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.
(c) 2010 Wiley-Liss, Inc.
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