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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 siRNA-containing breast cancer antigen-related gene 1 (BRCAA1) on the proliferation and Rb gene expression of breast cancer cell line MCF-7. METHODS: The BRCAA1 gene was specifically inhibited in human breast cancer cell line MCF-7 by using RNAi technique. The transfection complex was constructed by using cationic liposomes and chemically synthesized Pre-designedan-ti-BRCAA1 siRNA, and then transfected into MCF-7 cells After 48h, the total RNA was extracted and divided into untreated (NT) group, negative control group, positive control group and BRCAA1 group. The expression of BRCAA1 and Rb mRNA was detected by reverse transcription real-time PCR. Results: Compared with the negative control group, the expression of BRCAA1 mRNA decreased by 42.3% and the expression of Rb gene increased by 11.1% in MCF-7 cells compared with the negative control group. The proliferation inhibition rate of MCF-7 cells in experimental group (81.9 ± 6.1)%, the inhibitory effect was significantly stronger than the control group (P <0.05). CONCLUSION: Blocking of BRCAA1 gene significantly inhibits the proliferation of MCF-7 cells. There may be some antagonism between BRCAA1 gene and Rb gene.