Department of Obstetrics and Gynecology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, China.
Mol Biol Rep. 2012 Dec;39(12):10665-70. doi: 10.1007/s11033-012-1956-6. Epub 2012 Oct 19.
The study investigates the effect of FAK, DLC-1 on OVCAR-3 proliferation. FAK gene siRNA vector recombinant plasmid was constructed using RNA interference technique. FAK gene-transfected OVCAR-3 cells, OVCAR-3 cells with DLC-1 gene expression, and OVCAR-3 cells with simultaneous expression of DLC-1 and FAK genes were obtained using gene transfection technology. In addition, siRNA control group and blank control were also given. Effect of FAK, DLC-1 gene expression on OVCAR-3 proliferation was examined by FCM and Cell Counting Kit-8 (CCK-8) methods. Results showed that DLC-1 gene high expression and FAK gene silencing, single silencing FAK gene, and single DLC-1 gene high expression in OVCAR-3 cells may decrease S and G2/M phase proportion of the cell cycle. Moreover, DLC-1 gene high expression and FAK gene silencing in OVCAR-3 cells can display the most significant effect. This confirmed that DLC-1 gene high expression and FAK gene silencing may significantly inhibit the OVCAR-3 cells proliferation. CCK-8 analysis showed that silence FAK gene expression or/and increasing DLC-1 gene expression may decrease OVCAR-3 growth rate. Moreover, simultaneous silence the expression of FAK gene and high expression of DLC-1 gene can display the most significant effect on OVCAR-3 growth. It can be concluded that downregulation of FAK gene expression or/and upregulation of DLC-1 gene expression can all inhibit the OVCAR-3 growth. Moreover, DLC-1 gene expression and FAK gene silencing can display the most marked inhibitory effect on the OVCAR-3 growth.
本研究探讨了 FAK、DLC-1 对 OVCAR-3 增殖的影响。采用 RNA 干扰技术构建 FAK 基因 siRNA 载体重组质粒，采用基因转染技术获得 FAK 基因转染 OVCAR-3 细胞、DLC-1 基因表达的 OVCAR-3 细胞以及同时表达 DLC-1 和 FAK 基因的 OVCAR-3 细胞，同时设立 siRNA 对照组和空白对照组。采用 FCM 和细胞计数试剂盒（CCK-8）方法检测 FAK、DLC-1 基因表达对 OVCAR-3 增殖的影响。结果显示，DLC-1 基因高表达和 FAK 基因沉默、单独沉默 FAK 基因以及单独高表达 DLC-1 基因均可降低 OVCAR-3 细胞周期 S 和 G2/M 期比例。而且，DLC-1 基因高表达和 FAK 基因沉默在 OVCAR-3 细胞中作用最显著。这证实了 DLC-1 基因高表达和 FAK 基因沉默可显著抑制 OVCAR-3 细胞增殖。CCK-8 分析表明，沉默 FAK 基因表达或/和增加 DLC-1 基因表达均可降低 OVCAR-3 生长速度。而且，同时沉默 FAK 基因表达和高表达 DLC-1 基因对 OVCAR-3 生长的抑制作用最显著。由此可以得出结论，下调 FAK 基因表达或/和上调 DLC-1 基因表达均可抑制 OVCAR-3 生长，而且 DLC-1 基因表达和 FAK 基因沉默对 OVCAR-3 生长的抑制作用最显著。
