Endoglin expression regulates basal and TGF-beta1-induced extracellular matrix synthesis in cultured L6E9 myoblasts

Cell Physiol Biochem. 2004;14(4-6):301-10. doi: 10.1159/000080340.

Abstract

Background/aims: TGF-beta1 plays a major role in extracellular matrix (ECM) accumulation in tissue fibrosis. Connective tissue growth factor appears to play a critical role in this effect. Endoglin is a component of the transforming growth factor b (TGF-beta) receptor complex. Endoglin is upregulated by TGF-beta1, but its functional role in ECM regulation is unknown. Using rat myoblasts as a model system, we have assessed the role of endoglin on regulating CTGF expression and ECM synthesis and accumulation in the presence or absence of TGF-beta1.

Methods: L6E9 myoblast cell line was transfected with human endoglin, and collagen, fibronectin and CTGF production was assessed by Western blot and by proline incorporation to collagen proteins.

Results: Northern blot analysis revealed that parental rat myoblasts L6E9 do not express endogenous endoglin. Upon endoglin transfection, endoglin-expressing cells displayed a decreased CTGF expression and decreased collagen and fibronectin accumulation respect to mock transfectants. Northern blot analysis also revealed a decreased alpha2 (I) procollagen mRNA expression in endoglin transfectants. TGF-beta1 treatment induced an increase in CTGF expression and collagen synthesis and accumulation in L6E9 myoblasts. This effect was significantly lower in endoglin-transfected than in mock-transfected cells.

Conclusion: These results demonstrate that endoglin expression negatively regulates basal and TGF-beta1-induced CTGF and collagen expression and synthesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, CD
  • Cell Line
  • Collagen Type I / biosynthesis
  • Collagen Type I / genetics
  • Connective Tissue Growth Factor
  • Down-Regulation
  • Endoglin
  • Extracellular Matrix / genetics
  • Extracellular Matrix / metabolism*
  • Fibronectins / biosynthesis
  • Fibronectins / genetics
  • Humans
  • Immediate-Early Proteins / analysis
  • Immediate-Early Proteins / physiology
  • Intercellular Signaling Peptides and Proteins / analysis
  • Intercellular Signaling Peptides and Proteins / physiology
  • Myoblasts / chemistry
  • Myoblasts / drug effects
  • Myoblasts / metabolism*
  • RNA, Messenger / analysis
  • RNA, Messenger / metabolism
  • Rats
  • Receptors, Cell Surface
  • Transfection
  • Transforming Growth Factor beta / genetics
  • Transforming Growth Factor beta / pharmacology*
  • Transforming Growth Factor beta / physiology
  • Transforming Growth Factor beta1
  • Vascular Cell Adhesion Molecule-1 / biosynthesis
  • Vascular Cell Adhesion Molecule-1 / genetics
  • Vascular Cell Adhesion Molecule-1 / physiology*

Substances

  • Antigens, CD
  • CCN2 protein, human
  • CCN2 protein, rat
  • Collagen Type I
  • ENG protein, human
  • Endoglin
  • Fibronectins
  • Immediate-Early Proteins
  • Intercellular Signaling Peptides and Proteins
  • RNA, Messenger
  • Receptors, Cell Surface
  • TGFB1 protein, human
  • Tgfb1 protein, rat
  • Transforming Growth Factor beta
  • Transforming Growth Factor beta1
  • Vascular Cell Adhesion Molecule-1
  • Connective Tissue Growth Factor