukaryotic vector （anti-pcDNA3-svv） was transferred into Skov3 cells by electroporation and positive clone was screened out. Survivin protein was detected by Western blot; effect of apoptosis inducement was observed by flow cytometry; sensitivity to docetaxel was examined by MTT; expression of MDR-1 mRNA was detected. RESULTS： Survivin protein of positively cloned cells were reduced obviously as compared with that of non-transfered cells. Terminal apoptotic change was observed by flow cytometry and the apoptosis rate was 19% ; IC50 of experimental group to docetaxel was （13.3 ± 2.25 ） ng/mL and control group was （53.2 ± 2.45 ） ng/mL, so the statistical difference was significant （ P 〈 0. 01 ）. Expression of MDR-1 mRNA of Skov3-SVVanti group was reduced obviously as compared with that of Skov3 group （ P 〈 0. 01 ）. CONCLUSION： Survivin antisense RNA can induce ovarian cancer cell apoptosis and increase the sensitivity to docetaxel by reducing the expression of MDR-1 mRNA.

关 键 词：SURVIVIN反义核酸 卵巢肿瘤 凋亡 多西他赛 多药耐药基因 

分 类 号：R737.31[医药卫生—肿瘤]