中国全科医学2012，Vol.15Issue(6)：644-648,5.
甲基化CpG岛扩增联合代表性差异分析方法在筛查人非小细胞肺癌组织中高甲基化修饰序列的应用
Identification of Hyper-Methylated Sequences in Lung Cancer by Methylated CpG Island Amplification Coupled with Representational Difference Analysis
摘要
Abstract
Objective To identify the promoter hypermethylation of CpG islands in human non - small - cell lung cancer ( NSCLC ) . Methods Methylated CpG island amplification ( MCA ) coupled with Representational Difference Analysis ( RDA ) were used to isolate methylated CpG islands in 92 NSCLC tumor tissues and adjacent normal tissues. RT - PCR and meth-ylation - sensitive PCR ( MSP ) were used to examine the correlation between CpG islands methylation and gene expression of IG-FBP - 4. Results Five sequences that corresponded to promoter CpG islands of IGFBP - 4, KLK10 , ADAM23 , GADD45 G and DU0X1 genes were isolated and confirmed by Slot Blot hybridization in NSCLC specimens. Weak expression of IGFBP - 4 in NSCLC cells was increased after the treatment with 5 - aza - 2' - deoxycytidine. The expression of IGFBP - 4 was significantly downregulated in 41 ( 44. 6% ) tumor tissues compared with corresponding normal tissues. IGFBP -4 hypermethylation was detected in 47 ( 51. 1 % ) tumor tissues, significantly higher than normal tissues ( 16. 3% , 15/92, P <0. 001 ) . IGFBP -4 expression and its promoter hypermethylation were negatively correlated (r= -0.396, P<0.001 ) .Moreover, IGFBP-4 hypermethylation was associated with advanced stage ( P = 0. 013 ) and lymph metastasis ( P = 0. 022 ) . Conclusion Five hypermethylat-ed CpG islands were isolated by MCA/RDA method in NSCLC specimens and epigenetic silencing by hypermethylation of the IGFBP - 4 CpG - rich promoter region is likely to be involved in the progression of NSCLC.关键词
癌,非小细胞肺/甲基化CpG岛扩增/代表性差异分析/胰岛素样生长因子结合蛋白质4Key words
Carcinoma/ non - small - cell lung/ Methylated CpG island amplification/ Representational difference analysis/ Insulin - like growth factor binding protein 4分类
医药卫生引用本文复制引用
李勇,郭榕,宁金鹰..甲基化CpG岛扩增联合代表性差异分析方法在筛查人非小细胞肺癌组织中高甲基化修饰序列的应用[J].中国全科医学,2012,15(6):644-648,5.基金项目
国家自然科学基金(30871366) （30871366）
