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 M.
AU - Polans, Arthur S.
PY - 2006/9
Y1 - 2006/9
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
SN - 0146-0404
VL - 47
SP - 3708
EP - 3716
JO - Investigative Ophthalmology and Visual Science
JF - Investigative Ophthalmology and Visual Science
IS - 9
ER -