RNAi沉默c-erb、B-2、c-raf-1表达对人舌鳞癌Tca8113细胞的干预作用

目的:探讨应用RNA干扰技术沉默c-erbB-2、c-raf-1基因表达,研究其对人舌鳞癌Tca8113细胞增殖的抑制作用及其分子机制.方法:实验分成3组:空白对照组、正常对照组、c-erbB-2、c-raf-1dsRNA联合干扰组.脂质体转染后不同时间分别进行MTT试验、电镜检测、RT-PCR,以观察不同条件处理后各组细胞的增殖、凋亡、细胞周期、mRNA及蛋白质表达有无差别.结果:c-erbB-2、c-raf-1dsRNA联合干扰组,转染72h的OD值为0.073(P＜0.005),增殖抑制率达到了94.5%,凋亡率为76.5%(P＜0.01),明显地下调c-erbB-2与c-raf-1基因mRNA及其蛋白质的表达水平.结论:在舌癌Tca8113细胞株增殖的抑制作用研究中,c-erbB-2、c-raf-1dsRNA联合干扰具有显著沉默c-erbB-2、c-raf-1基因表达.

RNA interference c - erbB - 2 and c - raf - 1 genes expression combined transfection in the Tca8113 cell lines

Objective:To investigate the proliferating inhibition and the mechanisms of dsRNA interference c-erbB-2 and c-raf-1 genes expression combined transfection in the human tongue carcinoma Tca8113 cell lines.Methods:There were 3 groups in our study,control group,normal control group and RNAi experimental group.At different time after liposome-mediated transfection,the cell proliferation,apoptosis,mRNA level,protein expressing level and cell cycle were observed by MTT,RT-PCR and electronic microscope.Results:According to the results of RNAi experimental group,the OD-value were 0.073( P <0.005)transfected after 72h;the inhibitory rate of proliferation were 94.5%;the apoptosis rate were 76.5% ( P <0.01 ) and the level of mRNA and protein were decreased statistically in both c-erbB-2 and c-raf-1.Conclusion:These results suggest that c-erbB-2 and c-raf-1 dsRNA effects of silencing c-erbB-2 and c-raf-1 genes expression combined transfection of tongue carcinoma.