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

发布时间：2018-05-25 14:18

本文选题：DEPDC + 鼻咽癌　；参考：《中国细胞生物学学报》2014年03期

【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。
[Abstract]:In order to investigate the effect of silencing DEPDC1 gene expression on the growth and cell cycle of nasopharyngeal carcinoma (NPC) cell line HNE-1, small interfering RNA(small interfering RNAs targeting DEPDC1 were synthesized and transfected into human nasopharyngeal carcinoma HNE-1 cells. After transfection, the expression of DEPDC1, cell cycle, growth and proliferation, apoptosis and its possible mechanism were detected by fluorescence quantitative PCR, Western blotting and flow cytometry. The results showed that the expression of DEPDC1 gene in mRNA and protein was significantly decreased after DEPDC1 siRNA transfection, and a large number of cells were blocked in G _ 2 / M phase, growth and proliferation slowed and apoptosis increased. The results of fluorescence quantitative PCR showed that the expression of NF- 魏 B activated A20 gene was up-regulated and the expression of NF- 魏 B regulated tumor related target gene was decreased, including the expression of ICAM-1BCL-2 gene. It is suggested that silencing DEPDC1 gene can affect the cell cycle of HNE-1 cells, inhibit its growth and proliferation, and promote apoptosis, and its mechanism may be related to the inhibition of NF- 魏 B pathway.
【作者单位】：重庆医科大学附属第一医院耳鼻咽喉科;重庆医科大学分子医学与肿瘤研究中心;
【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~~
【分类号】：R739.63

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