Quantitative detection of promoter hypermethylation of multiple genes in the tumor, urine, and serum DNA of patients with renal cancer
Quantitative detection of promoter hypermethylation of multiple genes in the tumor, urine, and serum DNA of patients with renal cancer
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DOI：
10.1158/0008-5472.can-04-0799
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发表时间：
2004-08-01
期刊：
影响因子：
11.2
通讯作者：
Sidransky, D
中科院分区：
文献类型：
--
作者：
Hoque, MO;Begum, S;Sidransky, D
几种已知或推定的肿瘤抑制基因的异常启动子高甲基化在人类癌症的发病机制中经常发生，并且是癌症检测的有希望的标志物。我们研究了检测肾癌患者尿液和血清样本中异常 DNA 甲基化的可行性。我们通过定量荧光实时 PCR 检查了 17 名原发性肾癌患者的肿瘤以及匹配的尿液和血清 DNA 中 9 个基因启动子（CDH1、APC、MGMT、RASSF1A、GSTP1、p16、RAR-β2 和 ARF）的异常甲基化。还对肾癌患者的另外 9 个尿液样本（总共 26 个）和 I 血清样本（总共 18 个）进行了测试。 91 名非泌尿生殖系癌症患者的尿液和 30 名年龄匹配的非癌症个体的血清用作对照。在 17 个原发性肿瘤中的 16 个（94%）中检测到至少两个所研究基因的启动子高甲基化。尿液和血清 DNA 中的异常甲基化通常伴随着匹配的肿瘤样本中的甲基化。来自 91 名没有泌尿生殖系统癌症证据的对照受试者的尿液样本显示，MGMT、GSTP1、p16 和 ARF 基因没有甲基化，而在少数对照受试者中检测到低水平的 RAR-beta2、RASSF1A、CDH1、APC 和 TIMP3 甲基化。总体而言，癌症患者的 26 个尿液样本中的 23 个（88%）和 18 个血清样本中的 12 个（67%）至少检测到一种基因甲基化呈阳性。通过结合肾癌患者的尿液或血清分析，17 名患者中有 16 名检测到高甲基化（敏感性为 94%），具有高特异性。我们的研究结果表明，在大多数肾癌患者中可以检测到尿液或血清中的启动子高甲基化。这种非侵入性高通量方法需要在大型研究中进行评估，以评估其在肾癌早期检测和监测中的价值。
Aberrant promoter hypermethylation of several known or putative tumor suppressor genes occurs frequently during the pathogenesis of human cancers and is a promising marker for cancer detection. We investigated the feasibility of detecting aberrant DNA methylation in the urine and serum samples of renal cancer patients. We examined the tumor and the matched urine and serum DNA for aberrant methylation of nine gene promoters (CDH1, APC, MGMT, RASSF1A, GSTP1, p16, RAR-beta2, and ARF) from 17 patients with primary kidney cancer by quantitative fluorogenic real-time PCR. An additional 9 urine samples (total, 26) and I serum sample (total, 18) also were tested from renal cancer patients. Urine from 91 patients without genitourinary cancer and serum from 30 age-matched noncancer individuals were used as controls. Promoter hypermethylation of at least two of the genes studied was detected in 16 (94%) of 17 primary tumors. Aberrant methylation in urine and serum DNA generally was accompanied by methylation in the matched tumor samples. Urine samples from 91 control subjects without evidence of genitourinary cancer revealed no methylation of the MGMT, GSTP1, p16, and ARF genes, whereas methylation of RAR-beta2, RASSF1A, CDH1, APC, and TIMP3 was detected at low levels in a few control subjects. Overall, 23 (88%) of 26 urine samples and 12 (67%) of 18 serum samples from cancer patients were methylation positive for at least one of the genes tested. By combination of urine or serum analysis of renal cancer patients, hypermethylation was detected in 16 of 17 patients (94% sensitivity) with high specificity. Our findings suggest that promoter hypermethylation in urine or serum can be detected in the majority of renal cancer patients. This noninvasive high-throughput approach needs to be evaluated in large studies to assess its value in the early detection and surveillance of renal cancer.