Zhou Xiao-Mian, Shao Shu-Juan, Xu Guan-Dong, Zhong Run-Tao, Liu Da-Yu, Tang Jian-Wu, Gao Yan-Ning, Cheng Shu-Jun, Lin Bing-Cheng
Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, P.R. China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2005 Feb 25;816(1-2):145-51. doi: 10.1016/j.jchromb.2004.11.045.
The p16 tumor suppressor gene is inactivated by promoter region hypermethylation in many types of tumor. Recent studies showed that aberrant methylation of the p16 gene is an early event in many tumors, especially in lung cancer, and may constitute a new biomarker for early detection and monitoring of prevention trials. We detected tumor-associated aberrant hypermethylation of the p16 gene in plasma and tissue DNA from 153 specimens using a modified semi-nested methylation-specific PCR (MSP) combining plastic microchip electrophoresis or slab gel electrophoresis, respectively. Specimens were from 79 lung cancer patients, 15 abdominal tumor patients, 30 positive controls and 30 negative controls. The results showed that the positive rate obtained by microchip electrophoresis was more than 26.6% higher and the same specificity was kept when compared with slab gel electrophoresis. The microchip electrophoresis can rapidly and accurately analyze the PCR products of methylated DNA and obviously improve the positive rate of diagnosis of cancer patients when compared with gel electrophoresis. This method with the high assay sensitivity might be used for detection of methylation of p16 gene and even to facilitate early diagnosis of cancer patients.
p16肿瘤抑制基因在多种肿瘤中因启动子区域高甲基化而失活。近期研究表明，p16基因的异常甲基化是许多肿瘤尤其是肺癌中的早期事件，可能构成早期检测及预防试验监测的一种新生物标志物。我们分别使用结合塑料微芯片电泳或平板凝胶电泳的改良半巢式甲基化特异性PCR（MSP），检测了153份标本的血浆和组织DNA中与肿瘤相关的p16基因异常高甲基化。标本来自79例肺癌患者、15例腹部肿瘤患者、30例阳性对照和30例阴性对照。结果显示，与平板凝胶电泳相比，微芯片电泳获得的阳性率高出26.6%以上，且特异性相同。微芯片电泳能够快速、准确地分析甲基化DNA的PCR产物，与凝胶电泳相比，可显著提高癌症患者的诊断阳性率。这种具有高检测灵敏度的方法可用于检测p16基因的甲基化，甚至有助于癌症患者的早期诊断。
