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
沈娜 1张潜英 1余晓燕 1叶琳 1陈鸿雁1
作者信息
- 1. 重庆医科大学附属第一医院耳鼻咽喉科,重庆,400016
- 折叠
摘要
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FAK mRNA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,Western Blot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FAK mRNA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48 h,鼻咽癌HNE-1细胞G0/GI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.
Abstract
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. According to FAK mRNA sequence at the GenBank, the siRNA against FAK 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 cycle and 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 inhibition ratio 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.
关键词
小干扰RNA/FAK/增殖/凋亡Key words
siRNA/FAK/proliferation/apoptosis引用本文复制引用
出版年
2009
NETL
NSTL
维普
万方数据