title: Innovative rapid gene methylation analysis of surgical margin tissues in head and neck cancer.
abstract: BACKGROUND
  Securing the negative surgical margin is the first step in surgical cancer treatment. However, tumor recurrence sometimes occurs even with histologically negative surgical margins. To detect minimal residual cancer cells in the deep margin intraoperatively, a time-efficient molecular approach is required.
  
  
  METHODS
  We established an innovative rapid quantitative methylation PCR (QMSP) assay, which consists of substantially time-minimized DNA extraction, bisulfite treatment, and QMSP assays. To demonstrate the feasibility of this procedure, 10 serial surgical specimens of primary head and neck squamous cell carcinoma (HNSCC) were evaluated by both rapid and conventional QMSP. Two frequently methylated genes in head and neck cancer, homeobox A9 (HOXA9) and endothelin receptor type B (EDNRB) were analyzed in 10 HNSCCs and surgical margin tissues, as well as normal muscle and oral mucosa samples.
  
  
  RESULTS
  The product quality of DNA extraction and bisulfite treatment using the time-saving procedure was comparable to the conventional procedure. In the QMSP assay, target gene methylation and reference gene methylation were equally detected by both the rapid and conventional method. Finally, relative results of rapid and conventional QMSP were quite similar to each other in tumors, margins, and normal tissues. The average total time required for the rapid QMSP procedure was less than 3 h and could be accomplished by a single person.
  
  
  CONCLUSIONS
  From the viewpoint of accuracy, cost, and time consumption, the innovative rapid QMSP maintains highly sensitive methylation detection accomplished within the time frame of a major ablative and reconstructive procedure.
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Masamichi
authors.initials: M
authors.lastname: Hayashi
authors.forename: Rafael
authors.initials: R
authors.lastname: Guerrero-Preston
authors.forename: Jun
authors.initials: J
authors.lastname: Okamura
authors.forename: Christina
authors.initials: C
authors.lastname: Michailidi
authors.forename: Zubair
authors.initials: Z
authors.lastname: Kahn
authors.forename: Xiufeng
authors.initials: X
authors.lastname: Li
authors.forename: Julie
authors.initials: J
authors.lastname: Ahn
authors.forename: Marla
authors.initials: M
authors.lastname: Goldsmith
authors.forename: Wayne
authors.initials: W
authors.lastname: Koch
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D001483:Base Sequence; D014408:Biomarkers, Tumor / Q000235:genetics*; D002294:Carcinoma, Squamous Cell / Q000175:diagnosis* / Q000235:genetics / Q000601:surgery; D019175:DNA Methylation; D004273:DNA, Neoplasm / Q000235:genetics; D005260:Female; D005500:Follow-Up Studies; D006258:Head and Neck Neoplasms / Q000175:diagnosis* / Q000235:genetics / Q000601:surgery; D018398:Homeodomain Proteins / Q000235:genetics; D006801:Humans; D007432:Intraoperative Period; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D009364:Neoplasm Recurrence, Local / Q000175:diagnosis* / Q000235:genetics / Q000601:surgery; D009367:Neoplasm Staging; D018365:Neoplasm, Residual / Q000175:diagnosis* / Q000235:genetics / Q000601:surgery; D016133:Polymerase Chain Reaction / Q000379:methods; D011379:Prognosis; D011401:Promoter Regions, Genetic / Q000235:genetics; D044023:Receptor, Endothelin B; D017466:Receptors, Endothelin / Q000235:genetics
chemical_list: D014408:Biomarkers, Tumor; D004273:DNA, Neoplasm; C000588596:EDNRB protein, human; D018398:Homeodomain Proteins; D044023:Receptor, Endothelin B; D017466:Receptors, Endothelin; C097899:homeobox protein HOXA9