Phosphatidylinositol 3-kinase is involved in the protection of primary cultured human erythroid precursor cells from apoptosis

Yoshihito Haseyama, Ken Ichi Sawada, Atsushi Oda, Kazuki Koizumi, Hina Takano, Takashi Tarumi, Mitsufumi Nishio, Makoto Handa, Yasuo Ikeda, Takao Koike

Research output: Contribution to journalArticle

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Abstract

Little is known about the physiologic role of phosphatidylinositol 3- kinase (PI-3K) in the development of erythrocytes. Previous studies have shown that the effects of the PI-3K inhibitor wortmannin on erythropoietin (EPO)-dependent cell lines differed depending on the cell type used. Wortmannin inhibited EPO-induced differentiation of some cell lines without affecting their proliferation; however, the EPO-induced proliferation of other cell lines was inhibited by wortmannin. In neither case were signs of apoptosis observed. We have previously reported that signaling in highly purified human colony forming units-erythroid (CFU-E), generated in vitro from CD34+ cells, differed from that in EPO-dependent cell lines, in the current study, we examined the effects of a more specific PI-3K inhibitor (LY294002) on human CFU-E. We found that LY294002 dose-dependently inhibits the proliferation of erythroid progenitor cells with a half-maximal effect at 10 μmol/L LY294002. LY294002 at similar concentrations also induces apoptosis of these cells, as evidenced by the appearance of annexin V-binding cells and DNA fragmentation. The steady-state phosphorylation of AKT at Ser- 473 that occurs as a result of PI-3K activation was also inhibited by LY294002 at similar concentrations, suggesting that the effects of LY294002 are specific. Interestingly, the acceleration of apoptosis by LY294002 was observed in the presence or absence of EPO. Further, deprivation of EPO resulted in accelerated apoptosis irrespective of the presence of LY294002. Our study confirms and extends the finding that signaling in human primary cultured erythroid cells is significantly different from that in EPO- dependent cell lines. These data suggest that PI-3K has an antiapoptotic role in erythroid progenitor cells. In addition, 2 different pathways for the protection of primary erythroid cells from apoptosis likely exist: 1 independent of EPO that is LY294002-sensitive and one that is EPO-dependent and at least partly insensitive to LY294002.

Original languageEnglish
Pages (from-to)1568-1577
Number of pages10
JournalBlood
Volume94
Issue number5
Publication statusPublished - 1999 Sep 1
Externally publishedYes

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Phosphatidylinositol 3-Kinase
Erythroid Precursor Cells
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
Erythropoietin
Apoptosis
Cells
Cell Line
Erythroid Cells
Phosphorylation
Annexin A5
DNA Fragmentation
Cultured Cells
Erythrocytes
Chemical activation

ASJC Scopus subject areas

  • Hematology

Cite this

Haseyama, Y., Sawada, K. I., Oda, A., Koizumi, K., Takano, H., Tarumi, T., ... Koike, T. (1999). Phosphatidylinositol 3-kinase is involved in the protection of primary cultured human erythroid precursor cells from apoptosis. Blood, 94(5), 1568-1577.

Phosphatidylinositol 3-kinase is involved in the protection of primary cultured human erythroid precursor cells from apoptosis. / Haseyama, Yoshihito; Sawada, Ken Ichi; Oda, Atsushi; Koizumi, Kazuki; Takano, Hina; Tarumi, Takashi; Nishio, Mitsufumi; Handa, Makoto; Ikeda, Yasuo; Koike, Takao.

In: Blood, Vol. 94, No. 5, 01.09.1999, p. 1568-1577.

Research output: Contribution to journalArticle

Haseyama, Y, Sawada, KI, Oda, A, Koizumi, K, Takano, H, Tarumi, T, Nishio, M, Handa, M, Ikeda, Y & Koike, T 1999, 'Phosphatidylinositol 3-kinase is involved in the protection of primary cultured human erythroid precursor cells from apoptosis', Blood, vol. 94, no. 5, pp. 1568-1577.
Haseyama Y, Sawada KI, Oda A, Koizumi K, Takano H, Tarumi T et al. Phosphatidylinositol 3-kinase is involved in the protection of primary cultured human erythroid precursor cells from apoptosis. Blood. 1999 Sep 1;94(5):1568-1577.
Haseyama, Yoshihito ; Sawada, Ken Ichi ; Oda, Atsushi ; Koizumi, Kazuki ; Takano, Hina ; Tarumi, Takashi ; Nishio, Mitsufumi ; Handa, Makoto ; Ikeda, Yasuo ; Koike, Takao. / Phosphatidylinositol 3-kinase is involved in the protection of primary cultured human erythroid precursor cells from apoptosis. In: Blood. 1999 ; Vol. 94, No. 5. pp. 1568-1577.
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abstract = "Little is known about the physiologic role of phosphatidylinositol 3- kinase (PI-3K) in the development of erythrocytes. Previous studies have shown that the effects of the PI-3K inhibitor wortmannin on erythropoietin (EPO)-dependent cell lines differed depending on the cell type used. Wortmannin inhibited EPO-induced differentiation of some cell lines without affecting their proliferation; however, the EPO-induced proliferation of other cell lines was inhibited by wortmannin. In neither case were signs of apoptosis observed. We have previously reported that signaling in highly purified human colony forming units-erythroid (CFU-E), generated in vitro from CD34+ cells, differed from that in EPO-dependent cell lines, in the current study, we examined the effects of a more specific PI-3K inhibitor (LY294002) on human CFU-E. We found that LY294002 dose-dependently inhibits the proliferation of erythroid progenitor cells with a half-maximal effect at 10 μmol/L LY294002. LY294002 at similar concentrations also induces apoptosis of these cells, as evidenced by the appearance of annexin V-binding cells and DNA fragmentation. The steady-state phosphorylation of AKT at Ser- 473 that occurs as a result of PI-3K activation was also inhibited by LY294002 at similar concentrations, suggesting that the effects of LY294002 are specific. Interestingly, the acceleration of apoptosis by LY294002 was observed in the presence or absence of EPO. Further, deprivation of EPO resulted in accelerated apoptosis irrespective of the presence of LY294002. Our study confirms and extends the finding that signaling in human primary cultured erythroid cells is significantly different from that in EPO- dependent cell lines. These data suggest that PI-3K has an antiapoptotic role in erythroid progenitor cells. In addition, 2 different pathways for the protection of primary erythroid cells from apoptosis likely exist: 1 independent of EPO that is LY294002-sensitive and one that is EPO-dependent and at least partly insensitive to LY294002.",
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AU - Tarumi, Takashi

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