靶向Cyclin D1基因RNA干扰对肝癌细胞增殖和Mdm2等基因表达影响的研究  

Effects of RNA interference of targeting Cyclin D1 gene on hepatoma cell proliferation and Mdm2 gene expression

作　　者：张弘英[1] 胡树根 胡利琳[3] 丁浩[1] ZHANG Hongying;HU Shugen;HU Lilin;DING Hao(Department of Gastroenterology,Second Affiliated Hospital of Nanchang University,Nanchang,Jiangxi 330006,China;Xinjian District People＇s Hospital Nanchang,Jiangxi 330100,China;Medical School of Nanchang University,Nanchang,Jiangxi 330006,China)

机构地区：[1]南昌大学第二附属医院消化内科,南昌330006 [2]江西省南昌市新建区人民医院,330100 [3]南昌大学医学院,南昌330006

出　　处：《重庆医学》2018年第15期1981-1985,共5页Chongqing medicine

基　　金：国家自然科学基金资助项目(81300348);江西省自然科学基金资助项目(20151BAB205100)

摘　　要：目的探讨Cyclin D1表达下调对Mdm2等基因表达及人肝癌细胞增殖的影响。方法用脂质体将Cyclin D1-siRNA转染人肝癌细胞株Hep3B细胞。实验分为空白对照组、阴性对照siRNA组、Cyclin D1-siRNA组。采用RT-PCR和Western blot检测Cyclin D1、Mdm2、Mdm4、P53和P21表达,流式细胞仪检测细胞周期,MTT检测细胞活力,TUNEL检测细胞凋亡。结果与空白对照组和阴性对照siRNA组相比,Cyclin D1-siRNA组P53和P21表达上调(P<0.05),Cyclin D1、Mdm2和Mdm4表达下调(P<0.01);3组细胞周期G1、S及G2期均无明显差异(P>0.05);Cyclin D1-siRNA组较其他两组细胞活力明显减弱(P<0.01),细胞凋亡明显增强(P<0.01)。结论 Cyclin D1表达下调能抑制Mdm2和Mdm4的表达,并能上调P53和P21的表达;Cyclin D1表达下调能抑制肝癌细胞的增殖并促进其凋亡。Objective To investigate the effects of down-regulating Cyclin D1 expression on Mdm2 gene expression and proliferation of human hepatoma cells. Methods Cyclin D1-siRNA was transfected into the human hepatoma cell line Hep3B s with liposome.The experiment was divided into the blank control group,negative control siRNA （NC-siRNA） group and Cyclin D1-siRNA group.RT - PCR and Western blot were used to detect the expressions of Cyclin D1,Mdm2,Mdm4,P53 and P21.The cell cycle was measured by flow cytometer;the cellular activity was tested with MTT;the cell apoptosis was examined by TUNEL. Results Compared with the blank control group and NC-siRNA group,the expressions of P53 and P21 in the Cyclin D1-siRNA group were increased （ P 〈0.05）,while the expressions of Cyclin D1,Mdm2 and Mdm4 were decreased （ P 〈0.01）;there were no significant differences in the G 1,S and G 2 phases among 3 groups（ P 〉 0.05 ）;the cell vitality in the in the Cyclin D1-siRNA group was significantly weakened compared with the other two groups（ P 〈0.01）,while the cell apoptosis was significantly enhanced（ P 〈0.01）. Conclusion The down-regulation of Cyclin D1 gene expression could inhibit the expressions of Mdm2 and Mdm4,and up-regulates the expressions of P53 and P21.Down-regulating Cyclin D1 expression can inhibit the proliferation of liver cancer cells and promotes their apoptosis.

关 键 词：细胞周期蛋白D1 RNA干扰 HEP3B细胞 细胞增殖 MDM2 

分 类 号：R363[医药卫生—病理学]