甲状腺癌甲基化差异片断的筛选

Isolation of methylative difference DNA fragments in thyroid cancer

目的筛选甲状腺癌与甲状腺癌旁组织的甲基化差异片段。方法采用甲基敏感性代表性差异分析（MS-RDA）方法,用甲基化敏感性内切酶消化甲状腺癌及癌旁组织基因组DNA,连上接头制备扩增子后,进行两轮消减杂交,筛选甲状腺癌与癌旁组织的甲基化差异片段。结果经过两轮消减杂交后,筛选出3个低甲基化差异片段,2个高甲基化差异片断。筛选出的低甲基化差异片段C913位于BC029276基因上,该基因编码一种rRNA结合蛋白,可能与肿瘤有关。另外2个低甲基化差异片段C915与C917尚未见相关结构和功能的报道。两个高甲基化差异片断与已知基因同源性在20%-35%之间。5个片断均为新的甲基化差异片断。结论在甲状腺癌组织中存在异常甲基化,筛选出的甲基化差异片断有助于进一步了解异常甲基化在甲状腺癌发生发展中的作用。

Objective To investigate the aberrantly methylated status in human thyroid cancer. Methods Methylation - sensitive - representational difference analysis （MS - RDA） technique was employed to the primary thyroid cancer and adjacent noncancerous tissue. Results Following two sueeessive competitive hybridization, 3 hypo - and 2 hypermethylated DNA fragments was identified. One of these 3 hypomethylated fragments, C913, is located in BC029276 gene which encode a rRNA binding protein and may be associated with neoplasm. The strueture and function of the other two, C915 and C917, is not clear at present. The 2 hypermethylated fragments, （2919, C922, showed low homology with any known gene in GenBank. Conclusion Aberrant methylation may involve in development of thyroid cancer.