乳腺癌bcl-2甲基化的检测及与表达、预后的关系

Detection of bcl-2 methylation and the relationship between bcl-2 methylation and expression, prognostic factors in breast cancer

目的:建立Bcl-2基因甲基化特异的PCR(MSP)检测方法,并探讨乳腺癌bcl-2甲基化与蛋白表达、 预后因素(肿瘤大小,淋巴结转移,增殖细胞核抗原PCNA,雌、孕激素受体ER、PR情况)的关系。方法:设计bcl-2基 因MSP引物,采用MSP方法检测54例乳腺癌bcl-2基因5’端启动子CpG岛甲基化状态。采用免疫组化S-P法检 测54例乳腺癌bcl-2、PCNA、ER、PR的表达。结果:乳腺癌bcl-2甲基化率为29．6％。Bcl-2甲基化与其蛋白表达 之间呈显著负相关(P<0．01)。Bcl-2甲基化率高与不良的预后因素(PCNA标记指数LI高、ER-和PR-)显著相关 (P<0．01)。结论:该研究建立bcl-2基因MSP检测方法,MSP扩增和测序结果证实,bcl-2基因MSP引物设计是合 理的。bcl-2甲基化有可能成为乳腺癌预后不良的分子检测指标。

AIM： To establish methylation - specific PCR （MSP） method for detecting bel - 2 gene, and to study the relationship between bel - 2 methylation and expression, prognostic factors in breast cancer. METHODS： The primer of bel - 2 gene for MSP was designed. The methylations in CpG island of bel - 2 gene in 54 cases of breast cancer were detected by using MSP. The expressions of bcl - 2, PCNA, ER and PR in 54 eases of breast cancer were detected by using SP technique. RESULTS： The overall positive rate of bcl - 2 methylation was 29.6% in breast cancer. There was a significant negative correlation between the methylation of bcl - 2 and the expression of bel - 2 （ P 〈 0.01 ）. The methylation of bel - 2 coincided with those bad prognostic factors such as high PCNA label index （LI）, ER - and PR - （ P 〈 0.01 ）. CONCLUSIONS： This study established the MSP method for detecting bel - 2 gene. The results of MSP and sequence analysis testified that the design of the MSP primer of bel - 2 gene in this study was successful. The methylation of bel - 2 would become the marker indicating bad prognosis of breast cancer.