siRNA-cyclin D1对乳腺癌MCF-7细胞增殖的抑制作用

siRNA-CYCLIN D1 INHIBIT CELL PROLIFERATION IN BREAST CANCER MCF-7 CELL LINE

研究小干扰RNA(small interfering RNA,siRNA)对乳腺癌MCF-7细胞株cyclin D1表达的抑制及对细胞增殖的影响。化学合成针对cyclin D1基因的siRNA,转染MCF-7细胞株；分别应用荧光定量PCR和免疫印迹测定cyclin D1 mRNA和蛋白的表达,CCK-8测定细胞的增殖活性,流式细胞仪检测细胞周期,软琼脂培养检测细胞克隆形成能力。在实验中,10、50、100 nmol/L siRNA-cyclin D1分别使MCF-7细胞cyclin D1 mRNA表达降低了57．85%、63．22%和68．02%,蛋白表达降低了51．13%、62．09%、77．68%。转染siRNA-cyclin D1后,细胞增殖受到抑制,细胞周期阻滞于G1期,软琼脂克隆形成率降低。结果提示siRNA可以有效抑制MCF-7细胞株中cyclin D1的表达,使细胞周期阻滞于G1期,从而抑制细胞增殖。

To study the inhibitory effect of siRNA on cyclin D1 expression and cell proliferation in breast cancer MCF-7 cell line. The siRNA targeting cyclin D1 was chemically synthesized and transfected into MCF-7 cells by oligofectamine. The expression of cyclin D1 was analyzed by quantitive PCR and Western blot, and the cell growth inhibition was measured with CCK-8 assay. Then, cell cycle of the transfected cells was examined by flow cytometry, and cell colony forming ability was measured by soft-agar colony formation assay. After MCF-7 cells were transfected with 10,50,100nmol/L siRNA,the expression of cyclin D1 mRNA was respectively suppressed with inhibition rates of 57.85%, 63.22% and 68.02%, and the protein expression was suppressed with inhibition rates of 51.13%, 62.09% and 77.68% respectively. The proliferation of MCF-7 cells was inhibited after transfection with siRNA-cyclin D1, which caused cell cycle arrest at G1 phase and showed less colony forming ability in the breast cancer cell line MCF-7. These results indicate that siRNA-cyclin D1 could be a powerful anti-proliferative tool in breast cancer gene therapy.