GADD45α在膀胱癌组织和细胞中的表达及作用机制  

The expression and mechanism of GADD45αin bladder cancer tissues and cells

作　　者：袁刚军 韩娜 鲜鹏 袁方 李俊 张海燕[2] 唐显力 戴君勇 宋彦平 李元 刘南 YUAN Gangjun;HAN Na;XIAN Peng;YUAN Fang;LI Jun;ZHANG Haiyan;TANG Xianli;DAI Junyong;SONG Yanping;LI Yuan;LIU Nan(Department of Urology,Chongqing University Cancer Hospital,Chongqing 400030,China;Health Examination and Oncology Screening Center Affiliated to Chongqing University,Chongqing 400030,China)

机构地区：[1]重庆大学附属肿瘤医院泌尿外科,重庆400030 [2]重庆大学附属健康体检与肿瘤筛查中心,重庆400030

出　　处：《现代肿瘤医学》2022年第2期206-210,共5页Journal of Modern Oncology

基　　金：重庆市自然科学基金面上项目(编号:cstc2019jcyj-msxmX0420)。

摘　　要：目的:检测GADD45α基因在膀胱癌中的表达情况及作用机制。方法:分别使用免疫组化、荧光定量PCR和Western blot实验检测GADD45α基因在膀胱癌组织中的表达情况;用慢病毒包装质粒上调膀胱癌细胞中GADD45α的表达后,流式细胞仪分析膀胱癌细胞周期比例分布情况和用Western blot实验检测细胞周期蛋白激酶水平变化情况。结果:膀胱癌组织中GADD45α的表达水平较癌旁正常组织的表达降低;上调膀胱癌细胞中GADD45α的表达后,与control细胞相比,高表达GADD45α细胞的克隆大小和数目皆明显下降(86±12.3 vs 134±13.5,P<0.05);升高GADD45α表达后,膀胱癌细胞G_(2)期比例升高至56.54%,而G_(1)期比例则由82.87%下降至40.8%,S期比例由17.13%下降至2.66%,膀胱癌细胞周期被阻滞在G_(2)/M期,伴随细胞周期蛋白激酶cdc2/cyclinB1的表达下降。结论:GADD45α在膀胱癌中表达下降,GADD45α通过抑制cdc2/cyclinB1的表达活性以阻滞膀胱癌细胞周期在G_(2)/M期,抑制膀胱癌细胞的增殖。Objective:To study the expression and underlying mechanism of GADD45αin bladder cancer.Methods:Using IHC,quantitative PCR,and Western blot analyses to detect the expression of GADD45αin bladder cancer tissues and cells.After upregulating the expression of GADD45αin bladder cancer cells with lentiviral packaging plasmids,flow cytometry was used to analyze the distribution of bladder cancer cell cycle proportions and Western blot experiments were used to detect changes in cyclin kinase levels.Results:GADD45αwas downregulated both in bladder cancer tissues and cells.After upregulation the expression of GADD45αin bladder cancer cells,compared with control cells,the clone size and number of high-expressing GADD45αcells were significantly reduced(86±12.3 vs 134±13.5,P<0.05).After increasing GADD45αexpression,the proportion of bladder cancer cells in G_(2) phase increased to 56.54%.The proportion of G_(1) phase dropped from 82.87%to 40.8%,and the proportion of S phase dropped from 17.13%to 2.66%.Cell cycles were arrested in G_(2)/M phase,accompanying with the dramatic decrease of cdc2/cyclinB1 activities.Conclusion:GADD45αwas downregulated in bladder cancer,and GADD45αinhibited the activities of cdc2/cyclinB1 to arrest bladder cancer cell cycle in G_(2)/M phase.

关 键 词：GADD45α 膀胱癌 P53 细胞周期 

分 类 号：R737.14[医药卫生—肿瘤]