DR4基因启动子甲基化对TRAIL诱导肺鳞癌细胞凋亡作用的影响  

Effect of DR4 promoter methylation on the TRAIL induced apoptosis in lung squamous carcinoma cell

作　　者：余宗阳[1] 赵忠全[1] 黄晓燕 潘燕 施燕鸿 吴晓龙 林祥武 陈思玉 陈颖[1] 王水良[1] 綦晓艳 王文武[1] Yu Zongyang;Zhao Zhongquan;Huang Xiaoyan;Pan Yan;Shi Yanhong;Wu Xiaolong;Lin Xiangwu;Chen Siyu;Chen Ying;Wang Shuiliang;Qi Xiaoyan;Wang Wenwu(Department of Oncology,Fuzhou General Hospital of PLA,Fujian Fuzhou 350025,China;Department of Oncoology,Central Hospital of Zibo,Shandong Zibo 255036,China.)

机构地区：[1]解放军福州总医院肿瘤科,福建福州350025 [2]淄博市中心医院肿瘤科,山东淄博255036

出　　处：《现代肿瘤医学》2018年第20期3205-3209,共5页Journal of Modern Oncology

基　　金：福建省自然科学基金项目(编号:2017J01322)

摘　　要：目的:探讨肺鳞癌中死亡受体4(DR4)不同甲基化状态是否会影响肿瘤坏死因子相关凋亡诱导配体(TRAIL)对肺鳞癌细胞的诱导凋亡作用。方法:采用甲基化特异性PCR(MSP)、RT-PCR和Western blot法检测5-氮-2'-脱氧胞苷(5-Aza-CdR)处理肺鳞癌细胞株(H226、SK-MES-1、H520)前后DR4基因启动子甲基化状态和基因表达情况;MTT法检测细胞增殖抑制率,流式细胞仪检测细胞凋亡率。结果:MSP、RT-PCR和Western blot结果表明H226、SK-MES-1细胞DR4基因呈甲基化状态,其mRNA及蛋白均低表达,H520细胞中DR4基因呈非甲基化状态,其mRNA及蛋白高表达;5-Aza-CdR干预后H226、SK-MES-1细胞DR4基因呈非甲基化状态,其mRNA及蛋白表达较前均显著上调,差异有统计学意义(P<0.05),H520仍呈非甲基化状态,其mRNA及蛋白表达量较前无明显差异。同时研究还证实TRAIL对H226、SK-MES-1细胞有不同程度的细胞增殖抑制率和凋亡率,但不敏感,5-Aza-CdR干预后,H226及SK-MES-1细胞对TRAIL的敏感性较前均显著增高(P<0.05)。结论:5-Aza-CdR可以逆转肺鳞癌中DR4基因启动子甲基化状态,进而增加TRAIL诱导肺鳞癌细胞凋亡的作用。5-Aza-CdR联合TRAIL可能是治疗肺鳞癌的一种新策略。Objective：To discuss the silence of DR4 is associated with gene methylation,the article discusses DR4 gene promoter methylation with the expression of DR4 in lung squamous carcinoma, and we further investigated on whether DR4 methylafion status would affect the effect of the tumor necrosis factor related apoptosis inducing ligand （TRAIL） induced apoptosis of lung squamous cancer cells. Methods： Methylation specific PCR（MSP）, liT- PCR and Western blot method were used to detect DR4 gene promoter methylation status and the expression of DR4 in 5-N-2 ＇-deoxy cytidine （ 5 -Aza-CdR） treated pulmonary squamous cancer cells （ H226, SK-MES-1, H520）. The MTT method was adopted to detect cell proliferation inhibition rate, and the flow cytometry to detect cell apoptosis rate. Results：MSP,RT-PCR and Western blot results showed that DR4 gene presented methylation status in H226, SK-MES-1 cell,with low mRNA and protein expression,while H520 cell showed opposite results. In 5-Aza-CdR treated H226, SK-MES-l cells, DR4 gene exhibit unmethylation status, and its mRNA and protein expression were significantly raised（ P 〈 0.05 ）. At the same time the study also confirmed that the TRAIL can induce cell proliferation inhibition and apoptosis in H226 ,SK-MES-1 cell in different degree,5-Aza-CdR can increase H226 and SK-MES-1 cell sensitivity to the TRAIL significantly（ P 〈 0.05 ）. Conclusion： Lung squamous carcinoma of DR4 high methylafion modification can result in DR4 silence or lower expression, while 5-Aza-CdR can reverse DR4 gene promoter methylation status, it can also increases the TRAIL induced pulmonary squamous cancer cells apoptosis, therefore 5-Aza-CdR joint TRAIL may be a new strategy for the treatment of lung squamous carcinoma.

关 键 词：肺鳞癌 5-AZA-CDR TRAIL DNA甲基化 DR4基因 

分 类 号：R734.2[医药卫生—肿瘤]