title: Quantitative detection of TIMP-3 promoter hypermethylation and its prognostic significance in esophageal squamous cell carcinoma.
abstract: Tissue inhibitor of metalloproteinase-3 (TIMP-3) has an inhibitory effect on tumor development, growth and metastasis due to its interaction with matrix metalloproteinases (MMPs). We investigated the prognostic significance of TIMP-3 gene promoter methylation in esophageal squamous cell carcinoma (ESCC). TIMP-3 methylation was analyzed by MethyLight, a quantitative and methylation-specific PCR method. Hypermethylation was found in 9/51 (18%) surgically resected ESCC samples and was associated with reduced disease-free (p=0.0039) and overall survival (p=0.0047). Upon multivariate analysis, it was found to be an independent prognostic parameter for poor survival and was associated with earlier recurrence after surgery (p=0.0238), especially via pleural dissemination (p=0.001). Expression levels for TIMP-3 protein and for several MMPs were analyzed by Western blot analysis in 20 pairs of ESCC and adjacent normal tissue. The expression of MMP-2, -7 and -9 proteins in tumor tissue was significantly higher than in corresponding normal mucosa (p=0.0051, 0.0064 and 0.0004, respectively). In contrast, the expression of TIMP-3 protein in tumor tissue was significantly lower than in matched normal mucosa (p=0.0152). Tumors with TIMP-3 hypermethylation expressed lower levels of TIMP-3 protein compared to those without hypermethylation (p=0.0357). These results demonstrate that TIMP-3 hypermethylation is associated with lower TIMP-3 protein expression in ESCC and with poor patient survival due to a high frequency of recurrence by pleural dissemination.
authors.affiliation: Gastroenterologic Surgery, Department of Oncology, Division of Cancer Medicine, Graduate School of Medical Science, Kanazawa University, Ishikawa 920-8641, Japan. nino@surg2.m.kanazawa-u.ac.jp
authors.forename: Itasu
authors.initials: I
authors.lastname: Ninomiya
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Kawakami
authors.forename: Sachio
authors.initials: S
authors.lastname: Fushida
authors.forename: Takashi
authors.initials: T
authors.lastname: Fujimura
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Funaki
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Takamura
authors.forename: Hirohisa
authors.initials: H
authors.lastname: Kitagawa
authors.forename: Hisatoshi
authors.initials: H
authors.lastname: Nakagawara
authors.forename: Hidehiro
authors.initials: H
authors.lastname: Tajima
authors.forename: Masato
authors.initials: M
authors.lastname: Kayahara
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Ohta
mesh_terms: D002294:Carcinoma, Squamous Cell / Q000235:genetics / Q000378:metabolism*; D019175:DNA Methylation; D018572:Disease-Free Survival; D004938:Esophageal Neoplasms / Q000235:genetics / Q000378:metabolism*; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D009092:Mucous Membrane / Q000473:pathology; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D016133:Polymerase Chain Reaction; D011379:Prognosis; D016016:Proportional Hazards Models; D013447:Sulfites / Q000494:pharmacology; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000235:genetics* / Q000378:metabolism; D016896:Treatment Outcome
chemical_list: D013447:Sulfites; C088390:TIMP3 protein, human; D019717:Tissue Inhibitor of Metalloproteinase-3; C009279:sodium bisulfite