鱼藤素对淋巴瘤Daudi细胞株细胞增殖细胞凋亡的影响及其机制  

Effect of Deguelin on proliferation and apoptosis of lymphoma Daudi cells and its mechanism

作　　者：刘红利[1,2] 陈燕[1] 吴秋玲[1] 陈卫华[1] 何静[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院血液病研究所,武汉430022 [2]华中科技大学同济医学院附属协和医院肿瘤中心,武汉430022

出　　处：《中华肿瘤杂志》2007年第3期176-180,共5页Chinese Journal of Oncology

摘　　要：目的 观察鱼藤素对人Burkitt淋巴瘤Daudi细胞株细胞增殖、细胞凋亡和细胞周期的影响，并探讨其分子机制。方法 四甲基偶氮唑蓝（MTT）法检测细胞增殖活性，Hoechst 33258染色和Annexin-V／PI双标法检测细胞凋亡，流式细胞仪检测细胞周期分布，Western blot检测细胞内cyclin D1和pRb的蛋白表达。结果鱼藤素对Daudi细胞具有明显的增殖抑制作用，而对正常人外周血单个核细胞（PBMC）抑制作用不明显。鱼藤素可以诱导Daudi细胞凋亡，Hoechst 33258染色可见典型凋亡小体。AnnexinV／PI双标法显示，鱼藤素诱导细胞发生早期凋亡，并呈剂量依赖性，20、40、80nmol／L鱼藤素作用24h时，凋亡率分别为15．46％±0．62％、18．48％±2．98％和31．42％±1．43％。鱼藤素作用Daudi细胞后，主要使细胞周期聚积于G0／G1期，G0／G1期细胞比例随鱼藤素剂量增大而逐渐增高，40nmol／L鱼藤素作用24h达56．56％；相反，S期细胞比例随鱼藤素剂量增大而逐渐降低，对G2／M期细胞作用不明显。鱼藤素使cyclinD1及pRb蛋白表达降低，呈剂量依赖关系。结论 鱼藤素抑制Daudi细胞增殖，使细胞阻滞于G0／G1期，并诱导细胞凋亡。其抗肿瘤机制可能与下调cychn D1和pRb蛋白表达有关。Objective To investigate the anticancer effects and molecular mechanism of deguelin on human Burkitt＇s lymphoma Daudi cells in vitro and compare the cytotoxicity of deguelin on Daudi cells and human peripheral blood monocular cells （HPBMC）. Methods The effects of deguelin on the growth of Daudi cells were studied by 3-（4, 5-dimethyl-2-thiazolyl ）-2, 5diphenyl-2H-tetrazolium （MTY） assay. Apoptosis was assessed through Hoechst 33258 staining and annexin V/PI double-labeled cytometry. The effect of deguelin on the cell cycle of Daudi cells was studied by propidium iodide method. The expressions of cychn D1 and pRb were checked by Western blot. Results The proliferation of Daudi cells was decreased in deguelin-treated group with a 24 h ICso value of 51.55 nmol/L. Deguelin induced Daudi cells apoptosis in a time- and dose-dependent manner. Daudi ceils treated with deguehn showed an increase of Go/G1 phase and decrease of S phase. The Daudi cells treated with 0, 5, 10, 20, 40 nmol/L deguelin for 24 h, showed an increased Go/G1 phase from 37. 34% to 56.56%, whereas decreased S phase from 37.72% to 21.36%, respectively. PBMC was less sensitive to the cytotoxic effect of deguelin than Daudi cells. The expression of cyclin D1 and pRb protein were decreased in Daudi cells treated with deguelin. Conclusion Deguelin can inhibit the proliferation of Daudi cells by regulating the cell cycle that arrest cells at Go/G1 phase and inducing apoptosis. Moreover, deguelin demonstrats low toxicity in PBMC but selectively induces apoptosis of Daudi cells. The antitumor effects of deguehn may be related to down-regulation of the expression of cyclin D1 and pRb protein.

关 键 词：鱼藤素 BURKITT淋巴瘤 CYCLIN D1 PRB 细胞凋亡 

分 类 号：R686[医药卫生—骨科学]