靶向FAK的siRNA对喉癌细胞增殖及侵袭能力的抑制作用  

Effect of focal adhesion kinase gene silencing on proliferation and invasion of Hep-2 cells

作　　者：张潜英[1] 叶琳[1] 余晓燕[1] 沈娜[1] 邓碧[1] 吴晓松[1] 陈鸿雁[1] 

机构地区：[1]重庆医科大学附属第一医院耳鼻咽喉科,重庆400016

出　　处：《第三军医大学学报》2009年第6期526-529,共4页Journal of Third Military Medical University

基　　金：重庆市卫生局资助项目(07-1-021)~~

摘　　要：目的运用RNA干扰(RNA interference,RNAi)技术阻断喉癌Hep-2细胞株中FAK基因的表达,探讨FAK基因沉默后对Hep-2细胞株产生的影响。方法针对FAK基因,构建2条干扰重组质粒FAK-pGenesil-CH1和FAK-pGen-esil-CH2,1条阴性对照重组质粒FAK-pGenesil-HK;在脂质体的介导下转染喉癌Hep-2细胞株,用RT-PCR和Western blot分析FAK mRNA及蛋白的表达;MTT法、流式细胞技术及体外侵袭实验测定干扰重组质粒对Hep-2细胞株的增殖、侵袭能力的影响。结果重组质粒能有效地阻断Hep-2细胞株中FAK基因在mRNA和蛋白水平上的表达(P<0.05);重组质粒转染Hep-2细胞后,细胞增殖抑制率分别为58.34%、52.78%,与对照组比较差异有统计学意义(P<0.05);干扰组G0～G1期细胞增多,S期细胞数量明显减少(P<0.05);FAK-pGenesil-CH1、FAK-pGenesil-CH2作用Hep-2细胞株48h后,Hep-2细胞穿过Matrigel膜的个数分别为(56.0±6.7)、(51.0±5.1),与阴性对照组(152.0±9.4)、正常对照组(139.0±8.1)比较有统计学差异(P<0.05)。结论干扰重组质粒能有效地抑制转染Hep-2细胞株的FAKmRNA及蛋白的表达,并能抑制Hep-2细胞株的增殖及侵袭能力。Objective To observe the effects of RNAi-mediated focal adhesion kinase （FAK） gene silencing on laryngeal carcinoma Hep-2 cells. Methods Recombinant plasmids generating short hairpin RNA were constructed, including two intefferential sequences FAK-pGenesil-CH1, FAK-pGenesil-CH2 and one negative control sequence FAK-pGenesil-HK. Hep-2 cells were transfected with recombinant plasmids. FAK mRNA and protein were analyzed respectively by RT-PCR and Western blotting. Cell proliferation, cell cycle and invasion of Hep-2 cells were observed by MTT assay, flow cytometry and transwell invasion assay, respectively. Results Recombinant plasmids of FAK-pGenesil-CH1 and FAK-pGenesil-CH2 downregulated the expressions of FAK mRNA and protein （ P 〈 0. 05 ）. The proliferation inhibition ratio in Hep-2 cells reached 58.34% and 52.78% 48 h after transfection of recombinant plasmids of FAK-pGenesil-CH1 and FAK-pGenesil-CH2, while that was 8.34% in the control （P 〈 0.05 ）. More cells arrested in G0/G1 phase in the transfected groups than controls. The number of FAK-pGenesil-CH1 transfected Hep-2 cells, and FAK-pGenesil-CH2 transfected Hep-2 cells through Matrigel membrane was （56.0 ±6.7） and （51.0 ±5.1 ）, respectively, significantly less than that in control groups （P 〈 0.05 ）. Conclusion The constructed recombinant plasmids of FAK-pGenesil-CH1 and FAK-pGenesil-CH2 can suppress the expression of FAK mRNA and protein in Hep-2 cells, and inhibit the cloning and invasion of Hep-2 cells.

关 键 词：FAK SIRNA HEP-2细胞株 增殖 侵袭 

分 类 号：R73-37[医药卫生—肿瘤] R730.54[医药卫生—临床医学]