免疫沉淀核小体 DNA 是一种提高结肠癌相关 p16 高甲基化血清筛选敏感性的新方法。
Immunoprecipitation of nucleosomal DNA is a novel procedure to improve the sensitivity of serum screening for the p16 hypermethylation associated with colon cancer.
机构信息
Division of Gastroenterology and Hematology/Oncology, Department of Medicine, Asahikawa Medical College, 2-1-1-1, Midorigaoka-Higashi, Asahikawa, Hokkaido 078-8510, Japan.
出版信息
Cancer Epidemiol. 2010 Apr;34(2):194-9. doi: 10.1016/j.canep.2010.01.004. Epub 2010 Feb 2.
BACKGROUND
We developed a novel method of methylation-specific PCR (MSP) using immunoprecipitation with anti-histone antibody (IP-MSP) to efficiently detect serum methylated DNA tightly bound to de-acetylated histones.
MATERIALS AND METHODS
The detection limit of IP-MSP for p16 methylation was determined with a standard made by cell line (SKCO-1) lysate. p16 methylation of tumor and/or serum of 51 colorectal cancers and 10 adenoma patients, and 10 healthy volunteers was detected with conventional MSP or IP-MSP.
RESULTS
IP-MSP detected p16 methylation from 0.5pg/mul of the cell lysate. The sensitivity of IP-MSP for detecting serum p16 methylation in 27 patients with tumors characterized by p16 methylation was significantly higher than that with conventional method (81% versus 59%), particularly in Stage II patients (91% versus 45%). IP-MSP detected no p16 hypermethylation in sera of adenoma patients and volunteers.
CONCLUSIONS
IP-MSP is thus considered to be a promising procedure to detect serum methylated DNA in colorectal cancer patients.
背景
我们开发了一种新的甲基化特异性聚合酶链反应（MSP）方法，使用抗组蛋白抗体免疫沉淀（IP-MSP），以有效地检测与去乙酰化组蛋白紧密结合的血清甲基化 DNA。
材料和方法
使用细胞系（SKCO-1）裂解物制成的标准品，确定 IP-MSP 检测 p16 甲基化的检测限。使用常规 MSP 或 IP-MSP 检测 51 例结直肠癌和 10 例腺瘤患者的肿瘤和/或血清中的 p16 甲基化以及 10 名健康志愿者的 p16 甲基化情况。
结果
IP-MSP 从 0.5pg/mul 的细胞裂解物中检测到 p16 甲基化。在 27 例 p16 甲基化特征的肿瘤患者中，IP-MSP 检测血清 p16 甲基化的灵敏度明显高于常规方法（81%对 59%），尤其是在 II 期患者中（91%对 45%）。IP-MSP 未在腺瘤患者和志愿者的血清中检测到 p16 过度甲基化。
结论
因此，IP-MSP 被认为是一种有前途的方法，可用于检测结直肠癌患者的血清甲基化 DNA。
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