摘要
背景与目的TSLC1在多种肿瘤中表达下调或失活,其表达下调与DNA高甲基化有很大关系。本研究旨在探索TSLC1在肺癌细胞中的表达缺失与其启动子区DNA甲基化的关系。方法采用RT-PCR和Real-time PCR方法检测TSLC1在正常肺组织和3种肺癌细胞系(A549、NCI-H446和Calu-3)中的表达谱;运用亚硫酸氢盐修饰后测序(bisulfite sequencing)方法检测上述正常肺组织和肺癌细胞中TSLC1启动子区的甲基化状态;应用甲基化转移酶抑制剂5-氮-2-脱氧胞苷(5-Aza-dC)处理上述细胞后,采用Real-time PCR方法检测处理前后TSLC1的表达变化。结果TSLC1在正常肺组织和A549细胞中表达,其启动子区DNA未发生甲基化;而在NCI-H446和Calu-3细胞中表达缺失,其启动子区DNA发生高甲基化,并且5-Aza-dC处理NCI-H446和Calu-3细胞后可促进TSLC1的表达。结论TSLC1在肺癌细胞中的表达缺失是由其启动子区的DNA高甲基化引起。
Background and objective The expression of TSLC1 is downregulated or abrogated in many kinds of tumors, and its downregulation is highly associated with DNA hypermethlyation. The aim of this study is to explore the rela-tionship between TSLC1 silencing and DNA methylation of its promoter region in lung cancer cells. Methods We detected the expression pattern of TSLC1 in human normal lung tissue and three lung cancer cell lines （A549, NCI-H446 and Calu-3） by semi-quantitative RT-PCR and Real-time PCR. Then we detected the status of DNA methylation in TSLC1 promoter re-gion with bisulfite sequencing in above normal lung tissue and lung cancer cell lines. After treatment of above cell lines with the inhibitor of DNA methyltransferase 5-Aza-2-deoxycytidine （5-Aza-dC）, we detected the expression change of TSLC1 by Real-time PCR before and after the treatment of 5-Aza-dC. Results There was no methylation in TSLC1 promoter region in normal lung tissue and A549 cell line in which TSLC1 expressed; while there was DNA hypermethylation in TSLC1 promoter region in NCI-H446 and Calu-3 cell lines in which TSLC1 was abrogated, also the expression of TSLC1 in NCI-H446 and Calu-3 cell lines could be restored after treatment of 5-Aza-dC. Conclusion The silencing of TSLC1 in lung cancer cells is due to the hypermethylation of its promoter region.
出处
《中国肺癌杂志》
CAS
2010年第5期464-469,共6页
Chinese Journal of Lung Cancer
