Mitochondria as the primary target of resveratrol-induced apoptosis in human retinoblastoma cells

Dhruv Sareen, Paul R. Van Ginkel, Jennifer C. Takach, Ayesha Mohiuddin, Soesiawati R. Darjatmoko, Daniel Albert, Arthur S. Polans

Research output: Contribution to journalArticle

73 Citations (Scopus)

Abstract

PURPOSE. To determine the molecular mechanisms by which resveratrol induces retinoblastoma tumor cell death. METHODS. After resveratrol treatment, Y79 tumor cell viability was measured using a fluorescence-based assay, and proapoptotic and antiproliferative effects were characterized by Hoechst stain and flow cytometry, respectively. Mitochondrial transmembrane potential (ΔΨm) was measured as a function of drug treatment using 5,5′,6,6′-tetrachloro-1,1′,3,3′- tetraethylbenzamidazolocarbocyanin iodide (JC-1), whereas the release of cytochrome c from mitochondria was assayed by immunoblotting and caspase activities were determined by monitoring the cleavage of fluorogenic peptide substrates. RESULTS. Resveratrol induced a dose- and time-dependent decrease in Y79 tumor cell viability and inhibited proliferation by inducing S-phase growth arrest and apoptotic cell death. Preceding cell death, resveratrol evoked a rapid dissipation of ΔΨm. This was followed by the release of cytochrome c into the cytoplasm and a substantial increase in the activities of caspase-9 and caspase-3. Additionally, in a cell-free system, resveratrol directly induced the depolarization of isolated mitochondria. CONCLUSIONS. These results demonstrate that resveratrol, a nontoxic natural plant compound, inhibits Y79 cell proliferation and stimulates apoptosis through activation of the mitochondrial (intrinsic) apoptotic pathway and may warrant further exploration as an adjuvant to conventional anticancer therapies for retinoblastoma.

Original languageEnglish (US)
Pages (from-to)3708-3716
Number of pages9
JournalInvestigative Ophthalmology and Visual Science
Volume47
Issue number9
DOIs
StatePublished - Sep 1 2006
Externally publishedYes

Fingerprint

Retinoblastoma
Mitochondria
Apoptosis
Cell Death
Cytochromes c
Cell Survival
Neoplasms
Cell-Free System
Caspase 9
Iodides
Caspases
S Phase
Fluorescent Dyes
Immunoblotting
Caspase 3
Membrane Potentials
resveratrol
Flow Cytometry
Cytoplasm
Coloring Agents

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Mitochondria as the primary target of resveratrol-induced apoptosis in human retinoblastoma cells. / Sareen, Dhruv; Van Ginkel, Paul R.; Takach, Jennifer C.; Mohiuddin, Ayesha; Darjatmoko, Soesiawati R.; Albert, Daniel; Polans, Arthur S.

In: Investigative Ophthalmology and Visual Science, Vol. 47, No. 9, 01.09.2006, p. 3708-3716.

Research output: Contribution to journalArticle

Sareen, Dhruv ; Van Ginkel, Paul R. ; Takach, Jennifer C. ; Mohiuddin, Ayesha ; Darjatmoko, Soesiawati R. ; Albert, Daniel ; Polans, Arthur S. / Mitochondria as the primary target of resveratrol-induced apoptosis in human retinoblastoma cells. In: Investigative Ophthalmology and Visual Science. 2006 ; Vol. 47, No. 9. pp. 3708-3716.
@article{3a338f6f13e84c05930495a7063da5d9,
title = "Mitochondria as the primary target of resveratrol-induced apoptosis in human retinoblastoma cells",
abstract = "PURPOSE. To determine the molecular mechanisms by which resveratrol induces retinoblastoma tumor cell death. METHODS. After resveratrol treatment, Y79 tumor cell viability was measured using a fluorescence-based assay, and proapoptotic and antiproliferative effects were characterized by Hoechst stain and flow cytometry, respectively. Mitochondrial transmembrane potential (ΔΨm) was measured as a function of drug treatment using 5,5′,6,6′-tetrachloro-1,1′,3,3′- tetraethylbenzamidazolocarbocyanin iodide (JC-1), whereas the release of cytochrome c from mitochondria was assayed by immunoblotting and caspase activities were determined by monitoring the cleavage of fluorogenic peptide substrates. RESULTS. Resveratrol induced a dose- and time-dependent decrease in Y79 tumor cell viability and inhibited proliferation by inducing S-phase growth arrest and apoptotic cell death. Preceding cell death, resveratrol evoked a rapid dissipation of ΔΨm. This was followed by the release of cytochrome c into the cytoplasm and a substantial increase in the activities of caspase-9 and caspase-3. Additionally, in a cell-free system, resveratrol directly induced the depolarization of isolated mitochondria. CONCLUSIONS. These results demonstrate that resveratrol, a nontoxic natural plant compound, inhibits Y79 cell proliferation and stimulates apoptosis through activation of the mitochondrial (intrinsic) apoptotic pathway and may warrant further exploration as an adjuvant to conventional anticancer therapies for retinoblastoma.",
author = "Dhruv Sareen and {Van Ginkel}, {Paul R.} and Takach, {Jennifer C.} and Ayesha Mohiuddin and Darjatmoko, {Soesiawati R.} and Daniel Albert and Polans, {Arthur S.}",
year = "2006",
month = "9",
day = "1",
doi = "10.1167/iovs.06-0119",
language = "English (US)",
volume = "47",
pages = "3708--3716",
journal = "Investigative Ophthalmology and Visual Science",
issn = "0146-0404",
publisher = "Association for Research in Vision and Ophthalmology Inc.",
number = "9",

