CHD5基因在肺癌细胞系中的表达水平及其CpG岛的甲基化分析.doc

文档介绍：
CHD5 基因在肺癌细胞系中的表达水平及其
CpG 岛的甲基化分析#
郑文岭,马文丽**
5
10
(南方医科大学基因工程研究所)
摘要: 目的: 明确肺癌细胞中人肿瘤相关基因 CHD5 基因在甲基化转移酶抑制剂
5-Aza-2'-deoxycytidine(AZA)作用前后的表达及其甲基化情况,探讨 CHD5 基因的肿瘤抑制
机制。方法:本文采用荧光定量 PCR 方法检测了 H1299, A549, H1650, H358, HCC827,
H460,H661 肺癌细胞系中 CHD5 的表达水平,,并利用 Western-blot 技术检测了上述样本的
蛋白表达水平。用 Bisulfite Sequencing PCR 方法检测 CHD5 的 CpG 岛区的甲基化水平。结
果:7 例肺癌细胞系与正常肺组织细胞相比,所有肺癌细胞系(7/7)CHD5 无论是在 mRNA
表达水平还是在蛋白表达水平均有明显下降。而在检测肺癌细胞系(A549, H1299, and H460)
启动子区域存在大量的甲基化位点。结论:CHD5 基因在肺癌细胞中表达是受其甲基化调控
的。
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关键词:细胞生物学;CHD5;甲基化;肺癌细胞
中图分类号:Q2;Q7
Analysis of Methylation of the CHD5 Gene in Human Lung
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25
30
Cancer cell
zheng wenling, ma wenli
(Southern medical university ,Guangzhou ,510515)
Abstract: Objective: To discover the tumor-suppressive mechanisms in human colon carcinoma.
Methods: CHD5 mRNA level of seven cases lung cancer cell lines (H1299, A549, H1650, H358,
HCC827, of H460, H661) were determined by quantitative RT-PCR. Additionally, these samples
were detect the protein expression level by Western-blot technique. The genome-wide
hypomethylation and the abnormal methylation of CpG island were mon phenomenon in
human status of CHD5 genes was determined by bisulfite sequencing PCR
( BSP).Results:
All cell lines examined exhibited significantly lower levels of CHD5 gene expression relative to
normal lung tissues. The lung cancer cell li