胰腺癌细胞株原钙黏附蛋白8基因的甲基化状态  

Methylation of PCDH8 in pancreatic carcinoma cell lines

作　　者：吕顺莉[1] 高军[1] 杜奕奇[1] 黄浩杰[1] 王小玮[1] 金晶[1] 龚燕芳[1] 张玲[1] 李兆中[1] 

机构地区：[1]第二军医大学长海医院消化内科,上海200433

出　　处：《中华胰腺病杂志》2010年第3期190-192,共3页Chinese Journal of Pancreatology

摘　　要：目的 分析原钙黏附蛋白8（protocadherin 8,PCDH8）基因在胰腺癌细胞株的甲基化状态.方法 抽提6株胰腺癌细胞株PANC1、ASPC1、BxPC3、CFPAC、PaTu8988、SW1990和2例正常胰腺组织的总RNA,以甲基化特异性PCR（MSP）法检测PCDH8甲基化情况.应用DNA甲基化转移酶（DNMT）抑制剂5-氮杂2＇-脱氧胞苷（5-Aza-dC）处理6株胰腺癌细胞株,采用实时定量PCR法检测处理前后细胞的PCDH8 mRNA表达.结果 2例正常胰腺组织PCDH8基因未发生甲基化,PANC1、BxPC3、CFPAC胰腺癌细胞株PCDH8基因部分甲基化,而PaTu8988、ASPC1、SW1990细胞完全甲基化.PCDH8mRNA在PANC1、SW1990、PaTu8988胰腺癌细胞株中有表达,表达的相对值（RQ）分别为1.576±0.648、0.013±0.008、0.002±0.001;BxPC3、CFPAC、ASPC1细胞株无PCDH8 mRNA表达.5-Aza-dC处理后,胰腺癌细胞株PANC1、ASPC1、BxPC3、CFPAC、PaTu8988、SW1990均有PCDH8 mRNA表达,表达量较处理前明显升高,相对表达量分别为7.463±2.628、10.696±1.539、7.852±2.762、421.815±1.493、118.595±4.089、6.690±1.884.结论 PCDH8基因启动子高甲基化是导致该基因在胰腺癌细胞株表达下降的主要原因之一.Objective To investigate the methylation status of PCDH8 gene in pancreatic carcinoma.Methods Methylation of PCDH8 gene in 2 samples of normal pancreatic tissues and 6 pancreatic carcinoma cell lines （PANC1, ASPC1, BxPC3, CFPAC, PaTu8988 and SW1990） was detected by the methylationspecific PCR （MSP） method. The expression of PCDH8 mRNA was detected with 5-Aza-2-deoxycytidine （5-Aza-dC） treatment, a kind of DNA methyltransferase （DNMT） inhibitor in 6 pancreatic carcinoma cell lines by real-time-PCR. Results The methylation of PCDH8 gene was not detected in normal tissues, while it was partially methylated in PANC1, BxPC3, CFPAC and it was totally methylated in PaTu8988, ASPC1, SW1990.PCDH8 mRNA was expressed in PANC1, SW1990, PaTu8988 and the relative quantities of mRNA expression （RQ） were 1.576 ± 0.648, 0.013 ± 0.008, 0.002 ± 0.001; PCDH8 mRNA was not expressed in BxPC3,CFPAC, ASPC1. After 5-Aza-dC treatment, PCDH8 mRNA was expressed in PANC1, ASPC1, BxPC3,CFPAC, PaTu8988, SW1990 and the relative quantities of mRNA expression all significantly increased, and they were 7. 463 ± 2.628, 10. 696 ± 1.539, 7.852 ± 2.762,421.815 ± 1.493, 118.595 ± 4.089, 6.690 ±1.884. Conclusions The methylation of PCDH8 gene may be the major mechanism of down-regulated expression of PCDH8 gene in pancreatic carcinoma.

关 键 词：胰腺肿瘤 甲基化 聚合酶链反应 原钙黏附蛋白8 

分 类 号：R735.9[医药卫生—肿瘤]