Expression and physiological role of CCN4/Wnt-induced secreted protein 1 mRNA splicing variants in chondrocytes

FEBS J. 2007 Apr;274(7):1655-65. doi: 10.1111/j.1742-4658.2007.05709.x.

Abstract

CCN4/Wnt-induced secreted protein 1 (WISP1) is one of the CCN (CTGF/Cyr61/Nov) family proteins. CCN members have typical structures composed of four conserved cysteine-rich modules and their variants lacking certain modules, generated by alternative splicing or gene mutations, have been described in various pathological conditions. Several previous reports described a CCN4/WISP1 variant (WISP1v) lacking the second module in a few malignancies, but no information concerning the production of WISP1 variants in normal tissue is currently available. The expression of CCN4/WISP1 mRNA and its variants were analyzed in a human chondrosarcoma-derived chondrocytic cell line, HCS-2/8, and primary rabbit growth cartilage (RGC) chondrocytes. First, we found WISP1v and a novel variant of WISP1 (WISP1vx) to be expressed in HCS-2/8, as well as full-length WISP1 mRNA. This new variant was lacking the coding regions for the second and third modules and a small part of the first module. To monitor the expression of CCN4/WISP1 mRNA along chondrocyte differentiation, RGC cells were cultured and sampled until they were mineralized. As a result, we identified a WISP1v ortholog in normal RGC cells. Interestingly, the WISP1v mRNA level increased dramatically along with terminal differentiation. Furthermore, overexpression of WISP1v provoked expression of an alkaline phosphatase gene that is a marker of terminal differentiation in HCS-2/8 cells. These findings indicate that WISP1v thus plays a critical role in chondrocyte differentiation toward endochondral ossification, whereas HCS-2/8-specific WISP1vx may be associated with the transformed phenotypes of chondrosarcomas.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aggrecans / genetics
  • Alkaline Phosphatase / genetics
  • Alternative Splicing*
  • Animals
  • CCN Intercellular Signaling Proteins
  • Cartilage / metabolism
  • Cell Differentiation / genetics
  • Cell Line
  • Cell Line, Tumor
  • Cells, Cultured
  • Chondrocytes / metabolism*
  • Collagen Type II / genetics
  • DNA, Complementary / chemistry
  • DNA, Complementary / genetics
  • Embryo, Mammalian / metabolism
  • Gene Expression Profiling*
  • HeLa Cells
  • Humans
  • Intracellular Signaling Peptides and Proteins / genetics*
  • Intracellular Signaling Peptides and Proteins / physiology
  • Mice
  • Molecular Sequence Data
  • Protein Isoforms / genetics
  • Protein Isoforms / physiology
  • Proto-Oncogene Proteins / genetics*
  • Proto-Oncogene Proteins / physiology
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Rabbits
  • Sequence Analysis, RNA
  • Transfection

Substances

  • Aggrecans
  • CCN Intercellular Signaling Proteins
  • CCN4 protein, human
  • Collagen Type II
  • DNA, Complementary
  • Intracellular Signaling Peptides and Proteins
  • Protein Isoforms
  • Proto-Oncogene Proteins
  • RNA, Messenger
  • Alkaline Phosphatase

Associated data

  • GENBANK/EF025921
  • GENBANK/EF025922
  • GENBANK/EF025923