结直肠癌和管状腺瘤中Runx3基因启动子甲基化状态的分析  

Methylation status of Runx3 gene promoter in colorectal cancer and tubular adenoma

作　　者：石明伟[1] 张侠[1] 许春伟[2] 张玉萍[2] 

机构地区：[1]安徽医科大学北京军区总医院临床学院肿瘤科,北京100700 [2]安徽医科大学北京军区总医院临床学院病理科,北京100700

出　　处：《临床与实验病理学杂志》2013年第12期1283-1287,共5页Chinese Journal of Clinical and Experimental Pathology

摘　　要：目的探讨结直肠癌及管状腺瘤中Runx3基因甲基化状态和Runx3蛋白表达,确定Runx3基因在"腺瘤→腺癌"癌变通路中的作用、临床意义及Runx3基因在不同年龄层段的甲基化程度。方法应用TaqMan探针qPCR(MethyLight)法分别检测40例管状腺瘤组织、50例结直肠癌组织和20例正常组织中Runx3基因CpG岛甲基化状态,并通过测序法验证扩增序列,同时应用免疫组化法分别检测上述标本中Runx3蛋白表达。结果结直肠癌组、管状腺瘤组、正常组中Runx3基因启动子甲基化率分别为84%、45%、10%,差异有统计学意义(P<0.05);结直肠癌组、管状腺瘤组、正常组中Runx3蛋白阳性率分别为18%、45%、80%,差异有统计学意义(P<0.05);Runx3基因甲基化与蛋白表达在正常组、管状腺瘤组及结直肠癌组中均呈负相关(相关系数r值分别为-0.667、-0.616、-0.790,P均<0.05);管状腺瘤组和结直肠癌组中Runx3基因启动子甲基化频率与患者年龄呈正相关(相关系数r值分别为0.376、0.304,P均<0.05)。结论 Runx3基因甲基化可能诱导其蛋白表达下调,在结直肠"腺瘤→腺癌"的癌变通路中起重要作用,同时Runx3基因甲基化可能与患者年龄相关,随年龄增长,甲基化程度增高。Purpose To investigate the expression and methylation status of the Runx3 gene in the colorectal carcinoma and tubular adenoma, to verify the function and clinical significance of the Runx3 gene in the pathway of adenoma to adenocarcinoma, and to ana-lyze the levels of methylation of Runx3 in different age groups. Methods TaqMan probe qPCR （MethyLight） was used to detect the methylation status of CpG island of Runx3 gene, and sequencing strategy to validate sequencing of amplified, and immunohistochemical staining to test the protein expression of Runx3 gene in 40 cases of tubular adenoma, 50 cases of colorectal cancer, and 20 cases of nor-mal colorectal mucosa tissues. Results The methylation rates of Runx3 gene promoter were 84% , 45% , and 10% （P 〈 0. 05 ） , and the protein expression rates of Runx3 gene were 18%, 45%, and 80% （ P 〈 0. 05 ） in the groups of colorectal carcinoma, tubular ade- noma and normal tissues, respectively. A significant inverse correlation was found between the methylation rate of Runx3 gene and the protein expression rate of Runx3 gene in the three groups （ Pearson＇ s correlation coefficients ： r = - 0. 667, r = - 0. 616, r = - 0. 790, respectively, all P 〈 0. 05 ）. A significantly positive correlation was found between the methylation rate of Runx3 gene and age in the groups of tubular adenoma and colorectal carcinoma （ Pearson＇ s correlation coefficients ： r = 0. 376, r = 0.304, respectively, both P 〈 0. 05 ）. Conclusion The methylation of Runx3 gene may result in decreased protein expression, which plays an important role in the pathway of adenoma to adenocarcinoma. And the methylation of Runx3 gene may correlate with ages ： along with aging, the methylation level increases.

关 键 词：DNA甲基化 RUNX3基因 qPCR DNA探针 

分 类 号：R735.3[医药卫生—肿瘤]