吉林大学学报(医学版)

• 基础研究 • 上一篇    下一篇

RNA干扰COPS3基因对肺癌细胞A549增殖的抑制作用及其机制

曹鹤1,李薇1,钱磊1,王雪梅2,崔久嵬1   

  1. (1.吉林大学第一医院肿瘤中心,吉林 长春 130021;2.吉林省人民医院血液科,吉林 长春 130021)
  • 收稿日期:2013-10-22 出版日期:2014-03-28 发布日期:2014-03-28
  • 通讯作者: 崔久嵬 E-mail:cuijiuwei@vip.qq.com
  • 作者简介:曹鹤(1985-),女,吉林省通化市人,在读医学硕士,主要从事肺部肿瘤和血液病基础及临床研究。
  • 基金资助:

    国家自然科学基金青年基金资助课题(Grant NO.81071920,30901702);教育部科学技术重点项目资助课题(311015);吉林省科技厅中青年领军人才

    创新团队项目资助课题(20111807);吉林大学杰出青年基金资助课题(201005001)

Inhibitory effect of COPS3 gene interferred by RNA on proliferation of lung cancer A549 cells  and its mechanism

CAO He1,LI Wei1,QIAN Lei1,WANG Xue-mei2,CUI Jiu-wei1   

  1. (1.Cancer Center,First Hospital,Jilin University,Changchun 130021,China;2. Department of Hematology,People’s Hospital, Jilin Province,Changchun 130021,China)
  • Received:2013-10-22 Online:2014-03-28 Published:2014-03-28

摘要:

目的:探讨RNA干扰构成型光形态建成同源亚基3(COPS3)基因对肺癌细胞A549增殖的抑作用,初步阐明其作用机制。方法:选取肺腺癌细胞系A549建立

裸小鼠成瘤实验模型,将40只小鼠随机分为实验组(移植感染si-COPS3的A549细胞)和对照组(移植感染si-CTRL的A549细胞),每组20只,检测实验组和

对照组裸小鼠体内的成瘤体积、质量和成瘤速率。应用Western blotting法检测抑制COPS3基因表达后细胞周期相关蛋白P21、CDK2、CDK4、cyclinB1

和cyclinD1表达水平。结果:实验组裸小鼠体内成瘤体积和质量显著小于对照组(P<0.05),成瘤速率较对照组明显下降。与对照组比较,实验组小

鼠细胞P21蛋白表达水平明显上调(P<0.05),cyclin B1和CDK4表达水平下调(P<0.05),而cyclin D1与CDK2表达水平无明显变化(P=0.384,

P=0.605)。结论:COPS3基因表达受抑制使肺癌细胞增殖能力明显下降,其作用机制与P21表达上调、cyclin B1和CDK4表达下调有关联。

关键词: 构成型光形态建成同源亚基3, RNA干扰, 肺肿瘤, 慢病毒载体

Abstract:

bstract:Objective
To investigate the inhibitory effect of constitutive  photomorphogenic homolog subunit 3 of COP9(COPS3) gene interferred by RNA on

the proliferation of lung cancer A549 cells, and to clarify its mechanism. Methods   The tumor experimental models of nude mice

were established in lung cancer  cell line A549.40 nude mice were divided into experiment group (n=20,implanted with A549 cells

infected by si-COPS3) and  control group(n=20,implanted with A549 cells infected by si-CTRL.The tumor size,weight, and tumor

formation rate of the nude mice  were detected.The expression levels of the cell cycle related proteins,such as

P21,CDK2,CDK4,cyclinB1, and cyclinD1 were detected by Western blotting method after down-regulation of expresion of COPS3 gene in

A549 cells. Results The tumor volume and weight(P<0.05),and  the tumor formation rate  of nude mice in experiment group were

significantly lower  than those in  control group.Compared with control group,the expression level of P21 protein was up-regulated

(P<0.05),and the expression levels  of CDK4 and cyclinB1 protein were  down-regulated(P<0.05),while the expression levels of

CDK2 and cyclinD1 protein had no significant changes(P=0.384,P=0.605).Conclusion The proliferation  of lung cancer cells can be

suppressed significantly by inhibiting the  expression of COPS3 gene.This effect may be mediated by up-regulating the expression of

P21 and down-regulating the expressions of cyclinB1 and CDK4.

Key words: constitutive , photomorphogenic homolog subunit 3 of COP9;RNA interference;lung neoplasms;lentivirus vector

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