5－氮杂－2’－脱氧胞苷对胰腺癌细胞PANC1的组织因子途径抑制物－2表达及CpG岛甲基化的影响  

Effect of 5-Aza-dC on the methylation and expression of TFPI-2 gene in pancreatic cancer PANC1 cell line

作　　者：汤志刚[1] 郑浩黄 强陈炯 

机构地区：[1]安徽医科大学附属省立医院普外科,合肥230001

出　　处：《中华胰腺病杂志》2012年第2期92-94,共3页Chinese Journal of Pancreatology

基　　金：国家自然科学基金（81071734）

摘　　要：目的探讨甲基化酶抑制剂5－氮杂-2’－脱氧胞苷（5-Aza—dC）对胰腺癌细胞系PANC1的抑癌基因组织因子途径抑制物2（tissuefactorpathwayinhibitor2，TFPI-2）甲基化水平及基因表达的影响。方法应用1×10^-7、5×10^-7、1×10μmol／L的5-Aza—dC处理胰腺癌细胞系PANC1。用甲基化特异性PCR（MSP）、RT—PCR及蛋白质印迹法检测细胞TFPI-2基因的甲基化状态、mRNA及蛋白的表达。结果未经5-Aza-dC处理的PANC1细胞的TFPI-2基因CpG岛为完全甲基化，无TFPI-2mRNA及蛋白的表达。1×10^-7、5×10^-7、1×10^-6mol／L的5-Aza-dC处理后，PANC1细胞的TFPI-2基因CpG岛甲基化逆转，从不完全甲基化到完全非甲基化；TFPI-2mRNA相对表达量分别为0．211±0．087、0．327±0．068、0．609±0．017；TFP1-2蛋白相对表达量为0．429±0．121、0．675±0．044、1．132±0．124，呈剂量依赖性增加（P〈0．05）。结论胰腺癌细胞系PANC1的TFPI-2启动子高甲基化可能是导致该基因表达下调甚至失活的主要原因。5-Aza—dC能够逆转其高甲基化状态，并诱导TFPI-2mRNA及蛋白重新表达。Objectives To investigate the effects of 5-aza-2-deoxycytidine（5-Aza-dC）, a methylation inhibitor, on the expression and methylation of tissue factor pathway Inhibitor （TFPI-2） gene in PANC1 cell line of pancreatic cancer. Methods PANC1 cell lines were treated with different dosages of 5-Aza-dC （1 ×10^-7, 5 ×10^-7, 1 × 10^-6 mol/L）. The status of TFPI-2 methylation and expressions of TFPI-2 mRNA and protein were determined by MSP, RT-PCR, and Western blot. Results TFPI-2 gene CpG island was completely methylated, and there was no expression of TFPI-2 mRNA and protein without 5-Aza-dC treatment. After treatment with different dosages of 5-Aza-dC（ 1× 10^-7, 5 ×10^-7, 1× 10^ -6 mol/L）, TFPI-2 gene CpG hypermethylation was reversed from incomplete methylated to complete non-methylated. The relative expressions of TFPI-2 mRNA were 0.211 ±0. 087, 0. 327 ±0. 068, 0. 609 ±0. 017 ; and the relative expressions of TFPI-2 protein were 0.429 ±0. 121, 0.675± 0.044, 1. 132 ±0. 124 in a dose-dependent manner （P 〈0.05）. Conclusions The hypermethylation of promoter region may be the primary reason for TFPI-2 gene expression down-regulation and inactivation. 5-Aza-dC may reverse the hypermethylation of TFPI-2 gene, and induce the re-expression of TFPI-2 mRNA and protein.

关 键 词：胰腺肿瘤 甲基化 组织因子途径抑制物2 5-氮杂-2’-脱氧胞苷 

分 类 号：R735.9[医药卫生—肿瘤]