RAMP1-siRNA对CGRP促MG-63细胞增殖作用影响的实验研究  

Experimental study on the effect of RAMP1-siRNA on CGRP-induced proliferation of MG-63 cells

作　　者：蒋章[1] 张慧宇[1] 张纲[1] 冯小倩[1] 孙嵩[1] 谭颖徽[1] 

机构地区：[1]第三军医大学附属新桥医院口腔科,重庆400037

出　　处：《实用口腔医学杂志》2015年第3期339-342,共4页Journal of Practical Stomatology

基　　金：国家自然科学基金(编号:81371110;81271098)

摘　　要：目的:探讨阻断降钙素基因相关肽(CGRP)受体活性修饰蛋白(RAMP1)在MG-63细胞中表达对促成骨样MG-63细胞增殖作用的影响。方法:体外转录合成法合成及筛选RAMP1 siRNA,传代培养的MG-63细胞分为:RAMP1 siRNA干扰组、空载体组、空白对照组。实时聚合酶链反应、免疫荧光法分别检测降钙素受体样受体(CRLR)、受体组分蛋白(RCP)在MG-63细胞中mRNA表达及膜上的分布变化。结果:转染siRNA后MG-63细胞RAMP1、CRLR mRNA和荧光强度明显下调(P<0.05),RAMP1干扰组RCP mRNA的表达及荧光强度高于其它2组(P<0.05)。RAMP1 siRNA干扰后,MG-63细胞的增殖被抑制(P<0.05)。结论:转染RAMP1 siRNA降低了MG-63细胞CRLR的表达,抑制了MG-63细胞的增殖能力。Objective：To investigate the effect of blocking the expression of receptor activity modifying protein 1 （RAMP1 ）on calcito-nin gene-related peptide（CGRP）-induced MG-63 cell proliferation.Methods：RAMP1 siRNA was synthesized and screened by tran-scription in vitro.The subcultured MG-63 cells were divided into the following groups：RAMP1 siRNA interference group,empty vector group and blank control group.The mRNA expression and the membrane distribution changes of the calcitonin receptor-like receptor （CRLR）and the receptor component protein （RCP）in MG-63 cells were examined by real-time PCR and immunofluorescence method respectively.Results：RAMP1 and CRLR mRNA and the fluorescence intensity of MG-63 cells decreased after transfection by RAMP1 siRNA（P〈0.05）.In RAMP1 interference group,the expression of RCP mRNA and the fluorescence intensity were higher than those in the other two groups（P ＆lt;0.05）.After RAMP1 siRNA interference,the proliferation of MG-63 cells was inhibited（P〈0.05）. Conclusion：RAMP1 siRNA transfection may reduce CRLR expression and inhibite the proliferation of MG-63 cell.

关 键 词：受体活性修饰蛋白(RAMP1) 降钙素基因相关肽(CGRP) MG-63 增殖 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]