title: Characteristics of cultured desmoid cells with different CTNNB1 mutation status.
abstract: Desmoid tumors are benign mesenchymal neoplasms with a locally aggressive nature. The mutational status of β-catenin gene (CTNNB1) is presumed to affect the tumorous activity of the cells. In this study, we isolated three kinds of desmoid cell with different CTNNB1 status, and compared their characteristics. Cells were isolated from three patients with abdominal wall desmoid during surgery, all of which were resistant to meloxicam treatment. The mutational status of the CTNNB1 exon 3 was determined for both parental tumor tissues and isolated cultured cells. β-catenin expression was determined with immunohistochemistry. Responsiveness to meloxicam was investigated with MTS assay together with COX-2 immunostaining. mRNA expressions of downstream molecules of Wnt/β-catenin pathway were determined with real-time RT-PCR. Three kinds of cell isolated from desmoid tumors harboring different CTNNB1 mutation status (wild type, T41A, and S45F), all exhibited a spindle shape. These isolated cells could be cultured until the 20th passage with unchanged proliferative activity. Nuclear accumulation of β-catenin was observed in all cultured cells, particularly in those with S45F. Proliferating activity was significantly suppressed by meloxicam (25 μmol/L, P < 0.007) in all three cell cultures, of which parental desmoid was resistant to meloxicam clinically. The mRNA expressions of Axin2, c-Myc, and Cyclin D1 differently increased in the three cultured cell types as compared with those in human skin fibroblast cells (HDF). Inhibitors of Wnt/β-catenin pathway downregulated Axin2, c-Myc, and Cyclin D1 significantly. Isolated and cultured desmoid tumor cells harboring any one of the CTNNB1 mutation status had unique characteristics, and could be useful to investigate desmoid tumors with different mutation status of CTNNB1.
authors.affiliation: Department of Orthopaedic Surgery, Nagoya University Graduate School and School of Medicine, Nagoya, Japan.
authors.forename: Shunsuke
authors.initials: S
authors.lastname: Hamada
authors.affiliation: Department of Orthopaedic Surgery, Nagoya University Graduate School and School of Medicine, Nagoya, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Urakawa
authors.affiliation: Department of Orthopaedic Surgery, Nagoya University Graduate School and School of Medicine, Nagoya, Japan.
authors.forename: Eiji
authors.initials: E
authors.lastname: Kozawa
authors.affiliation: Department of Orthopaedic Surgery, Nagoya University Graduate School and School of Medicine, Nagoya, Japan.
authors.forename: Eisuke
authors.initials: E
authors.lastname: Arai
authors.affiliation: Department of Orthopaedic Surgery, Nagoya University Graduate School and School of Medicine, Nagoya, Japan.
authors.forename: Kunihiro
authors.initials: K
authors.lastname: Ikuta
authors.affiliation: Department of Orthopaedic Surgery, Nagoya University Graduate School and School of Medicine, Nagoya, Japan.
authors.forename: Tomohisa
authors.initials: T
authors.lastname: Sakai
authors.affiliation: Department of Orthopaedic Surgery, Nagoya University Graduate School and School of Medicine, Nagoya, Japan.
authors.forename: Naoki
authors.initials: N
authors.lastname: Ishiguro
authors.affiliation: Department of Orthopaedic Surgery, Nagoya University Graduate School and School of Medicine, Nagoya, Japan.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Nishida
mesh_terms: D000328:Adult; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D060466:Axin Protein / Q000235:genetics; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D019938:Cyclin D1 / Q000235:genetics; D051546:Cyclooxygenase 2 / Q000235:genetics / Q000378:metabolism; D004252:DNA Mutational Analysis; D018222:Desmoid Tumors / Q000235:genetics*; D015870:Gene Expression; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D016259:Genes, myc; D006801:Humans; D000077239:Meloxicam; D009154:Mutation; D012333:RNA, Messenger / Q000235:genetics; D013843:Thiazines / Q000494:pharmacology; D013844:Thiazoles / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D060449:Wnt Signaling Pathway / Q000187:drug effects; D055815:Young Adult; D051176:beta Catenin / Q000235:genetics*
chemical_list: C491761:AXIN2 protein, human; D000970:Antineoplastic Agents; D060466:Axin Protein; C495270:CTNNB1 protein, human; D012333:RNA, Messenger; D013843:Thiazines; D013844:Thiazoles; D051176:beta Catenin; D019938:Cyclin D1; D051546:Cyclooxygenase 2; D000077239:Meloxicam
keywords: CTNNB1 mutation; Wnt/β-catenin pathway; desmoid tumor; meloxicam; β-Catenin