摘要
目的:研究RNA干涉Survivin基因表达对人宫颈癌Hela细胞凋亡及抗肿瘤药物敏感性的影响.方法:将Sur-vivin SiRNA片段通过脂质体的介导转染宫颈癌Hela细胞系;RT-PCR检测Hela细胞中Survivin mRNA的表达,Western Blot检测Survivin蛋白表达;确定转染成功使干扰组细胞Survivin表达下调后,以流式细胞仪检测细胞凋亡率变化;细胞计数后绘制生长曲线观察细胞增殖能力变化;MTT法检测细胞对抗肿瘤药物的敏感性变化.结果:与对照组相比,两个Survivin基因干扰组的mRNA和蛋白表达抑制率分别为55%,60%和42%,55%,表达均下降(P<0.05);细胞凋亡率分别为19%和22%,较对照组高(P<0.05);生长曲线显示干扰组的细胞增殖能力下降;同一剂量药物作用下,各组细胞对抗肿瘤药物的敏感性均有提高,其中NVB组和PDD组有统计学意义(P<0.05).结论:Survivin SiRNA片段转染进入Hela细胞后,Survivin mRNA和蛋白表达水平均下降,Hela细胞的凋亡率增加,对抗肿瘤药物的敏感性增加.
AIM： To observe the effect of survivin gene repression by RNA interference（RNAi） on the apoptosis and the chemosensitivity of cervical cancer cell line Hela cells. METHODS： The small interfering RNA（siRNA） targeting against survivin was synthesized and transfected into Hela cells with lipofectamine 2000. Survivin expression was detected with the reverse transcription polymerase chain reaction （RT-PCR） and Western Blot. The level of proliferation was evaluated by drawing growth curve after cell counting. Flow cytometry was used to examine cell apoptosis, and the sensitivity to anticancer agents was evaluated by methyl thiazolyl tetrazolium （MTF） assay. RESULTS： Compared with the ceils which were transfected with negative control siRNA （HelaNC）, pure lipofectamine （Hela-lip） and un-transfected cells （Hela）, the expression level of survivin gene mRNA and protein declined evidently in two sets of cells transfected with survivin siRNA （siRNA 1 and siRNA 2 ）, the expression inhibition rates were 55%, 60% and 42%, 55% respectively, the apoptosis rate was increased obviously （ 19% and 22% , P 〈 0.05 ） and the level of proliferation were declined. At the same dose of anticancer agents, the sensitivity of cells in two interferential groups was increased, which was significant in NVB and PDD groups. CONCLUSION： Survivin siRNA can induce apoptosis by knocking down survivin gene expression and enhance chemosensitivity of Hela cells.
出处
《第四军医大学学报》
CAS
北大核心
2008年第20期1843-1846,共4页
Journal of the Fourth Military Medical University
基金
2007年西安市科技创新支撑计划(YF07172)
