摘要
目的探讨关节软骨中组织金属蛋白酶抑制因子-3(TIMP-3)基因启动子区甲基化与其蛋白表达的相关性,并分析TIMP-3基因CpG岛异常甲基化与骨关节炎(OA)的关联性。方法应用甲基化特异性的聚合酶链反应(MSP)技术和免疫组织化学SP法分别检测14例健康人的正常关节软骨细胞和35例骨关节炎(OA)患者软骨细胞TIMP-3基因启动子区甲基化和蛋白表达情况。结果 OA患者和健康人关节软骨中TIMP-3基因启动子区均有甲基化修饰,其阳性率分别为74.3%(26/35)和35.7%(5/14),OA组TIMP-3基因启动子区甲基化率明显高于健康组(P<0.05)。14例健康人中,软骨细胞TIMP-3蛋白表达阳性10例(71.4%),而35例OA患者中,软骨细胞TIMP-3蛋白表达阳性11例(31.4%)。24例OA患者软骨细胞蛋白表达阴性的标本中,TIMP-3启动子区甲基化阳性21例(87.5%);26例TIMP-3启动子区甲基化阳性的标本中,蛋白表达阴性21例(80.8%),TIMP-3启动子区甲基化与蛋白表达呈显著负相关(P<0.05)。结论启动子区CpG岛高甲基化是OA患者关节软骨细胞TIMP-3表达失活的主要机制之一,其可能参与了OA的发生和发展。
Objective To investigate the relationship between the methylation of tissue inhibitor of metalloproteinase-3 (TIMP-3) gene promoter and its protein expression, to study the potential role of hyper-methylation of TIMP-3 gene in the development of osteoarthritis. Methods The CpG islands methylation levels of TIMP-3 promoter in joint cartilage cells were studied by methylation-specific PCR(MSP). TIMP-3 protein expression in joint cartilage cells were detected by immunohistochemical method in 35 osteoarthritis (OA) patients and 14 control health people. Results The positive percent of TIMP-3 promoter methylation levels in healthy people and OA patients were 35.7%(5/14) and 74.3%(26/35) respectively. The positive percent of TIMP-3 protein expression in OA patients were higher than healthy people (P〈 0.05); In cartilage cells, the positive percent of TIMP-3 protein expression in healthy people and OA patients were 71.4%(10/14) and 31.4%(11/35) respectively. In 24 cases of OA samples with negative cartilage cells protein expres- sion, 21 cases (87.5%)was found with positive TIMP-3 promoter methylation, and in 26 cases of patients with positive TIMP-3 promoter methylation, protein expression was negative in 21 cases (80.8%);Promoter methylation and TIMP-3 protein expression level showed significantly negative correlation. Conclusion TIMP-3 promoter is hyper-methlated in joint cartilage cells of OA patients. The hyper-methylation affects the protein expression of TIMP-3, which may play a crtical role in the occurrence and development of OA.
出处
《中国当代医药》
2014年第2期4-7,共4页
China Modern Medicine
基金
湖南省科技计划一般项目(2010FJ6004)
关键词
骨关节炎
甲基化
TIMP-3启动子
CPG岛
Osteoarthritis
Methylation
Tissue inhibitor of metalloproteinase-3 gene promoter
CpG islands