FAK阻断对鼻咽癌细胞增殖、凋亡的影响

Inhibitory effects of FAK on cellular proliferation and apoptosis of nasopharyngeal carcinoma cells

目的:研究RNA干扰(RNAi)抑制黏附斑激酶(FAK)基因表达后对鼻咽癌HNE-1细胞株增殖和凋亡的影响.方法:利用免疫组化法检测鼻咽癌HNE-1细胞株中FAK基因的表达.根据基因库上的FAKmRNA序列,设计合成针对FAK的小干扰RNA(siRNA)并转染HNE-1细胞,收集细胞总RNA和总蛋白,采用RT-PCR,WesternBlot等方法检测细胞中FAK基因表达的变化;利用MTT法检测细胞的生长增殖;流式细胞术观察其对细胞周期及凋亡的影响.结果:鼻咽癌HNE-1细胞株中FAK基因呈阳性表达.针对FAK的siRNA使鼻咽癌HNE-1细胞株FAKmRNA和蛋白表达水平分别下调71.4%,59.8%;MTT法检测显示,细胞的生长增殖受到明显抑制,抑制率可达36.8%;转染后48h,鼻咽癌HNE-1细胞G0/G1期细胞量增加,而S期及G2-M期的细胞减少,同时凋亡率增加.结论:干扰鼻咽癌HNE-1细胞内FAK基因的表达,可抑制肿瘤细胞的生长增殖,使细胞周期重新分布并促进凋亡.

AIM：To investigate the inhibition effects of FAK（focal adhesion kinase）with RNA interference（RNAi）on cellular proliferation and apoptosis of human nasopharyngeal carcinoma（NPC）HNE-1 cells.METHODS：The expression of FAK in HNE-1 cells was detected by immunohistochemical staining.Accordingto FAK mRNA sequence at the GenBank,the siRNA againstFAK were designed and transfected into HNE-1 cells.The expression levels of FAK were confirmed by RT-PCR and Western Blot after the total RNA and protein were collected.The proliferation of HNE-1 cells were determined by MTT assay and the cell cycleand apoptosis were examined by flow cytometry（FCM）.RESULTS：The expression of FAK in HNE-1 cells was positive.The siRNA against FAK downregulated the expression level of mRNA and protein dramatically in HNE-1 cell line,with a down-regulation of 71.4% and 59.8%.The result of MTT indicated that the cancer cellular proliferation was restrained remarkably and the inhibitionratio was 36.8%.At 48 h after the transfection,the number of HNE-1 cells in the G0/G1 phase was increased,that in the S and G2-M phases were depressed.Meanwhile,the apoptosis was increased.CONCLUSION：To interfere expression of FAK in HNE-1 cells,HNE-1 cell proliferation can be inhibited,the cell cycle was induced to redistribute,and so was the cell apoptosis.