Delayed apoptosis and modulation of phospholipase D activity by plasmid containing mammalian cDNA in human neutrophils

Sun Young Lee, Ja Woong Kim, Jun O. Jin, Min Gyu Song, Joo In Park, Do Sik Min, Jong Young Kwak

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Phospholipase D (PLD) has been reported to have an anti-apoptotic role in neutrophils. This study examined the effects of plasmids containing the cDNA of PLD on the apoptosis of neutrophils. The apoptotic rate of neutrophils treated with the pCDNA3.1 plasmid was similar to that of the untreated cells after 24 h culture. However, the addition of pCDNA3.1 containing the cDNA of either human PLD1 (pCDNA3.1-PLD1) or -PLD2 (pCDNA3.1-PLD2) to the culture media with or without transfection reagent significantly decreased the rate of spontaneous apoptosis but not Fas-stimulated apoptosis and the decreased apoptosis was blocked by 1-butanol. pCDNA3.1-PLD blocked the cleavage of procaspase-3 and -8. The phorbol myristate acetate stimulated the PLD activities of pCDNA3.1-PLD-treated neutrophils but did not stimulate the activities of untreated or pCDNA3.1-treated neutrophils. The level of the PLD1 protein was higher in the cultured neutrophils with pCDNA3.1-PLD than with the media or pCDNA3.1. The spontaneous apoptosis of neutrophils was inhibited and the PLD1 expression level was increased by the linearized or promoterless forms of pCDNA3.1-PLD1 and the plasmids containing the cDNA of the enhanced green fluorescent protein (pEGFP) and EGFP-PLD1. These results suggest that the plasmids containing mammalian cDNA inhibit the spontaneous apoptosis of neutrophils and modulate PLD.

Original languageEnglish
Pages (from-to)1039-1047
Number of pages9
JournalBiochemical and Biophysical Research Communications
Volume347
Issue number4
DOIs
Publication statusPublished - 2006 Sep 8

Fingerprint

Phospholipase D
Neutrophils
Plasmids
Complementary DNA
Modulation
Apoptosis
1-Butanol
Tetradecanoylphorbol Acetate
Caspase 8
Caspase 3
Culture Media
Transfection

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Lee, Sun Young ; Kim, Ja Woong ; Jin, Jun O. ; Song, Min Gyu ; Park, Joo In ; Min, Do Sik ; Kwak, Jong Young. / Delayed apoptosis and modulation of phospholipase D activity by plasmid containing mammalian cDNA in human neutrophils. In: Biochemical and Biophysical Research Communications. 2006 ; Vol. 347, No. 4. pp. 1039-1047.
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Delayed apoptosis and modulation of phospholipase D activity by plasmid containing mammalian cDNA in human neutrophils. / Lee, Sun Young; Kim, Ja Woong; Jin, Jun O.; Song, Min Gyu; Park, Joo In; Min, Do Sik; Kwak, Jong Young.

In: Biochemical and Biophysical Research Communications, Vol. 347, No. 4, 08.09.2006, p. 1039-1047.

Research output: Contribution to journalArticle

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T1 - Delayed apoptosis and modulation of phospholipase D activity by plasmid containing mammalian cDNA in human neutrophils

AU - Lee, Sun Young

AU - Kim, Ja Woong

AU - Jin, Jun O.

AU - Song, Min Gyu

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AU - Min, Do Sik

AU - Kwak, Jong Young

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AB - Phospholipase D (PLD) has been reported to have an anti-apoptotic role in neutrophils. This study examined the effects of plasmids containing the cDNA of PLD on the apoptosis of neutrophils. The apoptotic rate of neutrophils treated with the pCDNA3.1 plasmid was similar to that of the untreated cells after 24 h culture. However, the addition of pCDNA3.1 containing the cDNA of either human PLD1 (pCDNA3.1-PLD1) or -PLD2 (pCDNA3.1-PLD2) to the culture media with or without transfection reagent significantly decreased the rate of spontaneous apoptosis but not Fas-stimulated apoptosis and the decreased apoptosis was blocked by 1-butanol. pCDNA3.1-PLD blocked the cleavage of procaspase-3 and -8. The phorbol myristate acetate stimulated the PLD activities of pCDNA3.1-PLD-treated neutrophils but did not stimulate the activities of untreated or pCDNA3.1-treated neutrophils. The level of the PLD1 protein was higher in the cultured neutrophils with pCDNA3.1-PLD than with the media or pCDNA3.1. The spontaneous apoptosis of neutrophils was inhibited and the PLD1 expression level was increased by the linearized or promoterless forms of pCDNA3.1-PLD1 and the plasmids containing the cDNA of the enhanced green fluorescent protein (pEGFP) and EGFP-PLD1. These results suggest that the plasmids containing mammalian cDNA inhibit the spontaneous apoptosis of neutrophils and modulate PLD.

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