摘要
目的研究缺氧对人小细胞肺癌H446细胞基因组DNA甲基化水平的影响。方法体外培养人小细胞肺癌细胞H446,置于缺氧(3%O2)条件下培养,分别于0、12、24、48、72 h后提取细胞基因组DNA,用甲基化敏感性随机引物PCR(MS-AP-PCR)方法检测基因组DNA甲基化状态。结果在缺氧状态下,H446细胞经酶解后的扩增谱带明显增多,且荧光强度较强,以24 h后明显,表明H446细胞基因组DNA甲基化水平在缺氧24 h后即发生明显升高,主要表现为多位点的异常甲基化。结论缺氧可能是造成肿瘤细胞内甲基化失衡的重要原因之一。
Objective To investigate the influence of hypoxia on the methylation level of genomic DNA in human small cell lung cancer H446 cells.Methods H446 cells were cultured under the hypoxic condition(3%O2) for different time periods(0,12,24,48 and 72 h respectively).Those cultured under normal condition served as control.The genomic DNA methylation level was detected by methylation sensitivity random primers PCR(MA-AS-PCR) after DNA extraction.Results Compared with the normoxic control group,the quantity and fluorescence intensity of amplified products of hypoxic experimental group were much higher,especially after 24 h.This indicated that the genomic DNA methylation level in the H446 cells was significantly changed when were treated by hypoxia,and represented mainly as multifocal abnormal methylation.Conclusion Hypoxia might be one of the key causer inducing unbalanced methylation in cancer cells.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2011年第20期2145-2147,共3页
Journal of Third Military Medical University
基金
成都军区医学科研计划课题(MB09010)~~
关键词
缺氧
癌
小细胞
DNA甲基化
细胞系
肿瘤
hypoxia
carcinoma
small cell
DNA methylation
cell line
tumor