A highly sensitive method for the detection of p16 methylation in the serum of colorectal cancer patients.
A highly sensitive method for the detection of p16 methylation in the serum of colorectal cancer patients.
复制标题
一种检测结直肠癌患者血清中 p16 甲基化的高灵敏度方法。
作者：
G. Nakayama;K. Hibi;H. Nakayama;Y. Kodera;Katsuki Ito;S. Akiyama;A. Nakao
背景
为了早期检测癌症，人们引入了通过甲基化特异性聚合酶链反应（MSP）检测多种恶性肿瘤血清中p16超甲基化的方法。据报道，尝试增加该测定法用于结直肠癌的灵敏度。
材料和方法
DNA样品在被分成10个较小的样品之前进行有限稀释。随后，对这10个样品进行半巢式MSP。与常规MSP相比，有限稀释MSP（LD-MSP）检测甲基化DNA的灵敏度提高了10倍。
结果
在44例结直肠癌患者中，30例（68%）血清DNA中p16基因启动子甲基化异常，而常规MSP检测13例（30%）。作为对照，50例大肠癌患者的血清DNA中相应的肿瘤DNA在p16启动子中没有甲基化，筛选异常甲基化。用LD-MSP和常规MSP检测对照组血清DNA均未发现甲基化。
结论
LD-MSP的高灵敏度使其能够检测血清中比常规MSP更少量的肿瘤DNA。该技术也具有很高的特异性，如果相应的肿瘤不表现出甲基化，则尚未观察到血清中的异常甲基化。这一观察结果支持了LD-MSP可用于临床检测血清中肿瘤DNA的想法。
BACKGROUND
For the purpose of early detection of carcinomas, detection of p16 hypermethylation by methylation-specific polymerase chain reaction (MSP) in the serum of many kinds of malignancies has been introduced. An attempt to increase the sensitivity of this assay is reported for colorectal cancer.
MATERIALS AND METHODS
DNA samples were subjected to limiting dilution before being divided into ten smaller samples. Subsequently, hemi-nested MSP was performed on those ten samples. The limiting dilution-MSP (LD-MSP) provided a 10-fold increase in sensitivity of the detection of methylated DNA compared with conventional MSP.
RESULTS
Of 44 colorectal cancer patients, 30 (68%) exhibited abnormal promoter methylation of p16 in their serum DNA by LD-MSP, while 13 (30%) exhibited it by conventional MSP. As a control, the serum DNA of 50 patients with colorectal carcinomas whose corresponding tumor DNA had no methylation in the p16 promoter was screened for aberrant methylation. No methylation was found in the serum DNA of this control group by LD-MSP or conventional MSP.
CONCLUSION
The high sensitivity of LD-MSP makes it possible to detect smaller amounts of tumor DNA in the serum than conventional MSP. This technique also has great specificity and no abnormal methylation in serum has yet been observed if the corresponding tumor does not exhibit methylation. This observation supports the idea that LD-MSP could be applied in clinical use for the detection of tumor DNA in serum.
高甲基化可以选择性地沉默肿瘤中的个体 p16ink4A 等位基因。
DOI：
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发表时间：
1998
期刊：
Cancer research.
影响因子：
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作者：
Myohanen,SK;Baylin,SB;Herman,JG
通讯作者：
Herman,JG
影响因子：
11.2
作者：
M. Sanchez-Cespedes;M. Esteller;Li Wu;H. Nawroz-Danish;G. Yoo;W. Koch;Jin Jen;J. Herman;D. Sidransky
通讯作者：
M. Sanchez-Cespedes;M. Esteller;Li Wu;H. Nawroz-Danish;G. Yoo;W. Koch;Jin Jen;J. Herman;D. Sidransky
影响因子：
11.2
作者：
M. Esteller;S. Tórtola;Minoru Toyota;G. Capellá;M. Peinado;S. Baylin;James G. Herman
通讯作者：
M. Esteller;S. Tórtola;Minoru Toyota;G. Capellá;M. Peinado;S. Baylin;James G. Herman