Department of Neurosurgery, Otto von Guericke University, Magdeburg, Germany.
Onco Targets Ther. 2013 Sep 16;9:1269-76. doi: 10.2147/OTT.S45154. eCollection 2013.
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.
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).
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.
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.
白藜芦醇是一种植物雌激素，具有多种抗增殖和促凋亡作用。本体外研究旨在分析白藜芦醇对大鼠 GH3 垂体腺瘤细胞活力和凋亡调节剂表达的影响。
将 GH3 细胞用浓度为 20 至 100μM 的白藜芦醇孵育 48-72 小时。用血细胞计数器定量测定细胞活力。我们通过核小体释放的酶联免疫吸附测定（ELISA）和 DNA 降解（一维凝胶电泳）评估白藜芦醇杀死 GH3 细胞的能力。使用定量实时聚合酶链反应（qRT-PCR）测量存活素、B 细胞淋巴瘤-2 蛋白（BCL-2）和 BCL-2 相关 X 蛋白（BAX）的相对信使 RNA（mRNA）表达，并归一化为β2 微球蛋白。
随着白藜芦醇浓度的增加，GH3 细胞的存活率显著降低。在用 100μM 白藜芦醇处理的 GH3 细胞中，ELISA 显示核小体释放显著增加，这通常发生在细胞凋亡过程中。同时，凝胶电泳显示 DNA 降解为随机片段，表明大多数 GH3 细胞发生坏死性细胞死亡模式。在用 100μM 白藜芦醇处理的 GH3 细胞中，qRT-PCR 检测到 BCL-2 mRNA 表达显著降低，存活素 mRNA 表达降低，而 BAX mRNA 表达变化未发现。白藜芦醇处理后，GH3 细胞中的 BAX/BCL-2 比值显著增加。
白藜芦醇通过诱导非凋亡性细胞死亡和凋亡以剂量依赖性方式降低 GH3 细胞活力。GH3 细胞中的凋亡可能是由白藜芦醇依赖性凋亡抑制剂（即 BCL-2 和可能是存活素）下调介导的。进一步研究白藜芦醇对垂体腺瘤细胞的潜在影响是有必要的。
