肝癌组织中ZHX2基因启动子甲基化及其与临床病理特征的关系

摘要

Objective Denaturing high performance liquid chromatography assay (DHPLC) was used to detect promoter methylation of ZHX2 gene in primary liver cancer, so as to further explore the relationship between promoter methylation of ZHX2 gene with clinical and pathological features of hepatocellular carcinoma Methods Methylated UNA in vitro and human placenta DNA were methylated and nonmethylated control groups respectively. By bisulfite-DHPLC technology, DHPLC method was established to analyze promoter methylation of ZHX2. Under the conditions of partially denatured temperature at 55 ℃, promoter methylation of ZHX2 gene was detected. And 24 cases of HCC and adjacent liver tissues and 6 normal liver tissue samples were detected. Results DHPLC analysis to promoter methylation ZHX2 was established. Using this method, no methylation was detected in normal liver tissue samples, 3 adjacent tissue ( 16.6% ) had methylation, 11 cases were detected methylation in 24 HCCs. Conclusions DHPLC method is first established to detect promoter methylation of ZHX2 gene in liver tissue, which gives the foundation for further research the relationship between ZHX2 gene and development of liver cancer.%目的 运用变性高效液相色谱(Denature High-Performance Liquid Chromatography,DHPLC)法检测原发性肝癌细胞中ZHX2基因启动子甲基化的情况,为进一步探讨其与原发性肝细胞癌(HCC)临床病理特征之间的关系奠定基础.方法 采用亚硫酸氢钠-DHPLC技术,以体外甲基化的DNA和人类胎盘DNA分别为甲基化和非甲基化对照,建立ZHX2基因启动子甲基化DHPLC分析方法.在55℃部分变性条件下,对ZHX2基因启动子重要区域甲基化水平进行检测,并对24例HCC癌组织及癌旁肝组织和6例正常肝组织标本进行检测.结果 建立了ZHX2基因启动子甲基化DHPLC分析方法,使用该方法检测结果显示正常肝组织未检测到甲基化,24例HCC癌组织中有11例存在甲基化(45.8％),而24例HCC癌旁组织中有3例(16.6％)存在甲基化.结论 本研究建立了ZHX2基因启动子甲基化DHPLC分析方法,并首次将其应用于肝癌组织标本的检测,为深入探讨ZHX2基因与HCC发生发展之间关系奠定了基础.