FAK siRNA重组质粒的构建及其对胶质瘤U251细胞增殖及侵袭能力的抑制作用

目的 探讨应用RNA干扰技术沉默黏着斑激酶（FAK）基因表达对人脑胶质瘤U251细胞增殖和侵袭能力的影响。方法 将构建成功的FAK siRNA重组质粒转染至U251细胞，合成与FAK基因序列无关的siRNA作为阴性对照，以野生型U251细胞作为空白对照组。运用PCR和免疫印迹法观察FAK mRNA和蛋白表达情况，实时细胞分析仪检测观察细胞增殖情况，Transwell小室侵袭模型研究细胞侵袭能力。结果 重组质粒pBSilence1.1-FAK构建成功；与空白对照组和阴性对照组相比，干扰组细胞增殖减慢（P <0.05）；Transwell小室体外侵袭结果 显示，干扰组穿膜细胞数仅为（17.1±1.0），明显低于空白对照组（68.2±4.5；P <0.05）和阴性对照组（67.0±2.3；P <0.05）。结论 沉默FAK基因表达可有效抑制人胶质瘤U251细胞的增殖和侵袭能力。

Construction of siRNA recombinant plasmid targeting focal adhesion kinase gene and its effect on the proliferation and invasiveness of human glioma U251cells

Objective To explore the effect of silencing focal adhesion kinase (FAK) gene on the proliferation and invasiveness of human glioma U251 cells. Methods U251 cells were divided into 3 groups, i.e. blank group, negative control group and experiment group. The FAK small interference RNA (siRNA) recombinant plasmid was transfected into human glioma U251 cells in the experimental group. The expressions of FAK mRNA and protein were detected by real time PCR and Western-blot respectively, and the U251 cells proliferative and invasive capabilities were measured respectively by RT cell analyzer (RTCA) and Transwell chamber invasion assay. Results The pBSilence1.1-FAK recombinant plasmid was successfully constructed. RTCA Results showed that U251 cell proliferative capability was significantly lower in the experimental group than those in the blank group and the negative control group (P <0.05). Transwell chamber invasion assay showed that the number of U251 cells of transwell chamber [(17.137±1.012) cells] was significantly fewer in the experimental group than those in the blank group [(68.218±4.547) cells] and the negative control group [(67.034±2.272) cells] (P <0.05). Conclusion Silencing FAK gene can significantly inhibit proliferation and invasveness of human glioma U251 cells in vitro.