Parenchyma-stromal interactions induce fibrosis by secreting CCN2 and promote osteoclastogenesis by stimulating RANKL and CD68 through activated TGF-β/BMP4 in ameloblastoma

J Oral Pathol Med. 2017 Jan;46(1):67-75. doi: 10.1111/jop.12467. Epub 2016 Jun 21.

Abstract

Background: Tumor parenchyma-stromal interactions affect the properties of tumors and their dynamics. Our group previously showed that secreted frizzled related protein (sFRP)-2 impairs bone formation and promotes bone invasion in ameloblastoma. However, the effects of the secreted growth factors CCN2, TGF-β, and BMP4 on stromal tissues in ameloblastoma remain unclear.

Materials and results: Thirty-five paraffin-embedded ameloblastoma cases, ameloblastoma-derived cell lines (AM-1), and primary cultures of ameloblastoma stromal fibroblasts (ASF) were used. Immunohistochemistry, MTT assay, Western blotting, and RT-PCR were performed on these samples. Parenchyma-stromal CCN2 overexpression correlated significantly with fibrous-type stroma, but not with myxoid-type stroma, suggesting a role of CCN2 in fibrosis (P < 0.05). Recombinant CCN2 induction of enhanced ASF proliferation in AM-1 medium supports this view. Conversely, BMP4 and TGF-β were expressed in myxoid-type fibroblasts, but little expression was found in parenchyma. RANKL-positive and CD68-positive stromal cell populations were significantly greater in myxoid-type tumor areas than in fibrous-type tumor areas, while a higher Ki-67 labeling index was recorded in ameloblastoma with fibrous-type stroma. These data suggest that stromal properties influence bone resorption-related activities and growth rates, respectively.

Conclusions: These results suggest that the effects of secreted growth factors are governed by ameloblastoma parenchyma-stromal interactions. CCN2 promotes fibrogenesis independent of TGF-β signaling. Absence of CCN2 expression is associated with a phenotypic switch to a myxoid-type microenvironment that is conducive for TGF-β/BMP4 signaling to promote osteoclastogenesis.

Keywords: ameloblastoma; ameloblastoma-derived cell lines cells; bone morphogenetic protein-4; connective tissue growth factor; transforming growth factor-beta.

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Ameloblastoma / metabolism*
  • Ameloblastoma / pathology
  • Antigens, CD / metabolism
  • Antigens, Differentiation, Myelomonocytic / metabolism
  • Bone Morphogenetic Protein 4 / metabolism
  • Bone Resorption / metabolism
  • Cell Proliferation
  • Connective Tissue Growth Factor / metabolism*
  • Female
  • Fibroblasts / metabolism
  • Fibrosis
  • Humans
  • Immunohistochemistry
  • Jaw Neoplasms / metabolism*
  • Jaw Neoplasms / pathology
  • Male
  • Middle Aged
  • Osteogenesis / physiology*
  • RANK Ligand / metabolism
  • Stromal Cells / metabolism
  • Transforming Growth Factor beta / metabolism
  • Tumor Cells, Cultured
  • Young Adult

Substances

  • Antigens, CD
  • Antigens, Differentiation, Myelomonocytic
  • BMP4 protein, human
  • Bone Morphogenetic Protein 4
  • CCN2 protein, human
  • CD68 antigen, human
  • RANK Ligand
  • Transforming Growth Factor beta
  • Connective Tissue Growth Factor