Cell cycle- and protein kinase C-specific effects of resiniferatoxin and resiniferonol 9,13,14-ortho-phenylacetate in intestinal epithelial cells

Mark R. Frey, Jennifer A. Clark, Nicholas W. Bateman, Marcelo G. Kazanietz, Adrian R Black, Jennifer D Black

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

We have previously reported that protein kinase C (PKC) signaling can trigger hallmark events of cell cycle withdrawal in intestinal epithelial cells, including downregulation of cyclin D1, induction of p21Waf1/Cip1, and activation of the growth suppressor function of pocket proteins. In the current study, we compared the cell cycle- and PKC-specific effects of the vanilloid resiniferatoxin (RTX), its parent diterpene resiniferonol 9,13,14-ortho-phenylacetate (ROPA), and the PKC agonist PMA in the IEC-18 non-transformed intestinal crypt cell line. ROPA and PMA were found to produce strikingly similar alterations in cell cycle progression and PKC activity in IEC-18 cells, although PMA was approximately 1000-fold more potent in producing these effects. Both agents induced a transient PKC-dependent blockade in G 1→S progression associated with transient downregulation of cyclin D1 and induction of p21Waf1/Cip1. In contrast, RTX produced a prolonged PKC-independent cell cycle arrest in G0/G1 phase which was maintained for longer than 24 h. This arrest was vanilloid receptor-independent and associated with prolonged downregulation of cyclin D1 mRNA and protein, with little effect on levels of p21Waf1/Cip1. Combined exposure to RTX and ROPA produced a sustained and complete cell cycle blockade in IEC-18 cells, associated with depletion of cyclin D1 and sustained enhancement of p21Waf1/Cip1 levels. PMA, ROPA, RTX and the RTX/ROPA combination were capable of activating ERK1/2 signaling in IEC-18 cells, albeit with different kinetics. In contrast, only PMA and ROPA activated JNK1/2 and p38 in this system. Notably, some preparations of commercially obtained RTX produced effects indistinguishable from those of the RTX/ROPA combination, suggesting that certain batches of the compound may contain significant amounts of ROPA (or another PKC agonist activity). Together, these data demonstrate that structurally related compounds can produce similar cell cycle-specific effects but through distinct mechanisms. In addition, they add to a growing body of evidence that vanilloids can have antiproliferative effects in a variety of cell types.

Original languageEnglish (US)
Pages (from-to)1873-1886
Number of pages14
JournalBiochemical Pharmacology
Volume67
Issue number10
DOIs
StatePublished - May 15 2004

Fingerprint

Cell Cycle Proteins
Protein Kinase C
Epithelial Cells
Cyclin D1
Cells
Cell Cycle
Down-Regulation
TRPV Cation Channels
Cell Cycle Resting Phase
resiniferonol 9,13,14-ortho-phenylacetate
resiniferatoxin
Diterpenes
G1 Phase
Cell Cycle Checkpoints
Proteins
Chemical activation
Cell Line
Messenger RNA
Kinetics
Growth

Keywords

  • BIM I
  • Cell cycle
  • PDBu
  • PKC
  • PMA
  • Protein kinase C
  • ROPA
  • RTX
  • Resiniferatoxin
  • bisindolylmaleimide I
  • phorbol 12,13-dibutyrate
  • phorbol 12-myristate 13-acetate
  • protein kinase C
  • resiniferatoxin
  • resiniferonol 9,13,14-ortho-phenylacetate

ASJC Scopus subject areas

  • Biochemistry
  • Pharmacology

Cite this

Cell cycle- and protein kinase C-specific effects of resiniferatoxin and resiniferonol 9,13,14-ortho-phenylacetate in intestinal epithelial cells. / Frey, Mark R.; Clark, Jennifer A.; Bateman, Nicholas W.; Kazanietz, Marcelo G.; Black, Adrian R; Black, Jennifer D.

In: Biochemical Pharmacology, Vol. 67, No. 10, 15.05.2004, p. 1873-1886.

Research output: Contribution to journalArticle

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AU - Clark, Jennifer A.

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AU - Kazanietz, Marcelo G.

AU - Black, Adrian R

AU - Black, Jennifer D

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AB - We have previously reported that protein kinase C (PKC) signaling can trigger hallmark events of cell cycle withdrawal in intestinal epithelial cells, including downregulation of cyclin D1, induction of p21Waf1/Cip1, and activation of the growth suppressor function of pocket proteins. In the current study, we compared the cell cycle- and PKC-specific effects of the vanilloid resiniferatoxin (RTX), its parent diterpene resiniferonol 9,13,14-ortho-phenylacetate (ROPA), and the PKC agonist PMA in the IEC-18 non-transformed intestinal crypt cell line. ROPA and PMA were found to produce strikingly similar alterations in cell cycle progression and PKC activity in IEC-18 cells, although PMA was approximately 1000-fold more potent in producing these effects. Both agents induced a transient PKC-dependent blockade in G 1→S progression associated with transient downregulation of cyclin D1 and induction of p21Waf1/Cip1. In contrast, RTX produced a prolonged PKC-independent cell cycle arrest in G0/G1 phase which was maintained for longer than 24 h. This arrest was vanilloid receptor-independent and associated with prolonged downregulation of cyclin D1 mRNA and protein, with little effect on levels of p21Waf1/Cip1. Combined exposure to RTX and ROPA produced a sustained and complete cell cycle blockade in IEC-18 cells, associated with depletion of cyclin D1 and sustained enhancement of p21Waf1/Cip1 levels. PMA, ROPA, RTX and the RTX/ROPA combination were capable of activating ERK1/2 signaling in IEC-18 cells, albeit with different kinetics. In contrast, only PMA and ROPA activated JNK1/2 and p38 in this system. Notably, some preparations of commercially obtained RTX produced effects indistinguishable from those of the RTX/ROPA combination, suggesting that certain batches of the compound may contain significant amounts of ROPA (or another PKC agonist activity). Together, these data demonstrate that structurally related compounds can produce similar cell cycle-specific effects but through distinct mechanisms. In addition, they add to a growing body of evidence that vanilloids can have antiproliferative effects in a variety of cell types.

KW - BIM I

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KW - PKC

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KW - RTX

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KW - bisindolylmaleimide I

KW - phorbol 12,13-dibutyrate

KW - phorbol 12-myristate 13-acetate

KW - protein kinase C

KW - resiniferatoxin

KW - resiniferonol 9,13,14-ortho-phenylacetate

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