Combined CADM1 and MAL promoter methylation analysis to detect (pre-)malignant cervical lesions in high-risk HPV-positive women
Combined CADM1 and MAL promoter methylation analysis to detect (pre-)malignant cervical lesions in high-risk HPV-positive women
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DOI：
10.1002/ijc.25890
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发表时间：
2011-11-01
影响因子：
6.4
通讯作者：
Steenbergen, Renske D. M.
中科院分区：
文献类型：
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作者：
Overmeer, Renee M.;Louwers, Jacqueline A.;Steenbergen, Renske D. M.
鉴于与细胞学检测相比，高危人乳头瘤病毒 (hrHPV) 检测对高级宫颈病变的特异性较低，因此需要对 hrHPV 检测呈阳性的女性进行额外的分类检测，以检测高级宫颈病变。在这里，我们测试了细胞粘附分子 1 (CADM1) 和 T 淋巴细胞成熟相关蛋白 (MAL) 的联合甲基化分析是否可以作为这样的分诊标志物，这两种蛋白都在功能上参与宫颈癌的发生。对 261 个宫颈组织标本（从非肿瘤到癌）应用了 4 种定量甲基化特异性 PCR (qMSP)，其中 2 种针对 CADM1（M12 和 M18 区域）和 MAL（M1 和 M2 区域）。当组合 qMSP 并考虑组合中至少一个 qMSP 的阳性时，通过将单个 CADM1 标记物与单个 MAL 标记物组合，可以获得宫颈上皮内瘤变 3 级 (CIN3) 病变 (97%) 和鳞状细胞癌和腺癌 (99%) 的最高阳性率。随后对 70 份 GP5+/61-PCR hrHPV 阳性刮片进行的 qMSP 分析显示，由 CADM1-M18 和 MAL-M1 组成的双标记物组合最具辨别力，检测出 90% 的 CIN3 女性 (n = 30)，而在无宫颈疾病的女性 (n = 40) 中只有 13.5% 的结果呈阳性。最后，我们对 79 名到阴道镜门诊就诊的女性进行了 hrHPV GP5+/6+-PCR 检测，然后进行了 CADM1-M18/MAL-M1 甲基化分析。 hrHPV 检测显示 CIN3+ 的敏感性为 97%，特异性为 33%。对 hrHPV 阳性女性进行的额外 CADM1-M18/MAL-M1 甲基化分析将特异性提高到 78%，敏感性为 70%。总之，甲基化标记物组 CADM1-M18 和 MAL-M1 可作为 hrHPV 阳性女性的替代分子分类工具。
Given the lower specificity for high-grade cervical lesions of high-risk human papillomavirus (hrHPV) testing compared to cytology, additional triage testing for hrHPV test-positive women is needed to detect high-grade cervical lesions. Here, we tested whether combined methylation analysis for cell adhesion molecule 1 (CADM1) and T-lymphocyte maturation associated protein (MAL), both functionally involved in cervical carcinogenesis, could serve as such a triage marker. Four quantitative methylation-specific PCRs (qMSP), two for CADM1 (regions M12 and M18) and MAL (regions M1 and M2) each, were applied to 261 cervical tissue specimens ranging from no neoplasia to carcinoma. When qMSPs were combined and positivity for at least one of the qMSPs in the combination was taken into account, the highest positivity rates for cervical intraepithelial neoplasia grade 3 (CIN3) lesions (97%) and squamous cell-and adeno-carcinomas (99%) were obtained by combining a single CADM1 marker with a single MAL marker. Subsequent qMSP analysis of 70 GP5+/61-PCR hrHPV-positive scrapings revealed that a two-marker panel consisting of CADM1-M18 and MAL-M1 was most discriminative, detecting 90% of women with CIN3 (n = 30), whereas it showed a positive result in only 13.5% of women without cervical disease (n = 40). Finally, we applied hrHPV GP5+/6+-PCR testing followed by CADM1-M18/MAL-M1 methylation analysis to a cohort of 79 women visiting the outpatient colposcopy clinic. hrHPV testing revealed a sensitivity of 97% and a specificity of 33% for CIN3+. Additional CADM1-M18/MAL-M1 methylation analysis on the hrHPV-positive women increased the specificity to 78% with a sensitivity of 70%. In conclusion, the methylation marker panel CADM1-M18 and MAL-M1 may serve as an alternative molecular triage tool for hrHPV-positive women.