title: PI3K/Akt/FOXO3a pathway contributes to thrombopoietin-induced proliferation of primary megakaryocytes in vitro and in vivo via modulation of p27(Kip1).
abstract: Thrombopoietin (TPO), the primary regulator of megakaryocyte (MK) and platelet formation, modulates the activity of multiple signal transduction molecules, including those in the Jak/STAT, p42/p44 MAPK and phosphatidylinositol 3-kinase (PI3K)/Akt pathways. We previously demonstrated that PI3K and Akt are necessary for TPO-induced cell cycle progression of primary MK progenitors. However, the molecular events secondary to the activation of PI3K/Akt responsible for MK proliferation remain unclear. In this study we show that FOXO3a and its downstream target p27(Kip1) play an important role in TPO-induced proliferation of MK progenitors. We found that TPO down-modulates p27(Kip1) expression at both the mRNA and protein levels in primary MKs in a PI3K dependent fashion. UT-7/TPO, a megakaryocytic cell line, stably expressing constitutively active Akt or a dominant-negative form of FOXO3a failed to reduce p27(Kip1) expression after TPO stimulation, and fail to induce p27(Kip1) expression following TPO withdrawal. Induced expression of an active form of FOXO3a resulted in increased p27(Kip1) expression in this cell line. In addition, the number of MKs is significantly increased in bone marrow from Foxo3a-deficient mice. Taken together with the previous observation that p27(Kip1)-deficient mice also display increased numbers of MK progenitors, our findings indicate that the PI3K/Akt/FOXO3a/p27(Kip1) pathway contributes to normal TPO-induced MK proliferation.
authors.affiliation: Department of Medicine, Division of Hematology/Oncology, University of California San Diego, La Jolla, California, USA. nakao-t@med.osaka-cu.ac.jp
authors.forename: Takafumi
authors.initials: T
authors.lastname: Nakao
authors.forename: Amy E
authors.initials: AE
authors.lastname: Geddis
authors.forename: Norma E
authors.initials: NE
authors.lastname: Fox
authors.forename: Kenneth
authors.initials: K
authors.lastname: Kaushansky
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002460:Cell Line; D049109:Cell Proliferation; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000378:metabolism*; D000071316:Forkhead Box Protein O3; D051858:Forkhead Transcription Factors / Q000235:genetics / Q000378:metabolism*; D008533:Megakaryocytes / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D015398:Signal Transduction; D013926:Thrombopoietin / Q000378:metabolism*
chemical_list: C493961:Cdkn1b protein, mouse; D000071316:Forkhead Box Protein O3; D051858:Forkhead Transcription Factors; C474033:FoxO3 protein, mouse; D050760:Cyclin-Dependent Kinase Inhibitor p27; D013926:Thrombopoietin; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt