Highly sensitive determination of the methylated p16 gene in cancer patients by microchip electrophoresis.
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.
Zhou
Xiao-Mian
XM
Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, P.R. China.
Shao
Shu-Juan
SJ
Xu
Guan-Dong
GD
Zhong
Run-Tao
RT
Liu
Da-Yu
DY
Tang
Jian-Wu
JW
Gao
Yan-Ning
YN
Cheng
Shu-Jun
SJ
Lin
Bing-Cheng
BC
D000008:Abdominal Neoplasms / Q000097:blood / Q000235:genetics; D019175:DNA Methylation; D046209:Electrophoresis, Microchip / Q000295:instrumentation / Q000379:methods*; D005240:Feasibility Studies; D019942:Genes, p16; D006801:Humans; D008175:Lung Neoplasms / Q000097:blood / Q000235:genetics; D009369:Neoplasms / Q000235:genetics*; D019904:Polymethyl Methacrylate; D012680:Sensitivity and Specificity
D019904:Polymethyl Methacrylate