Resveratrol induced serine phosphorylation of p53 causes apoptosis in a mutant p53 prostate cancer cell line

Hung Yun Lin, Ai Shih, Faith B. Davis, Heng Yuan Tang, Leon J. Martino, James A. Bennett, Paul J. Davis

Research output: Contribution to journalArticle

129 Citations (Scopus)

Abstract

Purpose: Resveratrol (Calbiochem, La Jolla, California) is a naturally occurring stilbene reported to cause apoptosis in various cultured cancer cells. In the current study the effect of resveratrol was determined in the androgen insensitive DU 145 prostate cancer cell line. Induction of apoptosis and activation of apoptosis related signal transduction pathways were measured. Materials and Methods: DU 145 cells were treated with resveratrol and apoptosis was measured by determining nucleosome content. Activation of mitogen activated protein kinase (MAPK) (extracellular signal-regulated kinase 1/2), p53 content and serine-15 phosphorylation of p53 were measured by immunoblot. Electrophoretic mobility shift assay of p53 binding to DNA, and measurement of p21 and glyceraldehyde-3-phosphate dehydrogenase messenger RNA were also done. Results: Resveratrol induced apoptosis in DU 145 cells. The stilbene activated MAPK and caused increased abundance of p53 and serine-15 phosphorylated p53. Resveratrol induced serine-15 phosphorylation of p53 was blocked by PD 98059 (Calbiochem), a MAPK kinase inhibitor, implicating MAPK activation in the phosphorylation of p53. PD 98059 also inhibited resveratrol induced apoptosis. These results suggest that apoptosis induction by resveratrol in DU 145 cells requires serine-15 phosphorylation of p53 by MAPK. Inhibition of MAPK dependent serine-15 phosphorylation resulted in reduced p53 binding to a p53 specific oligonucleotide on electrophoretic mobility shift assay. Pifithrin-α (Calbiochem), a p53 inhibitor, blocked resveratrol induced serine-15 phosphorylation of p53 and p53 binding to DNA. Resveratrol caused a p53 stimulated increase in p21 messenger RNA. Transfection of additional wild-type p53 into DU 145 cells induced apoptosis, which was further enhanced by resveratrol treatment. Conclusions: Resveratrol causes apoptosis in DU 145 prostate cancer cells. This action depends on the activation of MAPK, increase in cellular p53 content, serine-15 phosphorylation of p53 and increased p53 binding to DNA.

Original languageEnglish
Pages (from-to)748-755
Number of pages8
JournalJournal of Urology
Volume168
Issue number2
Publication statusPublished - 2002
Externally publishedYes

Fingerprint

Serine
Prostatic Neoplasms
Phosphorylation
Apoptosis
Cell Line
Mitogen-Activated Protein Kinases
Stilbenes
Electrophoretic Mobility Shift Assay
DNA
resveratrol
Messenger RNA
Glyceraldehyde-3-Phosphate Dehydrogenases
Mitogen-Activated Protein Kinase 3
Nucleosomes
Mitogen-Activated Protein Kinase 1
Mitogen-Activated Protein Kinase Kinases
Oligonucleotides
Androgens
Transfection
Cultured Cells

Keywords

  • Apoptosis
  • Gene expression
  • Genes, p53
  • Prostate
  • Prostatic neoplasms

ASJC Scopus subject areas

  • Urology

Cite this

Lin, H. Y., Shih, A., Davis, F. B., Tang, H. Y., Martino, L. J., Bennett, J. A., & Davis, P. J. (2002). Resveratrol induced serine phosphorylation of p53 causes apoptosis in a mutant p53 prostate cancer cell line. Journal of Urology, 168(2), 748-755.

Resveratrol induced serine phosphorylation of p53 causes apoptosis in a mutant p53 prostate cancer cell line. / Lin, Hung Yun; Shih, Ai; Davis, Faith B.; Tang, Heng Yuan; Martino, Leon J.; Bennett, James A.; Davis, Paul J.

In: Journal of Urology, Vol. 168, No. 2, 2002, p. 748-755.

