Granulocytic differentiation of HL-60 cells results in spontaneous apoptosis mediated by increased caspase expression

R. William; G. Watson; Ori D. Rotstein; Jean Parodo; Richard Bitar; David Hackam; John C. Marshall

doi:10.1016/S0014-5793(97)00779-5

Granulocytic differentiation of HL-60 cells results in spontaneous apoptosis mediated by increased caspase expression

R. William, G. Watson, Ori D. Rotstein, Jean Parodo, Richard Bitar, David Hackam, John C. Marshall

Research output: Contribution to journal › Article › peer-review

76 Scopus citations

Abstract

HL-60 cells differentiating into neutrophil-like cells die an apoptotic death in vitro. Susceptibility to apoptosis is associated with decreased Bcl-2 protein and mRNA expression; however, the effect of differentiation on the expression of proapoptotic caspases is unknown. Spontaneous apoptosis occurred 6 days after retinoic acid treatment. Western blotting showed loss of Bcl-2 by day 7, and new expression of ICE (caspase 1) and CPP32 (caspase 3) protein by day 2. Northern analysis demonstrated loss of Bcl-2 mRNA and increases in ICE mRNA by day 2; CPP32 mRNA was unchanged. Differential Bcl-2 and ICE mRNA expression was also found when granulocytic differentiation was stimulated by DMSO. Differentiated HL-60 cell lysates exhibited functional ICE proteolytic activity. De novo caspase expression was responsible for the development of spontaneous apoptosis, since specific inhibitors of ICE (YVAD-CMK) and CPP32 (DEVD-CHO), inhibited retinoic acid induced spontaneous apoptosis. Functional maturation and susceptibility to apoptosis are both inducible and linked in this granulocyte precursor cell line.

Original language	English (US)
Pages (from-to)	603-609
Number of pages	7
Journal	FEBS Letters
Volume	412
Issue number	3
DOIs	https://doi.org/10.1016/S0014-5793(97)00779-5
State	Published - Aug 4 1997
Externally published	Yes

Keywords

Apoptosis
Bcl-2
Caspase
HL-60
Retinoic acid

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

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10.1016/S0014-5793(97)00779-5

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title = "Granulocytic differentiation of HL-60 cells results in spontaneous apoptosis mediated by increased caspase expression",

abstract = "HL-60 cells differentiating into neutrophil-like cells die an apoptotic death in vitro. Susceptibility to apoptosis is associated with decreased Bcl-2 protein and mRNA expression; however, the effect of differentiation on the expression of proapoptotic caspases is unknown. Spontaneous apoptosis occurred 6 days after retinoic acid treatment. Western blotting showed loss of Bcl-2 by day 7, and new expression of ICE (caspase 1) and CPP32 (caspase 3) protein by day 2. Northern analysis demonstrated loss of Bcl-2 mRNA and increases in ICE mRNA by day 2; CPP32 mRNA was unchanged. Differential Bcl-2 and ICE mRNA expression was also found when granulocytic differentiation was stimulated by DMSO. Differentiated HL-60 cell lysates exhibited functional ICE proteolytic activity. De novo caspase expression was responsible for the development of spontaneous apoptosis, since specific inhibitors of ICE (YVAD-CMK) and CPP32 (DEVD-CHO), inhibited retinoic acid induced spontaneous apoptosis. Functional maturation and susceptibility to apoptosis are both inducible and linked in this granulocyte precursor cell line.",

keywords = "Apoptosis, Bcl-2, Caspase, HL-60, Retinoic acid",

author = "R. William and G. Watson and Rotstein, {Ori D.} and Jean Parodo and Richard Bitar and David Hackam and Marshall, {John C.}",

note = "Funding Information: The authors thank Immunex for the gift of Human ICE cDNA and Dr. V.H. Dixit for the CPP32 cDNA. The study was supported by grants from the Physicians Services Incorporated Foundation, the Surgical Intensive Care Unit, Toronto Hospital and the Medical Research Council of Canada.",

year = "1997",

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doi = "10.1016/S0014-5793(97)00779-5",

language = "English (US)",

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pages = "603--609",

journal = "FEBS Letters",

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T1 - Granulocytic differentiation of HL-60 cells results in spontaneous apoptosis mediated by increased caspase expression

AU - William, R.

AU - Watson, G.

AU - Rotstein, Ori D.

AU - Parodo, Jean

AU - Bitar, Richard

AU - Hackam, David

AU - Marshall, John C.

N1 - Funding Information: The authors thank Immunex for the gift of Human ICE cDNA and Dr. V.H. Dixit for the CPP32 cDNA. The study was supported by grants from the Physicians Services Incorporated Foundation, the Surgical Intensive Care Unit, Toronto Hospital and the Medical Research Council of Canada.

PY - 1997/8/4

Y1 - 1997/8/4

N2 - HL-60 cells differentiating into neutrophil-like cells die an apoptotic death in vitro. Susceptibility to apoptosis is associated with decreased Bcl-2 protein and mRNA expression; however, the effect of differentiation on the expression of proapoptotic caspases is unknown. Spontaneous apoptosis occurred 6 days after retinoic acid treatment. Western blotting showed loss of Bcl-2 by day 7, and new expression of ICE (caspase 1) and CPP32 (caspase 3) protein by day 2. Northern analysis demonstrated loss of Bcl-2 mRNA and increases in ICE mRNA by day 2; CPP32 mRNA was unchanged. Differential Bcl-2 and ICE mRNA expression was also found when granulocytic differentiation was stimulated by DMSO. Differentiated HL-60 cell lysates exhibited functional ICE proteolytic activity. De novo caspase expression was responsible for the development of spontaneous apoptosis, since specific inhibitors of ICE (YVAD-CMK) and CPP32 (DEVD-CHO), inhibited retinoic acid induced spontaneous apoptosis. Functional maturation and susceptibility to apoptosis are both inducible and linked in this granulocyte precursor cell line.

AB - HL-60 cells differentiating into neutrophil-like cells die an apoptotic death in vitro. Susceptibility to apoptosis is associated with decreased Bcl-2 protein and mRNA expression; however, the effect of differentiation on the expression of proapoptotic caspases is unknown. Spontaneous apoptosis occurred 6 days after retinoic acid treatment. Western blotting showed loss of Bcl-2 by day 7, and new expression of ICE (caspase 1) and CPP32 (caspase 3) protein by day 2. Northern analysis demonstrated loss of Bcl-2 mRNA and increases in ICE mRNA by day 2; CPP32 mRNA was unchanged. Differential Bcl-2 and ICE mRNA expression was also found when granulocytic differentiation was stimulated by DMSO. Differentiated HL-60 cell lysates exhibited functional ICE proteolytic activity. De novo caspase expression was responsible for the development of spontaneous apoptosis, since specific inhibitors of ICE (YVAD-CMK) and CPP32 (DEVD-CHO), inhibited retinoic acid induced spontaneous apoptosis. Functional maturation and susceptibility to apoptosis are both inducible and linked in this granulocyte precursor cell line.

KW - Apoptosis

KW - Bcl-2

KW - Caspase

KW - HL-60

KW - Retinoic acid

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Granulocytic differentiation of HL-60 cells results in spontaneous apoptosis mediated by increased caspase expression

Abstract

Keywords

ASJC Scopus subject areas

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