Xiong Huihua, Yu Shiying, Hu Guangyuan, Zhuang Liang
Department of Oncology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
J Huazhong Univ Sci Technolog Med Sci. 2006;26(3):305-7. doi: 10.1007/BF02829558.
In order to investigate the effects of short hairpin RNA (shRNA) on the expression of Survivin, cell cycle and cell proliferation in MCF-7 cells, using a pEGFP vector which contained a U6 promoter shRNA plasmid targeted against survivin was constructed and transfected into MCF-7 cells. The change of the expression of Survivin and cell proliferation rates were detected by semiquantitative reverse transcription polymerase chain reaction (RT-PCR) and MTT methods respectively. The change of cell cycle after transfection was analyzed by flow cytometry. The results indicated that the recombinant plasmid containing Survivin shRNA was constructed successfully, which could suppress the expression of Survivin at mRNA and protein level. The growth of MCF-7 cells was arrested in G1 phase of the cell cycle and the proliferation activity was suppressed after transfection. It was concluded that Survivin shRNA plasmid could knock down the expression of Survivin in MCF-7 cells specifically. In addition, Survivin shRNA plasmid could lead to G1 arrest and inhibit the proliferation of MCF-7 cells, which suggested that Survivin shRNA might be used as a new therapeutic method for breast cancer.
为了研究短发夹RNA(shRNA)对MCF-7细胞中Survivin表达、细胞周期和细胞增殖的影响,构建了一种含有针对Survivin的U6启动子shRNA质粒的pEGFP载体,并将其转染到MCF-7细胞中。分别采用半定量逆转录聚合酶链反应(RT-PCR)和MTT法检测Survivin表达变化和细胞增殖率。通过流式细胞术分析转染后细胞周期的变化。结果表明,成功构建了含Survivin shRNA的重组质粒,其可在mRNA和蛋白水平抑制Survivin的表达。转染后MCF-7细胞的生长停滞在细胞周期的G1期,增殖活性受到抑制。结论是,Survivin shRNA质粒可特异性下调MCF-7细胞中Survivin的表达。此外,Survivin shRNA质粒可导致G1期阻滞并抑制MCF-7细胞的增殖,这表明Survivin shRNA可能作为一种新的乳腺癌治疗方法。