沉默DEPDC1基因表达对鼻咽癌细胞系HNE-1生长和细胞周期的影响  

The Effects of DEPDC1 Depletion on Growth and Cell Cycle in Nasopharyngeal Carcinoma Cell Line HNE-1

作　　者：何龙霞[1] 糜燕[1] 张春冬[2] 张莹[2] 蔡伟[2] 王义涛[2] 卜友泉[2] 朱江[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 [2]重庆医科大学分子医学与肿瘤研究中心,重庆400016

出　　处：《中国细胞生物学学报》2014年第3期349-355,共7页Chinese Journal of Cell Biology

基　　金：国家临床重点专科建设项目经费基金卫办医政[2012]649号;重庆市卫生局重点项目(批准号:2013-1-004)资助的课题~~

摘　　要：为探讨沉默DEPDC1基因表达对鼻咽癌细胞系HNE-1生长和细胞周期的影响,该实验设计合成靶向DEPDC1的小分子干扰RNA(small interfering RNA,siRNA)转染人鼻咽癌HNE-1细胞。转染后,采用荧光定量PCR、免疫印迹、MTT及流式细胞术方法检测细胞内DEPDC1的表达量以及细胞周期、生长增殖、凋亡的变化及其可能机制。结果显示,转染DEPDC1 siRNA后,DEPDC1基因在mRNA及蛋白水平的表达量明显降低;大量细胞被阻滞于G2/M期,生长增殖减慢,凋亡增加。荧光定量PCR结果表明,抑制NF-κB激活的A20基因表达量明显上调,受NF-κB调控的肿瘤相关靶基因的表达量下降,包括C-MYC、MMP9、ICAM-1、BCL-2基因。由此说明,沉默DEPDC1基因可以影响HNE-1细胞的周期,抑制其生长增殖,促进凋亡,其机制可能与抑制NF-κB通路有关。To investigate the potential role of DEPDC1 in nasopharyngeal carcinoma, the specific siRNAs against DEPDC1 was designed, synthesized and transiently transfected into the nasopharyngeal carcinoma cell line HNE-1. The results of RT-PCR and Western blot clearly demonstrated that the expression of DEPDC1 was efficiently inhibited at both mRNA and protein levels. FACS （fluorescence activated cell sorter）, MTT and apoptosis assays demonstrated that DEPDC1 depletion resulted into the GE/M arrest, growth retardation and apoptosis. In addition, results of RT-PCR showed that DEPDC1 depletion caused the upregulation of A20 gene, an inhibitor of NF-κB activation, and the downregulation of several NF-κB target genes such as C-MYC, MMPg, ICAM-1 and BCL-2. Taken together, our results suggested that siRNA-mediated DEPDC1 depletion caused cell cycle arrest, led to growth inhibition induced apoptosis in the nasopharyngeal carcinoma cell line HNE-1, which were possibly mediated by the inactivation of NF-κB pathway.

关 键 词：DEPDC1 鼻咽癌 SIRNA 细胞周期 生长 凋亡 

分 类 号：R739.63[医药卫生—肿瘤]