Research output: Contribution to journalArticle

Lin, HY, Shih, A, Davis, FB, Tang, HY, Martino, LJ, Bennett, JA & Davis, PJ 2002, 'Resveratrol induced serine phosphorylation of p53 causes apoptosis in a mutant p53 prostate cancer cell line', Journal of Urology, vol. 168, no. 2, pp. 748-755.
Lin, Hung Yun ; Shih, Ai ; Davis, Faith B. ; Tang, Heng Yuan ; Martino, Leon J. ; Bennett, James A. ; Davis, Paul J. / Resveratrol induced serine phosphorylation of p53 causes apoptosis in a mutant p53 prostate cancer cell line. In: Journal of Urology. 2002 ; Vol. 168, No. 2. pp. 748-755.
@article{619ad0d88979498caf846987e04f8287,
title = "Resveratrol induced serine phosphorylation of p53 causes apoptosis in a mutant p53 prostate cancer cell line",
abstract = "Purpose: Resveratrol (Calbiochem, La Jolla, California) is a naturally occurring stilbene reported to cause apoptosis in various cultured cancer cells. In the current study the effect of resveratrol was determined in the androgen insensitive DU 145 prostate cancer cell line. Induction of apoptosis and activation of apoptosis related signal transduction pathways were measured. Materials and Methods: DU 145 cells were treated with resveratrol and apoptosis was measured by determining nucleosome content. Activation of mitogen activated protein kinase (MAPK) (extracellular signal-regulated kinase 1/2), p53 content and serine-15 phosphorylation of p53 were measured by immunoblot. Electrophoretic mobility shift assay of p53 binding to DNA, and measurement of p21 and glyceraldehyde-3-phosphate dehydrogenase messenger RNA were also done. Results: Resveratrol induced apoptosis in DU 145 cells. The stilbene activated MAPK and caused increased abundance of p53 and serine-15 phosphorylated p53. Resveratrol induced serine-15 phosphorylation of p53 was blocked by PD 98059 (Calbiochem), a MAPK kinase inhibitor, implicating MAPK activation in the phosphorylation of p53. PD 98059 also inhibited resveratrol induced apoptosis. These results suggest that apoptosis induction by resveratrol in DU 145 cells requires serine-15 phosphorylation of p53 by MAPK. Inhibition of MAPK dependent serine-15 phosphorylation resulted in reduced p53 binding to a p53 specific oligonucleotide on electrophoretic mobility shift assay. Pifithrin-α (Calbiochem), a p53 inhibitor, blocked resveratrol induced serine-15 phosphorylation of p53 and p53 binding to DNA. Resveratrol caused a p53 stimulated increase in p21 messenger RNA. Transfection of additional wild-type p53 into DU 145 cells induced apoptosis, which was further enhanced by resveratrol treatment. Conclusions: Resveratrol causes apoptosis in DU 145 prostate cancer cells. This action depends on the activation of MAPK, increase in cellular p53 content, serine-15 phosphorylation of p53 and increased p53 binding to DNA.",
keywords = "Apoptosis, Gene expression, Genes, p53, Prostate, Prostatic neoplasms",
author = "Lin, {Hung Yun} and Ai Shih and Davis, {Faith B.} and Tang, {Heng Yuan} and Martino, {Leon J.} and Bennett, {James A.} and Davis, {Paul J.}",
year = "2002",
language = "English",
volume = "168",
pages = "748--755",
journal = "Journal of Urology",
issn = "0022-5347",
publisher = "Elsevier Inc.",
number = "2",

}

TY - JOUR

T1 - Resveratrol induced serine phosphorylation of p53 causes apoptosis in a mutant p53 prostate cancer cell line

AU - Lin, Hung Yun

AU - Shih, Ai

AU - Davis, Faith B.

AU - Tang, Heng Yuan

AU - Martino, Leon J.

AU - Bennett, James A.

AU - Davis, Paul J.

