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.
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.
Department of Veterinary Physiology, Biotherapy Human Resources Center (BK 21), College of Veterinary Medicine, Chonnam National University, Gwangju, Korea.
Jung Min
JM
Ryu
Min Young
MY
Lee
Seung Pil
SP
Yun
Ho Jae
HJ
Han
D020013:Calcium Signaling / Q000187:drug effects; D002453:Cell Cycle; D049109:Cell Proliferation; D002478:Cells, Cultured; D019938:Cyclin D1 / Q000378:metabolism*; D019927:Cyclin E / Q000378:metabolism*; D004261:DNA Replication; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005312:Fetal Blood / Q000166:cytology; D005947:Glucose / Q000378:metabolism*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000201:enzymology*; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010766:Phosphorylation; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt / Q000037:antagonists & inhibitors / Q000378:metabolism*; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D058570:TOR Serine-Threonine Kinases; D013997:Time Factors; D053773:Transforming Growth Factor beta1 / Q000235:genetics / Q000378:metabolism*; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
D019938:Cyclin D1; D019927:Cyclin E; D048049:Extracellular Signal-Regulated MAP Kinases; D005947:Glucose; D047908:Intracellular Signaling Peptides and Proteins; D020928:Mitogen-Activated Protein Kinases; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011493:Protein Kinase C; D047428:Protein Kinase Inhibitors; D017346:Protein Serine-Threonine Kinases; D051057:Proto-Oncogene Proteins c-akt; D012333:RNA, Messenger; D011994:Recombinant Proteins; D058570:TOR Serine-Threonine Kinases; D053773:Transforming Growth Factor beta1; D048051:p38 Mitogen-Activated Protein Kinases; C530104:CCND1 protein, human; C546842:MTOR protein, human