Identification of DNA methylation of SOX9 in cervical cancer using methylated-CpG             island recovery assay.
The aim of the present study was to identify novel methylation markers for             cervical cancer screening and to test the clinical application of the most promising             biomarker in cervical scrapings. Methylated-CpG island recovery assay-based microarray             analysis was carried out on a discovery set consisting of cervical cancer tissue             and normal cervical tissue to identify significantly hypermethylated genes. Five             hundred and four CpG islands, corresponding to 378 genes, were differentially             methylated between cervical cancer tissue and normal cervical tissue. Among them,             30 genes were significantly hypermethylated. Of the 30 genes, SOX9, PKLR and DLX4             were selected for further validation by direct bisulfite sequencing. The SOX9             gene revealed complete methylation in the cervical cancer tissue and complete             non-methylation in the normal control tissue. A TaqMan-based real-time PCR assay             was performed to detect the methylation levels of the SOX9 gene in 156 cervical             scrapings, including 48 normal cervical scrapings, 30 scrapings with cervical             intraepithelial neoplasia 1 (CIN1), 30 scrapings with CIN2-3 and 48 scrapings             with squamous cell carcinoma (SCC). The methylation levels (methylation score)             of the SOX9 gene increased significantly with the severity of cervical squamous             lesions. The area under the receiver operating characteristic (ROC) curve (AUC)             revealed that the methylation score of the SOX9 gene could be used to segregate             SCC/CIN2-3 from CIN1/normal (AUC, 0.961; p=0.000). At the optimal cut-off value,             a sensitivity of 92.3% and a specificity of 89.7% were obtained. In conclusion,             SOX9 methylation is frequently involved in cervical carcinogenesis, and may provide             a valuable molecular biomarker for early detection of cervical cancer.
Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing 100026, PR China.
Jian-Hong
JH
Wu
Xue-Ai
XA
Liang
Yu-Mei
YM
Wu
Feng-Shuang
FS
Li
Yin-Mei
YM
Dai
D000328:Adult; D014408:Biomarkers, Tumor / Q000235:genetics*; D002294:Carcinoma, Squamous Cell / Q000235:genetics / Q000473:pathology; D016022:Case-Control Studies; D002584:Cervix Uteri / Q000378:metabolism* / Q000473:pathology; D018899:CpG Islands / Q000235:genetics*; D019175:DNA Methylation; D055088:Early Detection of Cancer; D005260:Female; D005500:Follow-Up Studies; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D011379:Prognosis; D012372:ROC Curve; D060888:Real-Time Polymerase Chain Reaction; D055758:SOX9 Transcription Factor / Q000235:genetics*; D017422:Sequence Analysis, DNA; D002583:Uterine Cervical Neoplasms / Q000235:genetics* / Q000473:pathology; D002578:Uterine Cervical Dysplasia / Q000235:genetics / Q000473:pathology
D014408:Biomarkers, Tumor; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human