肝细胞黏附分子调节蛋白激酶C ε对肾癌786-O细胞增殖和迁移的影响  

Effect of hepatocyte cell adhesion molecule regulating protein kinase C ε on proliferation and migration of renal carcinoma 786-O cells

作　　者：谭兵[1] 徐向东[1] 全真 姜小良[1] 杜红飞[2] 罗春丽[2] 吴小候[1] 

机构地区：[1]重庆医科大学附属第一医院泌尿外科,重庆400016 [2]重庆医科大学检验医学院临床检验诊断学教育部重点实验室,重庆400016

出　　处：《重庆医科大学学报》2014年第6期748-752,共5页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(编号:81072086)

摘　　要：目的:研究肝细胞黏附分子(hepatocyte cell adhesion molecule,HepaCAM)对肾癌786-O细胞增殖及迁移的影响。方法:以786-O为目的细胞,通过感染HepaCAM腺病毒(Ad-GFP-HepaCAM)和空载病毒(Ad-GFP),Western blot检测各处理组蛋白激酶Cε(protein kinase Cε,PKCε)膜蛋白、浆蛋白和总蛋白的变化,Ad-GFP-HepaCAM和10μmol/L PKCε特异性转位抑制剂(εV1-2)处理24 h后基质金属蛋白酶(matrix metalloproteinase,MMP)-9和细胞周期蛋白cyclinD1的变化;平板克隆和划痕实验检测Ad-GFP-HepaCAM及εV1-2对786-O增殖和迁移的影响。结果:感染Ad-GFP-HepaCAM后与对照组比较,PKCε膜蛋白降低(F=82.724,P=0.000;P=0.000)而浆蛋白升高(F=537.006,P=0.000;P=0.000),总蛋白无统计学差异(F=0.548,P=0.605;P=0.805)。Ad-GFP-HepaCAM和εV1-2处理后,MMP-9降低(F=33.109,P=0.001,P=0.000;F=96.868,P=0.000,P=0.000),cyclinD1降低(F=562.626,P=0.000,P=0.000;F=159.433,P=0.000,P=0.000)。Ad-GFP-HepaCAM组比εV1-2组克隆形成率降低(F=259.918,P=0.000;P=0.000),迁移率降低(F=550.169,P=0.000;P=0.000)。结论:HepaCAM可能通过阻止PKCε从胞浆转位到胞膜而部分失活来抑制786-O细胞增殖及迁移。Objective：To investigate the effect of re-expression of hepatocyte cell adhesion molecule（HepaCAM）on the proliferation and migration ability of renal carcinoma 786-O cells. Methods：Protein kinase C ε（PKCε）membrane protein level,PKCε cytosolic protein level and total protein level were determined by Western blot after 786-O cell being infected with Ad-GFP-HepaCAM and Ad-GFP. Matrix metalloproteinase（MMP）-9 and cyclinD1 protein levels were also detected by Western blot after being treated with Ad-GFP-HepaCAM and εV1-2（10 μmol/L,24 h）. The proliferation ability and migration ability of 786-O cells were analyzed by colony formation assay and wound healing assay,respectively. Results：When compared with those of control group,PKCε membrane protein level was decreased（F=82.724,P=0.000;P=0.000）and cytosolic protein level was increased（F=537.006,P=0.000;P=0.000）,but PKCε total protein level was not changed（F=0.548,P=0.605;P=0.805）after being infected with Ad-GFP-HepaCAM. After exposure to Ad-GFP-HepaCAM and εV1-2,MMP-9 protein level was down-regulated（F=33.109,P=0.001,P=0.000;F=96.868,P=0.000,P=0.000）and cyclinD1 protein level was also down-regulated（F=562.626,P=0.000,P=0.000;F=159.433,P=0.000,P=0.000）when compared with those of control group. The colony formation rate was decreased（F=259.918,P=0.000;P=0.000）and migration ratio was also reduced（F=550.169,P=0.000;P=0.000）in Ad-GFP-HepaCAM group compared with those of εV1-2 group. Conclusion：HepaCAM could inhibit 786-O cell proliferation and migration via blocking PKCε translocating from cytoplasm to membrane and inactivating PKCε partially.

关 键 词：肝细胞黏附分子 蛋白激酶CΕ 肾癌 增殖 迁移 

分 类 号：R737.11[医药卫生—肿瘤] R730.23[医药卫生—临床医学]