PY - 2002

Y1 - 2002

N2 - Purpose: Resveratrol (Calbiochem, La Jolla, California) is a naturally occurring stilbene reported to cause apoptosis in various cultured cancer cells. In the current study the effect of resveratrol was determined in the androgen insensitive DU 145 prostate cancer cell line. Induction of apoptosis and activation of apoptosis related signal transduction pathways were measured. Materials and Methods: DU 145 cells were treated with resveratrol and apoptosis was measured by determining nucleosome content. Activation of mitogen activated protein kinase (MAPK) (extracellular signal-regulated kinase 1/2), p53 content and serine-15 phosphorylation of p53 were measured by immunoblot. Electrophoretic mobility shift assay of p53 binding to DNA, and measurement of p21 and glyceraldehyde-3-phosphate dehydrogenase messenger RNA were also done. Results: Resveratrol induced apoptosis in DU 145 cells. The stilbene activated MAPK and caused increased abundance of p53 and serine-15 phosphorylated p53. Resveratrol induced serine-15 phosphorylation of p53 was blocked by PD 98059 (Calbiochem), a MAPK kinase inhibitor, implicating MAPK activation in the phosphorylation of p53. PD 98059 also inhibited resveratrol induced apoptosis. These results suggest that apoptosis induction by resveratrol in DU 145 cells requires serine-15 phosphorylation of p53 by MAPK. Inhibition of MAPK dependent serine-15 phosphorylation resulted in reduced p53 binding to a p53 specific oligonucleotide on electrophoretic mobility shift assay. Pifithrin-α (Calbiochem), a p53 inhibitor, blocked resveratrol induced serine-15 phosphorylation of p53 and p53 binding to DNA. Resveratrol caused a p53 stimulated increase in p21 messenger RNA. Transfection of additional wild-type p53 into DU 145 cells induced apoptosis, which was further enhanced by resveratrol treatment. Conclusions: Resveratrol causes apoptosis in DU 145 prostate cancer cells. This action depends on the activation of MAPK, increase in cellular p53 content, serine-15 phosphorylation of p53 and increased p53 binding to DNA.

AB - Purpose: Resveratrol (Calbiochem, La Jolla, California) is a naturally occurring stilbene reported to cause apoptosis in various cultured cancer cells. In the current study the effect of resveratrol was determined in the androgen insensitive DU 145 prostate cancer cell line. Induction of apoptosis and activation of apoptosis related signal transduction pathways were measured. Materials and Methods: DU 145 cells were treated with resveratrol and apoptosis was measured by determining nucleosome content. Activation of mitogen activated protein kinase (MAPK) (extracellular signal-regulated kinase 1/2), p53 content and serine-15 phosphorylation of p53 were measured by immunoblot. Electrophoretic mobility shift assay of p53 binding to DNA, and measurement of p21 and glyceraldehyde-3-phosphate dehydrogenase messenger RNA were also done. Results: Resveratrol induced apoptosis in DU 145 cells. The stilbene activated MAPK and caused increased abundance of p53 and serine-15 phosphorylated p53. Resveratrol induced serine-15 phosphorylation of p53 was blocked by PD 98059 (Calbiochem), a MAPK kinase inhibitor, implicating MAPK activation in the phosphorylation of p53. PD 98059 also inhibited resveratrol induced apoptosis. These results suggest that apoptosis induction by resveratrol in DU 145 cells requires serine-15 phosphorylation of p53 by MAPK. Inhibition of MAPK dependent serine-15 phosphorylation resulted in reduced p53 binding to a p53 specific oligonucleotide on electrophoretic mobility shift assay. Pifithrin-α (Calbiochem), a p53 inhibitor, blocked resveratrol induced serine-15 phosphorylation of p53 and p53 binding to DNA. Resveratrol caused a p53 stimulated increase in p21 messenger RNA. Transfection of additional wild-type p53 into DU 145 cells induced apoptosis, which was further enhanced by resveratrol treatment. Conclusions: Resveratrol causes apoptosis in DU 145 prostate cancer cells. This action depends on the activation of MAPK, increase in cellular p53 content, serine-15 phosphorylation of p53 and increased p53 binding to DNA.

KW - Apoptosis

KW - Gene expression

KW - Genes, p53

KW - Prostate

KW - Prostatic neoplasms

UR - http://www.scopus.com/inward/record.url?scp=0036072056&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036072056&partnerID=8YFLogxK

M3 - Article

C2 - 12131363

AN - SCOPUS:0036072056

VL - 168

SP - 748

EP - 755

JO - Journal of Urology

JF - Journal of Urology

SN - 0022-5347

IS - 2

ER -