siRNA沉默FAK基因表达对乳腺癌MCF-7细胞生物学特性的影响

Effect of silencing FAK gene expression by siRNA on characteristics of human breast cancer MCF-7 cell line

目的:研究RNAi干扰粘着斑激酶(Focal adhesion kinase,FAK)的表达及对人类乳腺癌细胞生物学特性的影响。方法:构建表达FAK基因siRNA的重组质粒FAK-siRNA,在脂质体的介导下转染入人乳腺癌MCF-7细胞,荧光倒置显微镜下观察转染效率,应用RT-PCR和Westernblot检测FAKmRNA和蛋白表达,流式细胞术分析细胞周期分布,体外实验测定肿瘤细胞穿透matrigel的能力。结果:成功转染FAK-siRNA后乳腺癌MCF-7细胞FAK基因的表达与阴性对照组、空白质粒转染组相比FAKmRNA和蛋白表达水平明显降低(P<0.05),细胞增殖指数及侵袭力明显降低(P<0.01)。结论:FAK-siRNA能有效抑制人乳腺癌细胞FAK基因mRNA和蛋白的表达,降低细胞增殖能力及侵袭力。

Objective： To study the effect of RNA interferenc （RNAi） on FAK gene expression and the biological characteristics of human breast cancer MCF-7. Methods： The constructed recombinant plasmid FAK-siRNA was transfected into human breast cancer MCF-7 cell using LipofectamineTM 2000. The transfection efficiency was observed under fluorescence confocal microscope. Expression of FAK mRNA and protein were detected by reverse transcription polymerase chain reaction （RT-PCR） and Western blot. Distribution of cell cycle was assessed by flow cytometry. Cell penetrate matrigel capacity were determined by invitro experiment. Results： FAK-siRNA effectively inhibited FAK mRNA and protein expression （P〈0.05）. The proliferation index and the invasion capability of MCF-7 cells were effectively degraded （P〈0.01）. Conclusion： FAK-siRNA inhibits the expression of FAK mRNA and protein,degrades the proliferation and the invasion capability of human breast cancer cell.