反义Survivin核酸诱导凋亡及逆转胃癌耐药机制的实验研究

Experimental study on inducing apoptosis and reversing drug resistance of gastric cancer cells SGC7901 by antisense Survivin RNA

目的:探讨反义Survivin核酸(anti-pcDNA3-svv)对胃癌细胞系SGC7901的凋亡诱导、对泰索帝的化疗增敏作用及对化疗耐药的逆转作用.方法:采用脂质体转染法转染胃癌细胞系SGC7901,并筛选阳性克隆,以Westernblot检测Survivin蛋白的表达,RT-PCR检测MDR-1mRNA的表达,用透射电镜及TUNEL法观察并检测转染后SGC7901的凋亡变化,采用MTT法检测转染细胞对泰索帝的敏感性.结果:转染anti-pcDNA3-svv的SGC7901细胞系(SGC7901-SVVanti)其Survivin蛋白比未转染者明显降低,仅为其29.9%(P<0.01);透射电镜下可见转染组细胞呈凋亡晚期变化,TUNEL显示转染组与未转染组AI分别为0.241和0.083(P<0.01);转染组MDR-1mRNA较未转染组明显降低,转染组与未转染组MDR指数分别为:0.196±0.013和3.126±0.019(P<0.01);转染组对泰索帝的IC50值为16.7±1.98μg/L,而未转染组为55.7±1.89μg/L,差异具有显著的统计学意义(P<0.01).结论:反义Survivin核酸可诱导SGC7901细胞凋亡,增强泰索帝化疗敏感性,逆转耐药.

AIM： To explore the effect of antisense Survivin RNA on the apoptosis, chemosensitivity to taxotere and drug resistance of gastric cancer cells SGC7901. METHODS： SGC7901 cells were transfected with antisense Survivin eukaryotic vector （antipcDNA3-svv） by lipofectamine, and the positive clones were screened out. Survivin protein and multi-drug resistance-1 （MDR-1） mRNA were determined by Western blot and reverse transcription polymerase chain reaction （RT-PCR）.The apoptosis of SGC7901 cells was observed by electronic microscopy and TUNEL method, and the sensitivity of the cells to taxotere was examined by MTr assay. RESULTS： The expression of Survivin protein in the transfected SGC7901 cells （29% of latter） was decreased obviously in comparison with that in the non-transfected cells （P 〈 0.01）. SGC7901-SVVanti cells （transfected） presented terminal apoptosis, and TUNEL showed that the apoptosis index （AI） was 0.241, which was higher as compared with that of non-transferred cells （0.083） （P〈0.01）. MDR-1 index was 0.196±0.013 and 3.126±0.019 in the transfected and nontransfected cells, respectively （P 〈 0.01）. The IC50 of taxotere was 16.7±1.98 and 55.7±1.89μg/L in the transfected cells and non-transfected ones, respectively （P〈0.01）. CONCLUSION： Antisense Surivivin RNA can induce apoptosis, increase the chemosensitivity to taxotere and reverse the drug resistance of SGC7901 cells.