Quantitative DNA methylation analysis detects cervical intraepithelial neoplasms type 3 and worse

Abstract

BACKGROUND: DNA methylation may be used a potential biomarker for detecting cervical cancer. The authors of this report used quantitative methylation analysis of 4 genes in a full spectrum of cervical lesions to test its potential clinical application. METHODS: This hospital-based, retrospective, case-control study was conducted in 185 patients and included patients who had a normal uterine cervix (n ¼ 53), cervical intraepithelial neoplasm type 1 (CIN1) (n ¼ 37), CIN2 (n ¼ 22), CIN3 (n ¼ 24), carcinoma in situ (CIS) (n ¼ 22), squamous cell carcinoma (SCC, n ¼ 20), and adenocarcinoma (AC) (n ¼ 7). Methylation levels of the genes sex-determining region Y, box 1 (SOX1); paired box gene 1 (PAX1); LIM homeobox transcription factor 1a (LMX1A), and NK6 transcription factor-related locus 1 (NKX6-1) were determined by using real-time methylation-specific polymerase chain reaction (PCR) amplification. Cutoff values of the percentage of methylation reference (PMR) for different diagnoses were determined to test the sensitivity and specificity and to generate receiver operating characteristic (ROC) curves. Two-sided Mann-Whitney U tests were used to test differences in PMR between groups. RESULTS: The PMRs of the 4 genes were significantly higher in CIN3 and worse (CIN3þ) lesions than the PMRs in specimens of normal cervix and CIN1 or CIN2 (P < .001). ROC curve analysis demonstrated that the sensitivity, specificity, and accuracy for detecting CIN3þ lesions were 0.88, 0.82, and 0.95, respectively, for SOX1; 0.78, 0.91, and 0.89, respectively, for PAX1; 0.77, 0.88, and 0.90, respectively, for LMX1A; and 0.93, 0.97, and 0.97, respectively, for NKX6-1. CONCLUSIONS: The current results indicated that quantitative PCR-based testing for DNA methylation of 4 genes holds great promise for cervical cancer screening and warrants further population-based studies using standardized DNA methylation testing. Cancer 2010;116:4266-74.

HPV33 DNA methylation measurement improves cervical pre-cancer risk estimation of an HPV16, HPV18, HPV31 and \textit{EPB41L3} methylation classifier

“…We and other research groups have observed that measurement of differential methylation of certain HPV CpG sites is useful for stratifying HPV type-specific risk [8,[11][12][13]19]. Additionally, methylation of CpGs in the promoters or introns of human genes has shown some value [5,6,9,14,20]. In earlier work we demonstrated that combining the measurement of human and HPV methylation might produce a better classifier than either alone [1].…”

PAX1 and SOX1 methylation as an initial screening method for cervical cancer: a meta-analysis of individual studies in Asians

“…Among these altered and methylated genes, the paired boxed gene 1 (PAX1) and sex determining region Y-box 1 (SOX1) were both highlighted (12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26). Numerous studies have documented the promise of PAX1 or SOX1 DNA methylation in distinguishing patients with reactive atypia or CIN3+ (cervical intraepithelial neoplasia type III or worse) lesions from normal uterine cervixes (12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23). In order to make a comparison of the accuracy of DNA methylation and HPV DNA testing, we performed a comprehensive meta-analysis and evaluated the diagnostic performance of PAX1 and SOX1 methylation for cervical cancer screening.…”

Quantitative DNA methylation analysis of paired box gene 1 and LIM homeobox transcription factor 1 α genes in cervical cancer

“…In a previous study, LMX1A methylation testing demonstrated great potential for cervical lesion screening with a sensitivity, specificity and accuracy of 0.77, 0.88 and 0.90, respectively (30). In the present study, the pyrosequencing method was used to confirm the methylation of the LMX1A gene in cervical epithelial malignant transformation, but it was hardly methylated in LSIL.…”