血浆DNA甲基化检测在宫颈疾病诊断中的应用

Application of plasma DNA methylation determination in diagnosis of cervix diseases

目的对宫颈疾病患者血浆APC和RASSF1A基因启动子甲基化水平进行定量检测,探讨其在宫颈疾病诊断中的意义。方法收集34例宫颈低度病变(CINⅠ期)、43例宫颈高度病变(包含CINⅡ、CINⅢ期)、41例宫颈癌患者治疗前血浆及25例健康妇女血浆。另收集31例与血浆对应的宫颈组织标本。用双重实时荧光定量甲基化特异性PCR(qMSP)技术检测APC和RASSF1A基因启动子甲基化水平并进行统计学分析。结果宫颈疾病患者血浆中APC和RASSF1A甲基化水平与肿瘤组织有较好的一致性,且肿瘤组织水平高于血浆水平(Z分别为-2.236和-3.228,P均<0.05)。宫颈癌组血浆APC与RASSF1A甲基化率均高于低度病变组(P均<0.05);宫颈高度病变组血浆RASSF1A甲基化率也高于低度病变组,差异有显著性(P<0.05)。血浆APC和RASSF1A甲基化阳性结果在低度病变组、高度病变组和宫颈癌组均无显著性差异(P>0.05)。结论血浆DNA甲基化水平能反映宫颈癌的发生、发展变化,在宫颈癌的早期诊断中可能有重要价值。

Objective To determine quantitatively the levels of plasma adenomatous polyposis coli （APC） and Ras association domain family 1A （RASSF1A） gene promoter DNA methylation in the patients with cervical disease, and evaluate their significance in diagnosis of cervix diseases. Methods Preoperative plasma samples were collected from 34 patients with low-grade cervical lesion （CIN I ） , 43 with high-grade lesion （ CIN II and III ） and 41 with cervical cancer. In addition, 25 plasma samples from healthy women and 31 cervical tissue specimens were also collected. The DNA methylation levels of APC and RASSF1A gene promoter were determined by duplex real-time fluorescence quantitative methylation-specific PCR（qMSP） technology. The related statistical analysis was performed. Results The APC and RASSFIA methylation levels in plasma were positively correlated with those in tumor tissue in the patients with cervical disease, and the levels in tumor tissue were significantly higher than those in plasma （ Z = - 2. 236 and - 3. 228, respectively, P 〈 0.05 ）. The plasma APC and RASSF1A methylation rates in both cervical cancer and high-grade lesion group were significantly higher than those in low-grade lesion group （P 〈 0.05）. However, there was no significant difference of plasma APC and RASSF1A methylation rates between the low-grade lesion and the healthy control group（ P 〉 0.05 ）. Conclusion The plasma methylation levels of APC and RASSF1A gene promoter were able to reflect the development changes of cervical cancer, which may have important value in the early diagnosis of cervical cancer.