Ma Rong, Chen Xi-Hai, Zhang Qi-Fan, Zhu Lei, Tang Li-Ping, Wang Jing, Cheng Hai-Yan, Lei Li-Na
Department of Gynecology, Haerbin Medical University Affiliated Tumor Hospital, Haerbin 150040, China.
Zhonghua Zhong Liu Za Zhi. 2007 Feb;29(2):89-92.
To explore the effects of survivin antisense RNA on apoptosis and chemosensitivity to docetaxel in gastric cancer line SGC7901 cells, and its relation to mdr-1.
survivin antisense eukaryotic vector anti-pcDNA3-svv was transfected into SGC7901 cells by electorporation and positive clone was screened out. survivin protein and mdr-1 mRNA were determined by Western blot and RT-PCR. Apoptosis-inducing effect was examined by electron microscopy. Sensitivity to docetaxel was examined by MTT. Expression of mdr-1 and survivin mRNA were detected in the SGC7901 cells after drug-resisitance induction.
The expression of survivin protein of SGC7901 cells after transfection reduced significantly than that of non-transfected cells. MDR indexes of transfection group and non-transfection group were 0.196 +/- 0.013 and 3.126 +/- 0.019, respectively. The IC50 of transfection group to docetaxel was (16.7 +/- 1.98) microg/L and non-transfection group was (55.7 +/- 1. 89) microg/L, with a statistically significant difference. Expression of survivin mRNA in drug-resistant cells decreased along with the decreasing of mdr-1.
Antisense surivivin RNA can induce apoptosis in gastric cancer cells and increase sensitivity to docetaxel. The reversing mechanism of drug resistance is related with decreasing of mdr-1.
探讨生存素反义RNA对胃癌SGC7901细胞凋亡及多西他赛化疗敏感性的影响及其与mdr-1的关系。
采用电穿孔法将生存素反义真核表达载体anti-pcDNA3-svv转染至SGC7901细胞，筛选阳性克隆。采用蛋白质免疫印迹法和逆转录-聚合酶链反应检测生存素蛋白和mdr-1 mRNA。通过电镜观察细胞凋亡诱导作用。采用MTT法检测对多西他赛的敏感性。检测诱导耐药后的SGC7901细胞中mdr-1和生存素mRNA的表达。
转染后SGC7901细胞生存素蛋白表达较未转染细胞明显降低。转染组和未转染组的多药耐药指数分别为0.196±0.013和3.126±0.019。转染组对多西他赛的IC50为(16.7±1.98)μg/L，未转染组为(55.7±1.89)μg/L，差异有统计学意义。耐药细胞中生存素mRNA表达随mdr-1降低而降低。
反义生存素RNA可诱导胃癌细胞凋亡并增加对多西他赛的敏感性。耐药逆转机制与mdr-1降低有关。
