尖锐湿疣组织生存素基因外显子1甲基化的检测及意义

目的研究尖锐湿疣(CA)组织生存素基因外显子1区CpG岛甲基化与其基因表达的关系,探讨生存素在CA组织中的表达及其调控机制.方法采用免疫组化方法检测30例CA皮损、10例正常宫颈、10例正常包皮组织中生存素的表达;运用RT-PCR方法检测生存素的转录水平;用甲基化特异性聚合酶链反应(MSP)检测生存素外显子1 CpG岛甲基化状态.结果生存素蛋白和mRNA在CA组的阳性表达率分别为90%、93.3%,而正常对照组分别为5%、5%,两组间差异均有统计学意义(x2=35.187、38.437,P值均<0.01);生存素基因外显子1 CpG岛去甲基化率在30例CA组织为86.7%.而在20例正常组织为15%,组间比较差异有统计学意义(x2=10.865,P<0.01),且生存素外显子1 CpG岛去甲基化和mRNA表达呈显著正相关(x2=13.929,P=0.014).结论生存素在CA的表达可能与其外显子1区CpG岛去甲基化有关,并可能在CA的发生过程中起一定作用.

Detection of methylation in exon 1 of the survivin gene in condyloma acuminatum (CA) tissue and its significance

Objective To investigate the relationship between the expression of the survivin gene and CpC methylation in exon 1 of the survivin gene in CA tissue, and to study the expression of survivin protein in CA tissue and its modulation mechanism. Methods Tissue samples were obtained from the CA lesions of 30 patients, normal cervix of 10 female controls, and normal foreskin of 10 male controls. Immunohistochemistry was carried out to detect the expression of survivin protein in these specimens, RT-PCR to measure the mRNA expression of survivin, and methylation specific PCR (MSP) to analyze the methylation status of CpG island in the survivin gene exon 1. Results The positivity rate of survivin protein and mRNA was 90% (27/30) and 93.3% (28/30) in CA tissue specimens, respectively, 5% (1/20) and 5% (1/20) in control tissue specimens, respectively; there was a significant difference between the two groups of specimens in both the parameters (x2 = 35.187, 38.437, both P < 0.01). The demethylation of CpG island in the survivin gene exon 1 was observed in 86.7% (26/30) of the CA tissue specimens and in 15% (3/20) of the control tissue specimens (x2 = 10.865, P < 0.01). There was a positive correlation between the demethylation status of CpG island in exon 1 and the mRNA expression of survivin gene (x2 = 13.929, P = 0.014). Conclusions The expression of survivin protein in CA tissues might be associated with the demethylation of CpG island in exon 1 of the survivin gene, and may play a certain role in the development of CA.