Disabled-1在人乳腺癌细胞中的表达及其对细胞周期的影响  

Expression of Disabled-1 in human breast cancer cells and its role in cell cycle

作　　者：曹让娟[1] 李凯[2] 邢琬莹[3] 王悦书[1] 于维[1] 吴广智[1] 崔树森[1] 李强[1] 

机构地区：[1]吉林大学中日联谊医院手外科,吉林长春130033 [2]吉林大学中日联谊医院麻醉科,吉林长春130033 [3]吉林大学中日联谊医院乳腺外科,吉林长春130033

出　　处：《吉林大学学报（医学版）》2016年第5期932-936,共5页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(81502516);吉林省科技厅科技发展计划项目资助课题(20160101077JC);吉林省科技厅医药产业发展引导资金项目资助课题(20150311069YY);吉林大学白求恩计划项目青年基金资助课题(2015429)

摘　　要：目的:探讨Disabled-1(Dab1)基因在乳腺上皮细胞及乳腺癌细胞中的表达情况,并阐明Dab1对乳腺癌细胞周期的影响。方法:体外培养永生化的乳腺上皮细胞MCF-10A及乳腺癌细胞MCF-7、BT-549和MDA-MB-231,实时荧光定量(Real-time PCR)法检测MCF-10A、MCF-7、BT-549和MDA-MB-231细胞中Dab1mRNA的表达水平,Western blotting法检测Dab1蛋白的表达水平。取对数生长期的BT-549细胞,分为对照组、转染pKH3组和转染pKH3-Dab1组,采用流式细胞术检测各组细胞周期的变化情况。结果:Real-time PCR法检测,与MCF-10A细胞(0.998±0.020)比较,乳腺癌细胞MCF-7(0.504±0.037)、BT-549(0.302±0.027)和MDA-MB-231(0.330±0.031)中Dab1mRNA相对表达水平明显下降(P<0.05)。Western blotting法检测,与MCF-10A细胞(0.227±0.021)比较,乳腺癌细胞MCF-7(0.134±0.014)、BT-549(0.076±0.01)和MDA-MB-231(0.074±0.005)中Dab1蛋白表达水平明显下调(P<0.05)。流式细胞术检测,与对照组和转染pKH3组比较,转染pHK3-Dab1组BT-549细胞出现细胞周期G1期阻滞。结论:Dab1在体外培养乳腺癌细胞系中表达下调,Dab1过表达可阻滞乳腺癌细胞G1细胞周期进展。Objective：To explore the expressions of Disabled-1 （Dab1 ）in human breast epithelial cells and breast cancer cells,and to clarify its role in cell cycle.Methods：Real-time PCR was used to analyze the Dab1 mRNA expressions in breast epithelial cells MCF-10A and breast cancer cells MCF-7,BT-549,and MDA-MB-231. The Dab1 protein expressions in those cells were tested by Western blotting method. The BT-549 cells at logarithmic growth phase were divided into control,pKH3,and pKH3-Dab1 groups;the cell cycle was investigated by flow cytometry.Results：The Real-time PCR results showed that the Dab1 mRNA expression levels in MCF-7＆amp;nbsp;cells （0.504 ± 0.037）,BT-549 cells （0.302 ± 0.027）,and MDA-MB-231 cells （0.330 ± 0.031 ）were reduced compared with MCF-10A cells （0.998±0.020）（P 〈0.05）.The Western blotting results showed that the Dab1 protein expression levels in breast cancer cells MCF-7 （0.134±0.014）,BT-549 （0.076±0.01）,and MDA-MB-231 （0.074±0.005）were reduced compared with MCF-10A cells （0.227±0.021）（P 〈0.05）.Compared with control group and pKH3 group,the cell cycle in pKH3-Dab1 group was inhibited at G1 phase detected by flow cytometry analysis. Conclusion：The expression of Dab1 is down-regulated in breast cancer cells,and the over-expression of Dab1 can inhibit the cell cycle at G1 phase.

关 键 词：Dab1基因 人乳腺癌细胞 细胞周期 G1期阻滞 

分 类 号：R730.2[医药卫生—肿瘤]