}

TY - JOUR

T1 - Mitochondria as the primary target of resveratrol-induced apoptosis in human retinoblastoma cells

AU - Sareen, Dhruv

AU - Van Ginkel, Paul R.

AU - Takach, Jennifer C.

AU - Mohiuddin, Ayesha

AU - Darjatmoko, Soesiawati R.

AU - Albert, Daniel

AU - Polans, Arthur S.

PY - 2006/9/1

Y1 - 2006/9/1

N2 - PURPOSE. To determine the molecular mechanisms by which resveratrol induces retinoblastoma tumor cell death. METHODS. After resveratrol treatment, Y79 tumor cell viability was measured using a fluorescence-based assay, and proapoptotic and antiproliferative effects were characterized by Hoechst stain and flow cytometry, respectively. Mitochondrial transmembrane potential (ΔΨm) was measured as a function of drug treatment using 5,5′,6,6′-tetrachloro-1,1′,3,3′- tetraethylbenzamidazolocarbocyanin iodide (JC-1), whereas the release of cytochrome c from mitochondria was assayed by immunoblotting and caspase activities were determined by monitoring the cleavage of fluorogenic peptide substrates. RESULTS. Resveratrol induced a dose- and time-dependent decrease in Y79 tumor cell viability and inhibited proliferation by inducing S-phase growth arrest and apoptotic cell death. Preceding cell death, resveratrol evoked a rapid dissipation of ΔΨm. This was followed by the release of cytochrome c into the cytoplasm and a substantial increase in the activities of caspase-9 and caspase-3. Additionally, in a cell-free system, resveratrol directly induced the depolarization of isolated mitochondria. CONCLUSIONS. These results demonstrate that resveratrol, a nontoxic natural plant compound, inhibits Y79 cell proliferation and stimulates apoptosis through activation of the mitochondrial (intrinsic) apoptotic pathway and may warrant further exploration as an adjuvant to conventional anticancer therapies for retinoblastoma.

AB - PURPOSE. To determine the molecular mechanisms by which resveratrol induces retinoblastoma tumor cell death. METHODS. After resveratrol treatment, Y79 tumor cell viability was measured using a fluorescence-based assay, and proapoptotic and antiproliferative effects were characterized by Hoechst stain and flow cytometry, respectively. Mitochondrial transmembrane potential (ΔΨm) was measured as a function of drug treatment using 5,5′,6,6′-tetrachloro-1,1′,3,3′- tetraethylbenzamidazolocarbocyanin iodide (JC-1), whereas the release of cytochrome c from mitochondria was assayed by immunoblotting and caspase activities were determined by monitoring the cleavage of fluorogenic peptide substrates. RESULTS. Resveratrol induced a dose- and time-dependent decrease in Y79 tumor cell viability and inhibited proliferation by inducing S-phase growth arrest and apoptotic cell death. Preceding cell death, resveratrol evoked a rapid dissipation of ΔΨm. This was followed by the release of cytochrome c into the cytoplasm and a substantial increase in the activities of caspase-9 and caspase-3. Additionally, in a cell-free system, resveratrol directly induced the depolarization of isolated mitochondria. CONCLUSIONS. These results demonstrate that resveratrol, a nontoxic natural plant compound, inhibits Y79 cell proliferation and stimulates apoptosis through activation of the mitochondrial (intrinsic) apoptotic pathway and may warrant further exploration as an adjuvant to conventional anticancer therapies for retinoblastoma.

UR - http://www.scopus.com/inward/record.url?scp=33749128952&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33749128952&partnerID=8YFLogxK

U2 - 10.1167/iovs.06-0119

DO - 10.1167/iovs.06-0119

M3 - Article

C2 - 16936077

AN - SCOPUS:33749128952

VL - 47

SP - 3708

EP - 3716

JO - Investigative Ophthalmology and Visual Science

JF - Investigative Ophthalmology and Visual Science

SN - 0146-0404

IS - 9

ER -