结晶型硫化镍及反式-BPDE恶性转化16HBE细胞hMSH2基因甲基化的研究  

Aberrant Methylation of the hMSH2Gene in the Nickel Sulfide and Anti-BPDE Transformed 16HBE Cells

作　　者：刘莉莉[1] 陈家[1] 安社娟[1] 吴中亮[1] 

机构地区：[1]广州医学院化学致癌研究所,广东广州510182

出　　处：《癌变．畸变．突变》2005年第3期129-132,共4页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：国家自然科学基金资助项目(No.39970630)

摘　　要：背景与目的:对结晶型硫化镍(Nickelsulfide,NiS)及反式二氢二醇环氧苯并芘(anti_7,8,_dihydrodiol_9,10_epoxidebenzo[a]pyrene,BPDE)恶性转化及成瘤的人支气管上皮细胞(Humanbronchialepithelial,16HBE)hMSH2基因启动子甲基化状况及其mRNA表达进行研究,探讨镍及反式_BPDE的表遗传致癌机制。材料与方法:采用甲基化特异性PCR(Methylation_specificPCR,MSP)法和RT_PCR法检测结晶型NiS及反式_BPDE恶性转化及成瘤的16HBE细胞hMSH2基因启动子甲基化状况及其mRNA表达,与非转化的16HBE细胞进行比较;并用去甲基化因子5_Azac(5_Aza_2′_deoxycytidine)处理有异常甲基化的细胞。结果:发现结晶型NiS及反式_BPDE恶性转化及成瘤的16HBE细胞hMSH2基因启动子区存在CpG岛的高甲基化;与非转化16HBE细胞比较,转化及成瘤的16HBE细胞hMSH2基因mRNA表达下降;有异常甲基化的细胞经去甲基化处理后甲基化消失。结论:结晶型NiS及反式_BPDE恶性转化及成瘤的16HBE细胞hMSH2基因启动子区CpG岛的高甲基化使其mRNA表达下降,并可能导致hMSH2基因表达抑制,这可能是结晶型NiS及反式_BPDE诱导16HBE细胞转化和成瘤的一种表遗传致癌机制。甲基化的可逆性对今后研究其表型逆转以及药物治疗提供了重要依据。BACKGROUND&AIM: To study the aberrant methylation of the hMSH2gene promoter and its mRNA expression in the nickel sulfide(NiS)and anti_7,8,_dihydrodiol_9,10_epoxide benzo[a] pyrene(anti_BPDE)transformed16HBE cells and to explore the possible epigenetic mechanism for NiS and anti_BPDE carcinogenesis. MATERIAL AND METHODS: DNA methylation patterns in the hMSH2gene promoter were determined by methylation_specific PCR(MSP)assay and mRNA expression was analysed by RT_PCR assay.The results were compared with the non_transformed16HBE cells which and the aberrant methylation cells were treated with demethylating agent5_Aza_2′_deoxycytidine. RESULTS: The hypermethylation in CpG island of the hMSH2gene promoter was indentified in the NiS and anti_BPDE transformed16HBE cells;comparing with non_transformed cells,hMSH2gene mRNA expression levels of the NiS and anti_BPDE transformed16HBE cells were reduced;treatment of the hMSH2with5_Azac decreased the methylation. CONCLUSION: Hypermethylation in CpG island of the hMSH2gene promoter is known to result in mRNA expression reducing and gene silencing probably,it may represent a possible epigenetic mechanism for NiS and anti_BPDE induced cells transformation and carcinogenesis.Reversible methylation offered an important evidence for phenotype inversion and drug treatment.

关 键 词：HMSH2 基因甲基化 硫化镍 反式-二氢二醇环氧苯并茈 

分 类 号：R994.3[医药卫生—毒理学]