小分子干扰RNA抑制人皮肤鳞状细胞癌A431细胞株中生存素的实验研究  

Inhibitory effect of siRNA targeting survivin on the growth of human squamous carcinoma cells

作　　者：宋琳毅[1] 江阳[2] 

机构地区：[1]苏州大学附属第一医院皮肤科,苏州215006 [2]上海交通大学医学院九龙医院,苏州215021

出　　处：《现代免疫学》2009年第4期283-287,共5页Current Immunology

摘　　要：探讨小干扰RNA(siRNA)对人皮肤鳞状细胞癌A431细胞株生存素基因表达的抑制作用及对细胞凋亡、增殖和细胞周期的影响。将已构建的靶向生存素的siRNA表达质粒采用脂质体法转染A431细胞株。应用RT-PCR检测A431细胞生存素mRNA的表达,Western blot检测生存素蛋白的表达,流式细胞术检测Annexin标记的凋亡细胞和分析细胞周期,噻唑蓝(MTT)法分析细胞增殖。结果显示,与未处理的A431细胞株和转染空载体的A431细胞株比较,转染siRNA表达质粒可以明显抑制生存素基因在转录和翻译上的表达;siRNA表达质粒组的凋亡率明显增加达到21.3%;24、48、72和96h的细胞增殖抑制率也明显增加,并随着时间抑制效率变高;细胞周期分析发现抑制生存素表达使得A431细胞株出现明显的G2/M期阻滞现象。因此,靶向生存素的siRNA表达质粒可以特异性抑制生存素的表达,显著抑制肿瘤细胞增殖并诱导细胞凋亡。靶向生存素的siRNA表达质粒为鳞状细胞癌的基因治疗提供了一种新思路。To investigate the inhibitory effect of siRNA targeting survivin on apoptosis and proliferation of human squamous carcinoma cells, the constructed siRNA expression vector targeting survivin pRNAT-H1.1 was transfected with lipofectamine 2000 liposome. The mRNA expression of surviving in A431 cells was detected by RT-PCR, and the expression of surviving protein was detected by Western blotting. The Annexin V iabeled apoptotic cells and the cell cycle of A431. Cells were analyzed by flow eytometry and the proliferative activities of A431 cells were measured by MTT method. As demonstrated by RT PCR analysis, the transfected siRNA expression plasmid could inhibit significantly the expression the survivin gene in tran scription and translation level in comparison with the un-treated A431 cells. The apoptotic cell rate of the siRNA expression plasmid group increased markedly up to 21. 3%, and the rates of inhibition in cell proliferation were also significantly in creased in a time-dependent manner. In addition, analysis of cell cycle demonstrated a marked blockage of the G2/M stage of the A431 cells treated with siRNA expression plasmid. It is concluded that siRNA against survivin gene can induce apoptosis and inhibit proliferation of human squamous carcinoma cells, thus it may provide a rational approach in gene therapy for human squamous carcinoma.

关 键 词：生存素 RNA干扰 鳞状细胞癌 凋亡 增殖 

分 类 号：R392.4[医药卫生—免疫学]