title: α₁-Adrenoceptor stimulation enhances leukemia inhibitory factor-induced proliferation of mouse-induced pluripotent stem cells.
abstract: Since the clinical use of induced pluripotent stem (iPS) cells may overcome the current obstacles in stem cell-based therapy, the molecular mechanisms that regulate iPS cell proliferation are of great interest. Therefore, in the present study, we determined the involvement of α(1)-adrenoceptor in the proliferation of mouse iPS cells. The selective α(1)-adrenoceptor agonist l-phenylephrine dose-dependently increased the proliferation of mouse iPS cells cultured in a medium with leukemia inhibitory factor (LIF). Pretreatment with either selective α(1)-adrenoceptor antagonists or protein kinase C (PKC) inhibitors significantly inhibited l-phenylephrine-induced DNA synthesis. The treatment with an IP(3) receptor agonist significantly enhanced LIF-induced DNA synthesis. On the other hand, we confirmed that the intracellular calcium level was increased by the treatment with l-phenylephrine. Thus, intracellular calcium release or PKC activation induced by α(1)-adrenoceptor activation may lead to the enhancement of DNA synthesis. In addition, pretreatment with mitogen-activated protein kinase (MAPK) kinase (MEK) inhibitor PD98059 or phosphatidylinositol-3 phosphate kinase (PI3K) inhibitor LY294002 significantly inhibited l-phenylephrine-induced DNA synthesis. Treatment with l-phenylephrine significantly increased Akt or p44/42 MAPK phosphorylation. α(1)-Adrenoceptor expression in mouse iPS cells was confirmed by immunofluorescence staining and western blotting analysis. In mouse iPS cells cultured with LIF, stimulation with l-phenylephrine significantly increased the proportion of cells in the S and G(2)/M phases and decreased that in the G(1) phase. These results suggest that stimulation with α(1)-adrenoceptor may enhance DNA synthesis and proliferation of mouse iPS cells cultured with LIF via augmentation of both the MEK/MAPK and the PI3K/Akt pathways.
authors: Department of Pharmacology, National Defense Medical College, Tokorozawa, Saitama, Japan. tishizu@ndmc.ac.jp
authors: Toshiaki
authors: T
authors: Ishizuka
authors: Yasuhiro
authors: Y
authors: Watanabe
mesh_terms: D058646:Adrenergic alpha-1 Receptor Agonists / Q000494:pharmacology; D058668:Adrenergic alpha-1 Receptor Antagonists / Q000494:pharmacology; D000818:Animals; D018929:Cell Culture Techniques; D002453:Cell Cycle / Q000187:drug effects; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D004247:DNA / Q000096:biosynthesis; D004789:Enzyme Activation / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism; D053605:Leukemia Inhibitory Factor / Q000494:pharmacology*; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D010656:Phenylephrine / Q000494:pharmacology; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D011224:Prazosin / Q000494:pharmacology; D011493:Protein Kinase C / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism; D018340:Receptors, Adrenergic, alpha-1 / Q000378:metabolism*; D050791:STAT Transcription Factors / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
chemical_list: D058646:Adrenergic alpha-1 Receptor Agonists; D058668:Adrenergic alpha-1 Receptor Antagonists; D053605:Leukemia Inhibitory Factor; D016271:Proto-Oncogene Proteins c-myc; D018340:Receptors, Adrenergic, alpha-1; D050791:STAT Transcription Factors; D016159:Tumor Suppressor Protein p53; D010656:Phenylephrine; D004247:DNA; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D011493:Protein Kinase C; D020928:Mitogen-Activated Protein Kinases; D011224:Prazosin