Involvement of prolonged ras activation in thrombopoietin-induced megakaryocytic differentiation of a human factor-dependent hematopoietic cell line

Itaru Matsumura, Koichi Nakajima, Hiroshi Wakao, Seisuke Hattori, Koji Hashimoto, Hiroyuki Sugahara, Takashi Kato, Hiroshi Miyazaki, Toshio Hirano, Yuzuru Kanakura

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Abstract

Thrombopoietin (TPO) is a hematopoietic growth factor that plays fundamental roles is both megakaryopoiesis and thrombopoiesis through binding to its receptor, c-mpl. Although, TPO has been shown to activate various types of intracellular signaling molecules, such as the Janus family of protein tyrosine kinases, signal transducers and activators of transcription (STATs), and ras, the precise mechanisms underlying TPOinduced proliferation and differentiation remain unknown. In an effort to clarify the mechanisms of TPOinduced proliferation and differentiation, c-mpl was introduced into F- 36P, a human interleukin-3 (IL-3)dependent erythroleukemia cell line, and the effects of TPO on the c-mpl-transfected F-36P (F-36P-mpl) cells were investigated. F-36P-mpl cells were found to proliferate an differentiate at a high rate into mature megakaryocytes in response to TPO. Dominant-negative (dn) forms of STAT1, STAT3, STAT5, and ras were inducibly expressed in F- 36P-mpl cells, and their effects on TPQ-induced proliferation and megakaryocytic differentiation were analyzed. Among these dn molecules, both dn ras and STAT5 reduced TPO- or IL-3-induced proliferation of F-36P-mpl cells by ~30%, and only dn ras could inhibit TPO-induced megakaryocytic differentiation. In accord with this result, overexpression of activated ras (H-ras(G12C)) for 5 days led to megakaryocytic differentiation of F-36P-mpl cells. In a time course analysis on H-ras(G12v)-induced differentiation, activation of the ras pathway for 24 to 28 h was required and sufficient to induce megakaryocytic differentiation. Consistent with this result, the treatment of F-36P-mpl cells-with TPO was able to induce prolonged activation of ras for more than 24 h, whereas IL-3 had only a transient effect. These results suggest that prolonged ras activation may be involved in TPO-induced megakaryocytic differentiation.

Original languageEnglish
Pages (from-to)4282-4290
Number of pages9
JournalMolecular and Cellular Biology
Volume18
Issue number7
Publication statusPublished - 1998 Jul
Externally publishedYes

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Thrombopoietin
Cell Line
Interleukin-3
Thrombopoiesis
Leukemia, Erythroblastic, Acute
Megakaryocytes
Transducers
Protein-Tyrosine Kinases
Intercellular Signaling Peptides and Proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Matsumura, I., Nakajima, K., Wakao, H., Hattori, S., Hashimoto, K., Sugahara, H., ... Kanakura, Y. (1998). Involvement of prolonged ras activation in thrombopoietin-induced megakaryocytic differentiation of a human factor-dependent hematopoietic cell line. Molecular and Cellular Biology, 18(7), 4282-4290.

Involvement of prolonged ras activation in thrombopoietin-induced megakaryocytic differentiation of a human factor-dependent hematopoietic cell line. / Matsumura, Itaru; Nakajima, Koichi; Wakao, Hiroshi; Hattori, Seisuke; Hashimoto, Koji; Sugahara, Hiroyuki; Kato, Takashi; Miyazaki, Hiroshi; Hirano, Toshio; Kanakura, Yuzuru.

In: Molecular and Cellular Biology, Vol. 18, No. 7, 07.1998, p. 4282-4290.

Research output: Contribution to journalArticle

Matsumura, I, Nakajima, K, Wakao, H, Hattori, S, Hashimoto, K, Sugahara, H, Kato, T, Miyazaki, H, Hirano, T & Kanakura, Y 1998, 'Involvement of prolonged ras activation in thrombopoietin-induced megakaryocytic differentiation of a human factor-dependent hematopoietic cell line', Molecular and Cellular Biology, vol. 18, no. 7, pp. 4282-4290.
Matsumura, Itaru ; Nakajima, Koichi ; Wakao, Hiroshi ; Hattori, Seisuke ; Hashimoto, Koji ; Sugahara, Hiroyuki ; Kato, Takashi ; Miyazaki, Hiroshi ; Hirano, Toshio ; Kanakura, Yuzuru. / Involvement of prolonged ras activation in thrombopoietin-induced megakaryocytic differentiation of a human factor-dependent hematopoietic cell line. In: Molecular and Cellular Biology. 1998 ; Vol. 18, No. 7. pp. 4282-4290.
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abstract = "Thrombopoietin (TPO) is a hematopoietic growth factor that plays fundamental roles is both megakaryopoiesis and thrombopoiesis through binding to its receptor, c-mpl. Although, TPO has been shown to activate various types of intracellular signaling molecules, such as the Janus family of protein tyrosine kinases, signal transducers and activators of transcription (STATs), and ras, the precise mechanisms underlying TPOinduced proliferation and differentiation remain unknown. In an effort to clarify the mechanisms of TPOinduced proliferation and differentiation, c-mpl was introduced into F- 36P, a human interleukin-3 (IL-3)dependent erythroleukemia cell line, and the effects of TPO on the c-mpl-transfected F-36P (F-36P-mpl) cells were investigated. F-36P-mpl cells were found to proliferate an differentiate at a high rate into mature megakaryocytes in response to TPO. Dominant-negative (dn) forms of STAT1, STAT3, STAT5, and ras were inducibly expressed in F- 36P-mpl cells, and their effects on TPQ-induced proliferation and megakaryocytic differentiation were analyzed. Among these dn molecules, both dn ras and STAT5 reduced TPO- or IL-3-induced proliferation of F-36P-mpl cells by ~30{\%}, and only dn ras could inhibit TPO-induced megakaryocytic differentiation. In accord with this result, overexpression of activated ras (H-ras(G12C)) for 5 days led to megakaryocytic differentiation of F-36P-mpl cells. In a time course analysis on H-ras(G12v)-induced differentiation, activation of the ras pathway for 24 to 28 h was required and sufficient to induce megakaryocytic differentiation. Consistent with this result, the treatment of F-36P-mpl cells-with TPO was able to induce prolonged activation of ras for more than 24 h, whereas IL-3 had only a transient effect. These results suggest that prolonged ras activation may be involved in TPO-induced megakaryocytic differentiation.",
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AU - Wakao, Hiroshi

AU - Hattori, Seisuke

AU - Hashimoto, Koji

AU - Sugahara, Hiroyuki

AU - Kato, Takashi

AU - Miyazaki, Hiroshi

AU - Hirano, Toshio

AU - Kanakura, Yuzuru

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