High glucose regulates cyclin D1/E of human mesenchymal stem cells through TGF-beta1 expression via Ca2+/PKC/MAPKs and PI3K/Akt/mTOR signal pathways

J Cell Physiol. 2010 Jul;224(1):59-70. doi: 10.1002/jcp.22091.

Abstract

The elucidation of factors that support human mesenchymal stem cells (hMSCs) growth has remained unresolved partly because of the reliance of many researchers on ill-defined, proprietary medium formulation. Thus, we investigated the effects of high glucose (D-glucose, 25 mM) on hMSCs proliferation. High glucose significantly increased [(3)H]-thymidine incorporation and cell-cycle regulatory protein expression levels compared with 5 mM D-glucose or 25 mM L-glucose. In addition, high glucose increased transforming growth factor-beta1 (TGF-beta(1)) mRNA and protein expression levels. High glucose-induced cell-cycle regulatory protein expression levels and [(3)H]-thymidine incorporation, which were inhibited by TGF-beta(1) siRNA transfection and TGF-beta(1) neutralizing antibody treatment. High glucose-induced phosphorylation of protein kinase C (PKC), p44/42 mitogen-activated protein kinases (MAPKs), p38 MAPK, Akt, and mammalian target of rapamycin (mTOR) in a time-dependent manner. Pretreatment of PKC inhibitors (staurosporine, 10(-6) M; bisindolylmaleimide I, 10(-6) M), LY 294002 (PI3 kinase inhibitor, 10(-6) M), Akt inhibitor (10(-5) M), PD 98059 (p44/42 MAPKs inhibitor, 10(-5) M), SB 203580 (p38 MAPK inhibitor, 10(-6) M), and rapamycin (mTOR inhibitor, 10(-8) M) blocked the high glucose-induced cellular proliferation and TGF-beta(1) protein expression. In conclusion, high glucose stimulated hMSCs proliferation through TGF-beta(1) expression via Ca(2+)/PKC/MAPKs as well as PI3K/Akt/mTOR signal pathways.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calcium Signaling* / drug effects
  • Cell Cycle
  • Cell Proliferation
  • Cells, Cultured
  • Cyclin D1 / metabolism*
  • Cyclin E / metabolism*
  • DNA Replication
  • Extracellular Signal-Regulated MAP Kinases / metabolism
  • Fetal Blood / cytology
  • Glucose / metabolism*
  • Humans
  • Intracellular Signaling Peptides and Proteins / antagonists & inhibitors
  • Intracellular Signaling Peptides and Proteins / metabolism*
  • Mesenchymal Stem Cells / drug effects
  • Mesenchymal Stem Cells / enzymology*
  • Mitogen-Activated Protein Kinases / antagonists & inhibitors
  • Mitogen-Activated Protein Kinases / metabolism*
  • Phosphatidylinositol 3-Kinases / metabolism*
  • Phosphoinositide-3 Kinase Inhibitors
  • Phosphorylation
  • Protein Kinase C / antagonists & inhibitors
  • Protein Kinase C / metabolism*
  • Protein Kinase Inhibitors / pharmacology
  • Protein Serine-Threonine Kinases / antagonists & inhibitors
  • Protein Serine-Threonine Kinases / metabolism*
  • Proto-Oncogene Proteins c-akt / antagonists & inhibitors
  • Proto-Oncogene Proteins c-akt / metabolism*
  • RNA Interference
  • RNA, Messenger / metabolism
  • Recombinant Proteins / metabolism
  • TOR Serine-Threonine Kinases
  • Time Factors
  • Transforming Growth Factor beta1 / genetics
  • Transforming Growth Factor beta1 / metabolism*
  • p38 Mitogen-Activated Protein Kinases / metabolism

Substances

  • CCND1 protein, human
  • Cyclin E
  • Intracellular Signaling Peptides and Proteins
  • Phosphoinositide-3 Kinase Inhibitors
  • Protein Kinase Inhibitors
  • RNA, Messenger
  • Recombinant Proteins
  • Transforming Growth Factor beta1
  • Cyclin D1
  • MTOR protein, human
  • Protein Serine-Threonine Kinases
  • Proto-Oncogene Proteins c-akt
  • TOR Serine-Threonine Kinases
  • Protein Kinase C
  • Extracellular Signal-Regulated MAP Kinases
  • Mitogen-Activated Protein Kinases
  • p38 Mitogen-Activated Protein Kinases
  • Glucose