Quantitative methylation-specific PCR for the detection of aberrant DNA methylation in liquid-based pap tests
Quantitative methylation-specific PCR for the detection of aberrant DNA methylation in liquid-based pap tests
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DOI：
10.1002/cncr.23258
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发表时间：
2008-02-25
影响因子：
3.4
通讯作者：
Gustafson, Karen S.
中科院分区：
文献类型：
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作者：
Kahn, Steven L.;Ronnett, Brigitte M.;Gustafson, Karen S.
背景。选择性肿瘤抑制基因的异常启动子甲基化已在鳞状上皮内病变（SIL）和侵袭性宫颈癌中被检测到。鉴定可以区分高级别SIL （HSIL）和低级别SIL (LSIL)，以及上皮内病变或恶性肿瘤（NILM）残留液基巴氏试验（Pap）细胞学阴性的基因甲基化谱，可能是宫颈癌筛查的辅助检测方法。利用实时定量甲基化特异性聚合酶链反应（QMSP），作者分析了活检证实的HSIL和LSIL以及NILM残留液体巴氏试验中DAPK1、IGSF4、SPARC和TFP12启动子甲基化的频率和相对水平。使用内参基因ACTB计算每个基因的甲基化百分比（%M）。每个样本的累积甲基化评分，定义为所有4个基因的%M之和，用于分析基因组合。对于所分析的每个基因，与NILM/LSIL联合样本相比，HSIL中甲基化的频率和相对水平均有所增加。HSIL样本的累积甲基化评分明显更高(P
BACKGROUND. Aberrant promoter methylation of selective tumor suppressor genes has been detected in squamous intraepithelial lesions (SIL) and invasive cervical cancer. Identification of methylation profiles of genes that can distinguish high-grade SIL (HSIL) from low-grade SIL (LSIL), and cytologically negative for intraepithelial lesion or malignancy (NILM) residual liquid-based Papanicolaou (Pap) tests may be potentially useful as an ancillary test for cervical cancer screening.METHODS. Using real-time quantitative methylation-specific polymerase chain reaction (PCR) (QMSP), the authors analyzed the frequency and relative level of promoter methylation for DAPK1, IGSF4, SPARC, and TFP12 in biopsy-confirmed HSIL and LSIL, and NILM residual liquid-based Pap tests. The percentage of methylation (%M) for each gene was calculated using the reference gene, ACTB. The cumulative methylation score for each sample, defined as the sum of %M of all 4 genes, was used to analyze the genes in combination.RESULTS. For each gene analyzed the frequency and relative level of methylation were increased in HSIL compared with combined NILM/LSIL samples. The cumulative methylation scores were significantly higher in HSIL samples (P