高血压相关基因hrg-1对血管平滑肌细胞周期蛋白E和P27蛋白表达及细胞增殖的影响  

Overexpression of hrg-1 gene affects the expression of cyclin E,P27 protein and proliferation of VSMCs

作　　者：姜广建[1] 温进坤[1] 韩梅[1] 潘磊[2] 黄秀英[2] 

机构地区：[1]河北医科大学基础医学研究所河北省医学生物技术重点实验室,河北石家庄050017 [2]中国科学院遗传与发育研究所

出　　处：《中国老年学杂志》2004年第10期923-926,共4页Chinese Journal of Gerontology

基　　金：国家自然科学基金 (30 2 70 4 99;90 2 0 80 1 4 );河北省自然科学基金(30 3455)资助项目

摘　　要：目的　研究hrg 1对血管平滑肌细胞 (VSMC)增殖的抑制作用是否与调节细胞周期调节蛋白 (cyclin)表达活性有关。 方法　采用pcD2 hrg 1真核表达质粒转染体外培养的VSMCs,Westernblot、MTT法、流式细胞术观察该基因过表达对细胞周期蛋白E(cyclinE)、P2 7表达及细胞增殖和细胞周期的影响。结果　转染pcD2 hrg 1后 ,与对照组细胞相比 ,G1期细胞比例上升 ,S期细胞比例下降 ,细胞生长速率明显减慢 ;而转染hrg 1反义RNA表达载体的细胞生长速率明显加快。转染hrg 1 2 4h后cyclinE表达开始降低 ,之后呈持续下降趋势 ;至 72h ,其表达量约为对照组的 1 /2。P2 7的表达则呈上升趋势 ,转染hrg 1 2 4h后 ,其表达活性显著增加 (为对照组的 2 6倍 ) ;转染后 48h达高峰 ,为对照组的 3 3倍 ;在转染后72h仍维持在较高水平。结论　hrg 1通过上调P2Objective To study whether the inhibition of VSMC proliferation by hrg-1 is related to the expression activity of cyclins.Methods We infected pcD2-hrg-1 expression plasmids into cultured VSMCs in vitro, and then observed the effect of hrg-1 overexpression on expression of cyclin E, P27, VSMC proliferation and cell cycle. Results The proportion of G 1-phase cells increased after transfecting pcD2-hrg-1 into VSMCs. On the other hand, the proportion of S-phase cells decreased. The cell growth rate was obviously slower than that of control cells. Whereas in the VSMCs infected with antisense hrg-1 plasmids, the cell growth rate was faster than that of control cells. The expression of cyclin E began to decrease 24 hours after transfection, and continued to reduce later. Compared with the cells transfected with control plasmids, it decreased about 50% 72 hours after transfection. Otherwise the expression of P27 trended toward increase. After 24 hours, the level of P27 was about 2.6 times as high as that of control cells, and reached peak (3.3 times of control cells) at 48h after transfection, and then maintained at high level until 72 hours. Conclusions HRG-1 could inhibit VSMC proliferation by up-regulating the expression of P27 and down-regulating the expression of cyclin E.

关 键 词：转染 表达活性 对照组 VSMC 血管平滑肌细胞 细胞增殖 cD2 表达载体 表达量 细胞生长 

分 类 号：R735[医药卫生—肿瘤] R737[医药卫生—临床医学]