Increased D-type cyclin expression together with decreased cdc2 activity confers megakaryocytic differentiation of a human thrombopoietin-dependent hematopoietic cell line.
At the late phase of megakaryocytopoiesis, megakaryocytes undergo endomitosis, which is characterized by DNA replication without cell division. Although a number of cell cycle regulatory molecules have been identified, the precise roles of these molecules in megakaryocytic endomitosis are largely unknown. In a human interleukin-3-dependent cell line transfected with the thrombopoietin (TPO) receptor c-mpl (F-36P-mpl), either treatment with TPO or the overexpression of activated ras (Ha-Ras(G12V)) induced megakaryocytic maturation with polyploid formation. We found that TPO stimulation or Ha-Ras(G12V) expression led to up-regulation of cyclin D1, cyclin D2, and cyclin D3 expression. In addition, expression levels of cyclin A and cyclin B were reduced during the total course of both TPO- and Ha-Ras(G12V)-induced megakaryocytic differentiation, thereby leading to decreased cdc2 kinase activity. Neither the induced expression of cyclin D1, cyclin D2, or cyclin D3 nor the expression of a dominant negative form of cdc2 alone could induce megakaryocytic differentiation of F-36P-mpl cells. In contrast, overexpression of dominant negative cdc2 together with cyclin D1, cyclin D2, or cyclin D3 facilitated megakaryocytic differentiation in the absence of TPO. These results suggest that both D-type cyclin expression and decreased cdc2 kinase activity may participate in megakaryocytic differentiation.
Matsumura
I
I
Department of Hematology/Oncology, Osaka University Medical School, 2-2, Yamada-oka, Suita, Osaka 565-0871, Japan. matumura@bldon.med.osaka-u.ac.jp
Tanaka
H
H
Kawasaki
A
A
Odajima
J
J
Daino
H
H
Hashimoto
K
K
Wakao
H
H
Nakajima
K
K
Kato
T
T
Miyazaki
H
H
Kanakura
Y
Y
D016203:CDC2 Protein Kinase / Q000378:metabolism*; D002453:Cell Cycle / Q000187:drug effects; D002454:Cell Differentiation; D056741:Cyclin D; D019938:Cyclin D1 / Q000378:metabolism; D056742:Cyclin D2; D056743:Cyclin D3; D016213:Cyclins / Q000378:metabolism / Q000502:physiology*; D018076:DNA, Complementary / Q000378:metabolism; D005434:Flow Cytometry; D005799:Genes, Dominant; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D008533:Megakaryocytes / Q000166:cytology*; D010957:Plasmids; D016283:Proto-Oncogene Proteins p21(ras) / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D013926:Thrombopoietin / Q000378:metabolism* / Q000494:pharmacology; D013997:Time Factors; D014407:Tumor Cells, Cultured
C080336:CCND2 protein, human; C534696:CCND3 protein, human; D056741:Cyclin D; D056742:Cyclin D2; D056743:Cyclin D3; D016213:Cyclins; D018076:DNA, Complementary; D011994:Recombinant Proteins; D019938:Cyclin D1; D007544:Isopropyl Thiogalactoside; D013926:Thrombopoietin; D016203:CDC2 Protein Kinase; C501009:HRAS protein, human; D016283:Proto-Oncogene Proteins p21(ras)