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学科主题: 口腔医学
题名:
A 115-bp MethyLight assay for detection of p16 (CDKN2A) methylation as a diagnostic biomarker in human tissues
作者: Zhou, Jing1; Cao, Jie2; Lu, Zheming1; Liu, Hongwei2; Deng, Dajun1
刊名: BMC MEDICAL GENETICS
发表日期: 2011-05-13
DOI: 10.1186/1471-2350-12-67
卷: 12
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Genetics & Heredity
研究领域[WOS]: Genetics & Heredity
关键词[WOS]: ORAL EPITHELIAL DYSPLASIA ; DE-NOVO METHYLATION ; CPG ISLAND ; BARRETTS-ESOPHAGUS ; PROMOTER HYPERMETHYLATION ; MALIGNANT-TRANSFORMATION ; NEOPLASTIC PROGRESSION ; DNA ; ASSOCIATION ; VALIDATION
英文摘要:

Background: p16 Methylation is a potential biomarker for prediction of malignant transformation of epithelial dysplasia. A probe-based, quantitative, methylation-specific PCR (MSP) called MethyLight may become an eligible method for detecting this marker clinically. We studied oral mucosa biopsies with epithelial dysplasia from 78 patients enrolled in a published 4-years′ followup cohort, in which cancer risk for patients with p16 methylation positive dysplasia was significantly higher than those without p16 methylation (by 150-bp MSP and bisulfite sequencing; +133 similar to +283, transcription starting site, +1). The p16 methylation status in samples (N = 102) containing sufficient DNA was analyzed by the 70-bp classic (+238 similar to +307) and 115-bp novel (+157 similar to +272) MethyLight assays, respectively.

Results: p16 Methylation was detectable in 75 samples using the classic MethyLight assay. The methylated-p16 positive rate and proportion of methylated-p16 by the MethyLight in MSP-positive samples were higher than those in MSP-negative samples (positive rate: 37/44 vs. 38/58, P=0.035, two-sided; proportion [median]: 0.78 vs. 0.02, P <0.007). Using the published results of MSP as a golden standard, we found sensitivity, specificity, and accuracy for this MethyLight assay to be 70.5%, 84.5%, and 55.0%, respectively. Because amplicon of the classic MethyLight procedure only partially overlapped with the MSP amplicon, we further designed a 115-bp novel MethyLight assay in which the amplicon on the sense-strand fully overlapped with the MSP amplicon on the antisense-strand. Using the 115-bp MethyLight assay, we observed methylated-p16 in 26 of 44 MSP-positive samples and 2 of 58 MSP-negative ones (P = 0.000). These results were confirmed with clone sequencing. Sensitivity, specificity, and accuracy using the 115-bp MethyLight assay were 59.1%, 98.3%, and 57.4%, respectively. Significant differences in the oral cancer rate were observed during the followup between patients (>= 60 years) with and without methylated-p16 as detected by the 115-bp MethyLight assay (6/8 vs. 6/22, P = 0.034, two-sided).

Conclusions: The 115-bp MethyLight assay is a useful and practical assay with very high specificity for the detection of p16 methylation clinically.

语种: 英语
所属项目编号: 434 ; Z090507017709016 ; 2006AA020902 ; 2006AA02A402
项目资助者: Capital Program for Development of Health Science ; Beijing Science and Technology Commission ; Key Technologies Research &amp ; Development Programs (863)
WOS记录号: WOS:000291947000001
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/60311
Appears in Collections:北京大学口腔医学院_期刊论文

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作者单位: 1.Peking Univ, Key Lab Carcinogenesis & Translat Res, Minist Educ, Dept Aetiol,Canc Hosp & Inst, Beijing 100142, Peoples R China
2.Peking Univ, Dept Oral Med, Sch Stomatol, Beijing 100081, Peoples R China

Recommended Citation:
Zhou, Jing,Cao, Jie,Lu, Zheming,et al. A 115-bp MethyLight assay for detection of p16 (CDKN2A) methylation as a diagnostic biomarker in human tissues[J]. BMC MEDICAL GENETICS,2011,12.
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