CAC1对胃癌细胞系AGS细胞周期的调控作用  

Exploration on the role of CAC1 in the regulation of cell cycle of AGS gastric cancer cell line

作　　者：郑琪[1] 张茜[1] 张彧[1] 廖子君[1] 姚煜[2] 南克俊[2] 

机构地区：[1]西安交通大学医学院附属陕西省肿瘤医院内一科,西安710061 [2]西安交通大学第一附属医院肿瘤内科

出　　处：《山西医科大学学报》2016年第1期35-40,共6页Journal of Shanxi Medical University

摘　　要：目的研究新基因CAC1对胃癌细胞系AGS的细胞周期和细胞周期相关基因表达的调控作用。方法将AGS细胞分为空白对照组、阴性对照组和干扰组,分别给予细胞培养液、阴性对照siRNA和有效的CAC1-siRNA处理,用流式细胞仪检测细胞周期的变化,实时定量PCR和Western blot法检测细胞周期相关基因(p53、cyclin A、cyclin E及CDK2)在mRNA和蛋白水平的表达变化。结果 CAC1沉默后,流式细胞仪检测发现,RNA干扰组的AGS细胞G1期细胞比例[(73.23±3.04)%]较对照组[(45.33±0.82)%]明显增加(P<0.05),S期细胞比例[(5.40±5.83)%]较对照组[(41.07±1.07)%]显著减少(P<0.05)。CAC1沉默后,与对照组相比,干扰组无论是cyclin E的mRNA表达水平(1.000±0.000 vs 0.294±0.011,P<0.05),还是cyclin A的mRNA表达水平(1.000±0.000 vs 0.886±0.039,P<0.05),均出现了明显的下降,p53 mRNA表达水平显著上升(1.000±0.000 vs 2.233±0.122,P<0.05),CDK2mRNA表达水平无明显变化(1.000±0.000 vs 0.952±0.007,P>0.05)。类似地,CAC1沉默后,与对照组相比,干扰组的cyclin E蛋白表达水平(0.667±0.236 vs 0.165±0.046,P<0.05)和cyclin A蛋白表达水平(0.607±0.284 vs 0.208±0.029,P<0.05)均出现了明显的下降,P53蛋白表达水平出现了显著的上升(0.326±0.054 vs 0.656±0.106,P<0.05);而CDK2蛋白表达水平无明显变化(0.864±0.175 vs 0.717±0.100,P>0.05)。结论 CAC1可促进胃癌AGS细胞的G1/S期转换,下调p53的表达,上调cyclin E和cyclin A的表达。Objective To investigate the role of novel gene CAC1 in regulating cell cycle and cell cycle-related genes expression of gastric cancer cell line AGS. Methods AGS cells were divided into blank control group,negative control group and RNA interference group,which were treated with cell culture fluid,negative control siRNA and effective CAC1-siRNA,respectively. The cell cycle changes were analyzed by flow cytometry,and the expression of cell cycle-related genes（ p53,cyclin A,cyclin E and CDK2） at mRNA and protein levels were detected by real-time PCR and Western blot,respectively. Results After CAC1 expression was effectively silenced by RNA interference in AGS cells,the cell proportion in G1phase[（ 73. 23 ± 3. 04） %] increased remarkably compared with control group[（ 45. 33 ± 0. 82） %,P〈0. 05） ],and the cell proportion in S phase decreased significantly[（ 5. 40 ± 5. 83） % vs（ 41. 07± 1. 07） %,P〈0. 05]. In addition,both cyclin E mRNA expression（ 0. 294 ± 0. 011 vs 1. 000 ± 0. 000,P〈0. 05） and cyclin A mRNA expression（ 0. 886 ± 0. 039 vs 1. 000 ± 0. 000,P〈0. 05） were significantly down-regulated after CAC1 silencing,p53 mRNA expression increased significantly（ 2. 233 ± 0. 122 vs 1. 000 ± 0. 000,P〈0. 05）,and CDK2 mRNA expression had no significant change（ 0. 952 ± 0. 007 vs 1. 000 ± 0. 000,P〈0. 05）. Likewise,both cyclin E protein expression（ 0. 165 ± 0. 046 vs 0. 667 ± 0. 236,P〈0. 05） and cyclin A protein expression（ 0. 208 ± 0. 029 vs 0. 607 ± 0. 284,P〈0. 05） were dramatically down-regulated after CAC1 silencing,P53 protein expression increased（ 0. 656 ± 0. 106 vs 0. 326 ± 0. 054,P〈0. 05）,and CDK2 protein expression had no significant change（ 0. 717 ± 0. 100 vs 0. 864 ± 0. 175,P〈0. 05）. Conclusion CAC1 can promote G1/ S transition,down-regulate P53 expression and up-regulate cyclin E and cyclin A expression in AGS gastric cancer cell line.

关 键 词：胃癌 AGS细胞 CAC1 细胞周期 

分 类 号：R735.2[医药卫生—肿瘤]