靶向survivin的siRNA联合顺铂抑制肺腺癌A549细胞增殖的实验研究  

Empirical study on the anti-proliferation effect of siRNA against survivin combined with DDP on human lung cancer cell line A549

作　　者：姚草原[1] 江涛[1] 韩晓黎[1] 李凡[2] 

机构地区：[1]重庆医科大学附属第一医院呼吸内科,重庆400016 [2]重庆医科大学附属第一医院普外科,重庆400016

出　　处：《重庆医科大学学报》2009年第2期178-181,共4页Journal of Chongqing Medical University

摘　　要：目的:研究靶向survivin的siRNA和顺铂联用对肺癌细胞A549的增殖抑制作用。方法:将A549细胞分为空白对照组,阴性对照质粒pSilencer2.0-NC转染组,顺铂处理组,pSilencer2.0-SVV2转染组,pSilencer2.0-SVV2转染+顺铂处理组。转染采用脂质体法。MTT法检测靶向survivin的siRNA和顺铂对A549细胞增殖的抑制作用;RT-PCR法检测A549细胞survivin mRNA转录水平变化情况;流式细胞术和TUNEL法检测A549细胞凋亡情况。结果:MTT法示:pSilencer2.0-SVV2转染+顺铂处理组增殖抑制率为51.38%±1.35%,与其它各组相比抑制率明显增高,具有统计学意义(P<0.05);RT-PCR检测显示空白对照组,阴性对照质粒pSilencer2.0-NC转染组,顺铂处理组survivin mRNA表达无明显变化(P>0.05),pSilencer2.0-SVV2转染组、pSilencer2.0-SVV2转染+顺铂处理组survivin mRNA表达明显下降(P<0.05);流式细胞术和TUNEL法检测pSilencer2.0-SVV2转染+顺铂处理组凋亡率分别为42.48%±2.28%和40.65%±3.53%,与其它各组相比凋亡率明显增高,具有统计学意义(P<0.05)。结论:将靶向survivin的siRNA和顺铂联合应用可以协同抑制A549细胞增殖,共同发挥诱导细胞凋亡作用。Objective： To investigate the anti-proliferation effect of siRNA against survivin combined with DDP on human lung cancer cell line A549.Methods： A549 cells were divided into five different treatment groups： blank control group, pSilencer2.0-NC group, DDP group,pSilencer2.0-SVV2 group,and pSilencer2.0-SVV2＋DDP group. LipofectamineTM 2000 was used to transfect A549 cell. The effects on cell proliferation and apoptosis were analyzed by MTT assay, flow cytometry, and TUNEL respectively .The expression of survivin mRNA was evaluated by RT-PCR experiment. Results： The MTT assay indicated that the anti-proliferation rate of pSilencer2. 0-SVV2＋DDP group was 51.38% ± 1.35% .The anti-proliferation rate of this group was higher than any other group （P〈0.05）.The experiment of RT-PCR showed that the expression of survivin mRNA in pSilencer2.0-SVV2 group and pSilencer2.0-SVV2＋DDP group were lower than that in other groups（P〈0.05 ）, but its level in blank control group, pSilencer2.0-NC group, DDP group had no marked change （P〉0.05）.Flow cytometry and TUNEL indicated that the apoptosis rates of pSilencer2.0-SVV2＋DDP were 42.48% ± 2.28% and 40.65% ± 3.53% respectively, higher than in any other groups（P〈0.05 ）. Conclusion： siRNA against survivin combined with DDP can enhance the inhibitory effect on lung cancer cell line A549 and induce apoptosis in common.

关 键 词：RNA干扰 SURVIVIN基因 肺肿瘤 顺铂 

分 类 号：R734.2[医药卫生—肿瘤]