siRNA封闭BRCAA1基因对乳腺癌细胞MCF-7增殖和Rb基因表达的影响  

Effect of blocking BRCAA1 gene with siRNA on proliferation of MCF-7 cells and expression of Rb gene

作　　者：杨浩[1] 崔大祥[1] 李清[1] 黄拓[1] 高峰[1] 贺蓉[1] 潘碧峰[1] 邵君[1] 尤晓钢[1] 刘风涛[1] 

机构地区：[1]上海交通大学微纳科学和技术研究院

出　　处：《中国肿瘤生物治疗杂志》2006年第3期181-184,共4页Chinese Journal of Cancer Biotherapy

基　　金：国家重点基础研究发展(973)项目(2005CB724300-G);国家自然科学基金(39900177;30471599)

摘　　要：目的:研究siRNA封闭乳腺癌抗原相关基因1(BRCAA1)对乳腺癌细胞MCF-7增殖和Rb基因表达的影响。方法:采用RNAi技术对乳腺癌细胞MCF-7细胞BRCAA1基因进行特异性抑制,用阳离子脂质体与化学合成的Pre-designedan-ti-BRCAA1 siRNA构建转染复合体,反转染MCF-7细胞株48h后提取总RNA,分为未处理(NT)组、阴性对照组、阳性对照组和BRCAA1组,经反转录荧光实时PCR检测BRCAA1和Rb基因mRNA表达情况;检测细胞增殖抑制率。结果:与阴性对照组相比,siRNA转染MCF-7细胞后实验组BRCAA1基因mRNA水平降低了42.3%;Rb基因表达较之阴性对照组上升了11.1%;实验组MCF-7细胞增殖抑制率为(81.9±6.1)%,抑制作用明显强于对照组(P<0.05)。结论:BRCAA1基因的封闭明显抑制了MCF-7细胞的增殖,BRCAA1基因与Rb基因可能存在有某种相互拮抗的作用。Objective： To investigate the effect of blocking BRCAA1 gene expression with siRNA on the proliferation of tumor cell line MCF-7 and Rb gene expression. Methods： RNAi was employed to specifically knock down BRCAA1 expression. MCF-7 cells were transfected with complexes constructed with lipids and chemically synthesized Pre-designed anti-BRCAA1 siRNAs. The total RNA was isolated and reversely transcribed after 48 h. The expressions of BRCAA1 and Rb mRNA were determined by Real-Time PCR. Results： Compared with negative control, transfected MCF-7 cells had a 42.3% decrease in expression of BRCAA1 mRNA and an 11.1% increase in Rb mRNA expression. The inhibitory rate of MCF-7 cells proliferation was （81.6 ± 6. 1 ）%. Conclusion： There may be some antagonistic effect between BRCAA1 gene and Rb gene in proliferation of tumor ceils, which provides a potential target for anti-tumor gene therapy.

关 键 词：SIRNA BRCAA1基因 乳腺癌 RB基因 

分 类 号：R730[医药卫生—肿瘤]