醋酸棉酚诱导多发性骨髓瘤细胞凋亡及其机制的实验研究

Apoptosis of Multiple Myeloma Cells Induced by Gossypol Acetic Acid In Vitro and Its Mechanism

本研究探讨醋酸棉酚对经典人多发性骨髓瘤细胞系RPMI8226的作用及其机制。采用四唑盐比色试验(MTT)、细胞形态学观察、流式细胞仪检测、DNA电泳、Western-blot等方法分别观察醋酸棉酚对RPMI8226细胞的抑制增殖、诱导凋亡效应及其机制。结果表明:醋酸棉酚在>16μmol/L浓度时可显著抑制RPMI8226细胞的增殖、促进其凋亡,随着剂量的增加以及时间的延长,效应越明显。研究还发现,醋酸棉酚是通过抑制线粒体膜电位(Δψm)、诱导Bax、Caspase-3等凋亡相关蛋白的表达来实现上述效应的。结论:醋酸棉酚对多发性骨髓瘤RPMI8226细胞具有选择性抑制增殖、促进凋亡的效应。

This study was aimed to investigate the apoptosis effect of gossypol acetic acid on classic human multiple myeloma RPMI8226 cell line in vitro and its mechanism. The inhibitory effect on proliferation of RPMI8226 cells was evaluated by means of MTT assay. Cytotoxic effect and apoptosis was identified and analyzed with the aid of transmission electron microscopy, mitochondria membrane potential （MMP） and DNA gel electrophoresis. Meanwhile, Western-blot assay was used to detect the changes of several key cell apoptosis regulatory proteins such as BAd（, caspase- 3 and caspase-8 in these cells before and after treatment. The results showed that low concentrations of gossypol acetic acid （ 〉 16 μmol/L） could suppress the proliferation and induce the apoptosis in RPMI8226 cells effectively. At the same time, gossypol acetic acid could also down-regulate the mitochondrial membrane potential, up-regulate the expression of the apoptosis-related protein such as BAX and caspase-3. It is concluded that the gossypol acetic acid can selectively induce proliferation inhibition and apoptosis of multiple meleloma RPMI8226 cells with a smaller dose.