title: High-resolution melting analysis of HPV-16L1 gene methylation: A promising method for prognosing cervical cancer.
abstract: OBJECTIVE
  Methylation-sensitive high-resolution melting (MS-HRM) is a new technique for DNA methylation analysis, but it is rarely used for the detection of viral DNA methylation. In this study, we investigated the HPV-16L1 gene methylation that is detected by MS-HRM as a potential biomarker for prognosing cervical dysplasia and cancer.
  
  
  METHODS
  A total of 114 HPV-16 infected patients (normal (17), CIN1 (25), CIN2 (29), CIN3 (32), SCC (11)) who underwent liquid-based cytology test and biopsy were included in this study. 17 cases with HPV-16 infection and negative cytologic and histologic results served as the control group. The HPV-16L1 gene methylation statuses of these samples were investigated using a methylation-sensitive high-resolution melting (MS-HRM) assay after bisulfite modification.
  
  
  RESULTS
  The HPV-16L1 gene methylation statuses of all the 114 specimens were successfully detected by MS-HRM, and we observed increasing methylation levels in severe lesions, as determined using histologic assays. In addition, the methylation levels of CIN2+ (CIN2, CIN3 and SCC) were significantly higher than that of CIN2- (normal and CIN1, P<0.001). When taking CIN2+ as the reference, our HPV-16L1 DNA methylation assay achieved 91.7% sensitivity and 59.5% specificity, respectively.
  
  
  CONCLUSIONS
  The results of the present work demonstrated that HPV-16L1 gene methylation was closely associated with cervical precancerosis and cancer. Moreover, using MS-HRM to detect HPV-16L1 gene methylation may be a powerful assay for the triage of HPV-16-positive females, which could identify patients with high risk of invasive cancer.
authors.affiliation: Department of Cytopathology, The Third Affiliated Hospital of Zhengzhou University, No. 7 Front Kangfu Street, Er'qi District, Zhengzhou 450052, China.
authors.forename: Cui
authors.initials: C
authors.lastname: Qiu
authors.affiliation: Department of Cytopathology, The Third Affiliated Hospital of Zhengzhou University, No. 7 Front Kangfu Street, Er'qi District, Zhengzhou 450052, China.
authors.forename: Yanfang
authors.initials: Y
authors.lastname: Zhi
authors.affiliation: The Cancer Hospital Affiliated to Zhengzhou University, No. 127 Dongming Street, Jinshui District, Zhengzhou 450008, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Shen
authors.affiliation: The Second Affiliated Hospital of Zhengzhou University, No. 2 Jingba Street, Guancheng Hui District, Zhengzhou 450014, China.
authors.forename: Jiaomei
authors.initials: J
authors.lastname: Gong
authors.affiliation: Department of Cytopathology, The Third Affiliated Hospital of Zhengzhou University, No. 7 Front Kangfu Street, Er'qi District, Zhengzhou 450052, China.
authors.forename: Ya
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Cytopathology, The Third Affiliated Hospital of Zhengzhou University, No. 7 Front Kangfu Street, Er'qi District, Zhengzhou 450052, China. Electronic address: qc1213@outlook.com.
authors.forename: Xiaofu
authors.initials: X
authors.lastname: Li
mesh_terms: D000328:Adult; D000368:Aged; D036022:Capsid Proteins / Q000235:genetics; D019175:DNA Methylation / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics; D005260:Female; D005453:Fluorescence; D006801:Humans; D016014:Linear Models; D008875:Middle Aged; D060787:Neoplasm Grading; D009691:Nucleic Acid Denaturation / Q000235:genetics*; D009856:Oncogene Proteins, Viral / Q000235:genetics; D011379:Prognosis; D012015:Reference Standards; D002583:Uterine Cervical Neoplasms / Q000175:diagnosis* / Q000235:genetics* / Q000473:pathology
chemical_list: D036022:Capsid Proteins; D004279:DNA, Viral; D009856:Oncogene Proteins, Viral; C063317:L1 protein, Human papillomavirus type 16
keywords: Cervical cancer; Cervical intraepithelial neoplasia; DNA methylation; HPV-16; High-resolution melting; L1 gene