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.

Possible involvement of cyclooxygenase-2 in the recurrence of desmoid fibromatosis: case series and mini-review of the literature

“…DF is known to have a high incidence of local recurrence [2,13]. Recurrence is often problematic, as there are no clear treatments indicated for DF recurrence [17].…”

“…DF has also been reported to occur at sites of previous surgery or blunt trauma [2,11,12]. Sporadic cases show a mutation in the β-catenin gene on 3p21, resulting in overexpression of β-catenin in the nucleus [13,14]. 14β-catenin plays an important role in regulating the size of skin wounds and mediates the action of TGF-β, which is known to promote wound hyperplasia through proliferation of fibroblasts [13,14].…”

“…Approximately 80% of neoplasms show granular nuclear expression of β-catenin [13,14]. Histologic differential diagnosis is broad and encompasses a wide range of spindle cell lesions, ranging from scarring to abdominal gastrointestinal stromal tumors and liposarcoma with 'low grade' dedifferentiation [15,16].…”

“…Histologic differential diagnosis is broad and encompasses a wide range of spindle cell lesions, ranging from scarring to abdominal gastrointestinal stromal tumors and liposarcoma with 'low grade' dedifferentiation [15,16]. However, most neoplasms resembling fibroma have specific histological diagnostic characteristics and can be confidently diagnosed by the absence of nuclear β-catenin immunoreactivity [13,14].…”

“…Sporadic cases show a mutation in the β-catenin gene on 3p21, resulting in overexpression of β-catenin in the nucleus [13,14]. 14β-catenin plays an important role in regulating the size of skin wounds and mediates the action of TGF-β, which is known to promote wound hyperplasia through proliferation of fibroblasts [13,14]. Thus, it can be assumed that DF represents partially uncontrolled wound healing [2].…”