Disabled-2基因去甲基化对肺癌细胞系Wnt通路及增殖能力的影响  

The effect of Wnt pathway and proliferation of lung cancer cells by demethylated Disabled-2 gene

作　　者：杨连赫[1] 马爽[2] 张婉琳 沈帅[1] 于承仟 王雪[1] Yang Lianhe Ma Shuang Zhang Wanlin Shen Shuai Yu Chengqian Wang Xue(Department of Pathology, the First Affiliated Hospital and College of Basic Medical Sciences of China Medical University ,Liaoning Shen- yang 110001, China Department of Neurology, Shengfing Hospital of China Medical University, Liaoning Shenyang 110004, China.)

机构地区：[1]中国医科大学附属第一医院病理科、中国医科大学基础医学院病理学教研室,辽宁沈阳110001 [2]中国医科大学附属盛京医院神经内科,辽宁沈阳110004

出　　处：《现代肿瘤医学》2016年第23期3696-3699,共4页Journal of Modern Oncology

基　　金：国家自然科学基金项目(编号:81301930);辽宁省教育厅科学研究一般项目(编号:L2015595);大学生创新训练项目(甲类)(编号:201610159000036)

摘　　要：目的:探讨Dab2启动子区的甲基化状态及其对Dab2表达的影响,对Wnt通路和肺癌细胞增殖能力的影响。方法:BSP检测去甲基化试剂处理前后BE1细胞启动子区Dab2甲基化状态;Real-time PCR检测去甲基化试剂处理前后Dab2 mRNA水平;Western blot检测去甲基化试剂处理前后Dab2变化和Wnt通路的变化;MTT法检测BE1细胞经去甲基化试剂处理后增殖能力的变化。结果:BE1细胞Dab2基因启动子区处于高甲基化状态,经去甲基化试剂处理后Dab2启动子区发生明显去甲基化(37%vs 10.3%,P<0.01),Dab2mRNA在去甲基化试剂处理后明显上调,Western blot检测显示BE1细胞经去甲基化试剂处理后Dab2蛋白表达明显上调,而Wnt通路关键分子β-catenin及其下游Cyclin D1表达下调,BE1细胞经去甲基化试剂处理后增殖能力明显下调。结论:Dab2基因启动子区高甲基化抑制肺癌细胞中Dab2转录水平和表达,Dab2基因去甲基化抑制Wnt通路,进而抑制肺癌细胞的增殖能力。Objective： This paper is aim to study the methylation status of Dab2 promoter region and its relationship with Dab2 expression,as well as its affection to Wnt pathway and proliferation in lung cancer cell line. Methods：BSP was used to detect the methylation status of Dab2 promoter region in BE1 cell with or without demethylating agent. Real- time PCR was used to test the Dab2 mRNA level. Western blot was used to detect the change of Dab2 and Wnt pathway. MTT was used to detect the change of proliferation of BE1 cell line after demethylating agent. Results：Abnormal hypermethylation could be found in Dab2 promoter region,it could be demethylated after demethylating agent（ 37% vs 10. 3%,P〈0. 01）. Real- time PCR showed that Dab2 mRNA was up- regulated after demethylating agent treatment. Western blot showed Dab2 up- regulation and down- regulation of β- catenin and Cyclin D1. MTT result indicated the decreasing proliferated ability after demethylating agent in BE1 cell line. Conclusion： Hypermethylated Dab2 promoter inhibits the Dab2 transcription and expression level in lung cancer cell,demethylated Dab2 might inhibit Wnt pathway,as well as the proliferation of lung cancer cell.

关 键 词：甲基化 肺癌 增殖 

分 类 号：R730.23[医药卫生—肿瘤]