灵芝多糖诱导耐药卵巢癌细胞凋亡的实验研究

摘要

目的 研究灵芝多糖在体外对耐药卵巢癌细胞株SKOV-3/DDP的增值抑制和诱导凋亡的作用,并初步探讨其作用机制.方法 (1)采用MTT法检测灵芝多糖对耐药卵巢癌细胞株SKOV-3/DDP细胞增殖的抑制作用.(2)采用光学显微镜、相差显微镜观察灵芝多糖作用后细胞形态变化.(3)采用DNA片段检测证实细胞凋亡;(4)采用DCFH-DA荧光分光光度法检测细胞内ROS水平.(5)采用Real-time PCR技术检测谷胱甘肽过氧化物酶(GSH-Px)和过氧化氢酶(CAT) mRNA的表达,观察细胞内过氧化物的清除情况.结果 灵芝多糖可明显抑制SKOV-3/DDP3细胞增殖并诱导其发生凋亡.结论 灵芝多糖诱导细胞凋亡的机制可能是通过增加肿瘤细胞内 ROS的生成,降低GSH-Px和 CAT mRNA 的表达,即通过氧化应激途径诱导SKOV-3/DDP细胞发生凋亡.%Objective To study in vitro of Ganoderma lucidum polysaccharides (GLPs) resistant human ovarian cancer cell line SKOV-3/DDP value-added role of inhibition and induction of apoptosis ,and to explore its mechanism .Methods (1 )using MTT assay GLPs on SKOV-3/DDP resistant ovarian cancer cell line cell proliferation . (2) using acridine orange/ethidium bromide staining of apoptotic cells detected by fluorescence microscopy .(3)detected by DNA fragments confirmed apoptosis ;(4)using DCFH-DA fluores-cence spectrophotometry to determine the level of intracellular ROS . (5) using Real-time PCR detection of glutathione peroxidase (GSH-Px) and catalase(CAT) mRNA expression was observed in the clearance of intracellular peroxides .Results SKOV-3/DDP3 GLPs can inhibit cell proliferation and induced apoptosis .Conclusion GLPs can induce apoptosis of tumor cells by increasing ROS generation ,reduced GSH-Px and CAT mRNA expression Induced by oxidative stress that SKOV-3/DDP apoptosis .