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Resveratrol decreases B-cell lymphoma-2 expression and viability in GH3 pituitary adenoma cells of the rat

Authors Voellger B, Kirches E, Wilisch-Neumann A, Weise A, Tapia-Perez JH, Rupa R, Mawrin C, Firsching R

Received 13 March 2013

Accepted for publication 24 July 2013

Published 16 September 2013 Volume 2013:6 Pages 1269—1276

DOI https://doi.org/10.2147/OTT.S45154

Checked for plagiarism Yes

Review by Single anonymous peer review

Peer reviewer comments 4

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Benjamin Voellger,1 Elmar Kirches,2 Annette Wilisch-Neumann,2 Andreas Weise,1 Jorge Humberto Tapia-Perez,1 Rosita Rupa,1 Christian Mawrin,2 Raimund Firsching1

1Department of Neurosurgery, 2Department of Neuropathology, Otto von Guericke University, Magdeburg, Germany

Objective: Resveratrol is a phytoestrogen with various antiproliferative and proapoptotic effects. This in vitro study aimed to analyze the effect of resveratrol on the viability and expression of modulators of apoptosis in GH3 pituitary adenoma cells of the rat.
Methods: GH3 cells were incubated with resveratrol concentrations from 20 to 100 µM for 48–72 hours. Cell viability was quantified using a hemocytometer. We assessed the ability of resveratrol to kill GH3 cells by an enzyme-linked immunosorbent assay (ELISA) of nucleosome liberation and by DNA degradation (unidimensional gel electrophoresis). Relative messenger RNA (mRNA) expression of survivin, B-cell lymphoma-2 protein (BCL-2) and BCL-2-associated X protein (BAX) normalized to β2 microglobulin was measured using quantitative real-time polymerase chain reaction (qRT-PCR).
Results: GH3 cell survival significantly decreased with increasing concentrations of resveratrol. In GH3 cells treated with 100 µM resveratrol, ELISA demonstrated a significant rise of nucleosome liberation, which typically occurs during apoptosis. In parallel, gel electrophoresis showed degradation of DNA into random fragments, pointing to a necrotic mode of cell death in most GH3 cells. In GH3 cells treated with 100 µM resveratrol, qRT-PCR detected a significant decrease of BCL-2 mRNA expression and a decrease of survivin mRNA expression, whereas a change of BAX mRNA expression could not be found. The BAX/BCL-2 ratio was significantly increased in GH3 cells after resveratrol treatment.
Conclusions: Resveratrol reduces GH3 cell viability in a dose-dependent manner by inducing nonapoptotic cell death and apoptosis. Apoptosis in GH3 cells is probably mediated by resveratrol-dependent downregulation of apoptosis inhibitors, namely BCL-2 and possibly survivin. Further investigation of the potential effects of resveratrol on pituitary adenoma cells is warranted.

Keywords: resveratrol, phytoestrogens, viability, GH3 cells

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