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IRX1在胰腺癌中的表达及其启动子区甲基化状态
引用本文:卫巍,徐凌,王锋,何珊珊,杨丽娟,郭传勇,王兴鹏. IRX1在胰腺癌中的表达及其启动子区甲基化状态[J]. 中华胰腺病杂志, 2011, 11(5). DOI: 10.3760/cma.j.issn.1674-1935.2011.05.002
作者姓名:卫巍  徐凌  王锋  何珊珊  杨丽娟  郭传勇  王兴鹏
作者单位:同济大学附属第十人民医院肝胆胰内科,上海,200072
基金项目:国家自然科学基金,上海市科委基金
摘    要:目的 检测胰腺癌的易洛魁族同源盒基因(IRX1)的表达及其启动子区的甲基化状态,探讨两者间的相关性.方法 采用实时PCR法检测12例胰腺癌组织及6株胰腺癌细胞株的IRX1 mRNA 表达.基因序列分析IRX1基因启动子区结构.应用甲基化抑制剂5-氮杂-2'-脱氧胞苷(5-Aza-dC)处理胰腺癌细胞,采用甲基化特异性PCR(MSP)、非甲基化特异性PCR (USP)及实时PCR检测处理前后IRX1启动子甲基化状态和IRX1 mRNA表达.结果 胰腺癌组织IRX1 mRNA的表达量为0.31±0.11,显著低于癌旁正常胰腺组织的1.05±0.32(P <0.01).胰腺癌细胞AsPCl、BxPC3、Capan-2、PANC1、PaTu8988和SW1990的IRX1 mRNA表达量分别为0.36±0.08、0.34±0.16、0.37±0.11、0.25±0.06、0.31±0.04、0.36±0.02,均显著低于人肾上皮293细胞的1.03±0.28(P<0.05或<0.01).IRX1基因启动子区富含CpG岛.各胰腺癌细胞株IRX1基因启动子CpG岛对应位点均有甲基化,经5-Aza-dC处理后甲基化状态得以逆转,IRX mRNA的表达也得以恢复.结论 胰腺癌的IRX1 mRNA表达下降,与其IRX1基因启动子区CpG岛高甲基化状态相关.

关 键 词:胰腺肿瘤  启动子  甲基化  IRX1

Expression and methylation of Iroquois homeobox protein 1 in pancreatic cancer
WEI Wei,XU Ling,WANG Feng,HE Shan-shan,YANG Li-juan,GUO Chuan-yong,WANG Xing-peng. Expression and methylation of Iroquois homeobox protein 1 in pancreatic cancer[J]. CHINESE JOURNAL OF PANCREATOLOGY, 2011, 11(5). DOI: 10.3760/cma.j.issn.1674-1935.2011.05.002
Authors:WEI Wei  XU Ling  WANG Feng  HE Shan-shan  YANG Li-juan  GUO Chuan-yong  WANG Xing-peng
Abstract:Objective To investigate the expression of Iroquois homeobox protein 1 ( IRX1 ) gene and the hypermethylation status of its promoter in pancreatic cancer,and their relationship.Methods Real-time PCR was used to quantitatively detect IRX1 gene expression level of 12 sets of resected pancreatic cancer tissue and 6 sets of pancreatic cancer cell lines; gene sequences analysis was used to detect the structure of IRX1 promoter; DNA methylation inhibitor 5-Aza-2′-deoxycytidine (5-Aza-dC) was used in pancreatic cancer cell lines,and then the methylation of IRX 1 was measured by methylation-specific PCR (MSP) and unmethylation sequence-PCR (USP) methods.Results Expression of IRX1 mRNA in pancreatic cancer tissue was 0.31 ± 0.11,which were significantly lower than that in normal pancreatic tissue ( 1.05 ±0.32,P <0.01 ).IRX1 mRNA expression of AsPCl,BxPC3,Capan 2,PANCl,PaTu8988 and SW1990 were 0.36 ± 0.08,0.34 ±0.16,0.37 ±0.11,0.25 ±0.06,0.31 ±0.04,0.36 ±0.02,which were significantly lower than that in human kidney epithelial 293 cells ( 1.03 ± 0.28,P < 0.05 or < 0.01 ).Analysis of IRX1 gene sequence showed abundant CpG islands in promoter.Hypermethylation of IRX1 promoter site was found in all pancreatic cancer cell lines.However,its methylation status could be reversed by 5-Aza-dC,and the IRX1 expression was also restored.Conclusions IRX1 mRNA expression is down-regulated in pancreatic cancer,and it is related with promoter CpG islands hypermethylation.
Keywords:Pancreatic neoplasms  Promoter  Methylation  IRX1
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