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 fluorescence 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.
出处
《中国实验诊断学》
北大核心
2011年第3期413-417,共5页
Chinese Journal of Laboratory Diagnosis
