摘要
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 .0 73(P <0 .0 0 5 ) ,增殖抑制率达到了94 .5 % ,凋亡率为 76 .5 % (P <0 .0 1) ,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。
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.
出处
《重庆医科大学学报》
CAS
CSCD
2004年第5期567-571,共5页
Journal of Chongqing Medical University
基金
国家自然科学基金资助项目 (编号 :3 0 0 70 2 3 0 )
关键词
舌鳞癌
RNA干扰
转录后沉默(PTGS)
联合转染
Tongue carcinoma
RNA interfering
Post transcriptional gene silencing(PTGS)
Combined transfection
