人表皮生长因子受体显性负性突变体诱导胃癌细胞发生G0／G1期阻滞  

Dominant negative human epidermal growth factor receptor induces G_0/G_1 arrest in human gastric cancer cells

作　　者：廖刚[1] 王子卫[1] 张能[2,3] 董浦江[4] 汤为学[5] 

机构地区：[1]重庆医科大学附属第一医院胃肠外科,重庆400016 [2]重庆医科大学 [3]云南省第一人民医院普通外科,云南昆明650032 [4]重庆医科大学附属第一医院实验研究中心,重庆400016 [5]重庆医科大学基础医学院病理生理学教研室,重庆400016

出　　处：《中国病理生理杂志》2013年第3期430-435,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30972872)

摘　　要：目的:探讨人表皮生长因子受体显性负性突变体(dominant negative epidermal growth factor recep-tor,DNEGFR)对胃癌细胞细胞周期的影响及其分子机制。方法:选用2株人胃癌细胞,分为如下6组:SGC-7901细胞未转染组(US组)、SGC-7901细胞pEGFP-N1质粒转染组(ES组)、SGC-7901细胞pEGFPN1-DNEGFR质粒转染组(DS组)、NCI-N87细胞未转染组(UN组)、NCI-N87细胞pEGFP-N1质粒转染组(EN组)和NCI-N87细胞pEGF-PN1-DNEGFR质粒转染组(DN组)。采用流式细胞术检测细胞周期,Western blotting检测细胞周期素依赖性蛋白激酶2(CDK2)、cyclin D1、Ser9位点磷酸化糖原合成酶激酶3β[p-GSK-3β(Ser9)]、p21和p27蛋白水平。结果:转染pEGFPN1-DNEGFR质粒的人胃癌细胞株出现G0/G1期阻滞,CDK2、cyclin D1和p-GSK-3β(Ser9)蛋白水平降低,p21和p27蛋白水平则升高。结论:DNEGFR通过激活GSK-3β使cyclin D1蛋白水平降低,并降低CDK2蛋白水平,上调p21和p27蛋白水平,最终导致胃癌细胞发生G0/G1期阻滞。这一结果将为胃癌生物治疗研究提供新思路。AIM： To explore the effect of dominant negative epidermal growth factor receptor （DNEGFR） on the cell cycle of human gastric cancer cells. METHODS ： Two human gastric cancer cell lines were used in the study. The cells were divided into 6 groups, including untreated SGC-7901 cells （US group）, SGC-790! cells stably transfected with pEGFP-N1 （ES group）, SGC-7901 cells stably transfected with pEGFPN1-DNEGFR （DS group）, untreated NCI-N87 cells （UN group）, NCI-N87 cells stably transfected with pEGFP-NI （EN group）, and NCI-N87 cells stably transfected with pEGFPN1-DNEGFR （ DN group）. The cell cycle was determined by flow cytometry. The protein levels of cyclin-de- pendent kinase 2 （CDK2）, cyclin D1, phosphorylated glycogen synthase kinase 3 beta at Ser9 [ p-GSK-313 （Ser9） ], p21 and p27 were detected by Western blotting. RESULTS： Transfection of the human gastric cancer cells with pEGFPN1- DNEGFR led to Go/G~ arrest, and down-regulated CDK2, cyclin D1, p-GSK-313 （Ser9） and up-regulated p21 and p27 as well. CONCLUSION： DNEGFR down-regulates cyclin D1 by activating GSK-313, down-regulates CDK2, and up-regu- lates p21 and p27, which induce G0/G1 arrest in human gastric cancer cells in the end.

关 键 词：表皮生长因子受体 显性负性突变体 胃肿瘤 细胞周期 

分 类 号：R735.2[医药卫生—肿瘤]