Genistein抑制人卵巢癌细胞系SKOV_3增殖和诱导凋亡发生的研究  

Proliferation Inhibition and Apoptosis Onset in Human Ovarian Carcinoma Cell Line SKOV_3 Induced by Genistein

作　　者：李昱[1] 米粲[1] 

机构地区：[1]重庆医科大学病理学教研室,重庆400016

出　　处：《癌症》2003年第6期586-591,共6页Chinese Journal of Cancer

摘　　要：背景与目的:许多研究表明Genistein对多种肿瘤细胞有抑制作用,但有关Genistein对卵巢癌细胞作用的报道很少。本研究旨在通过观测Genistein对人卵巢癌细胞系SKOV3的抑制增殖和诱导凋亡作用,探讨其抗癌作用的机理。方法:应用四甲基偶氮唑盐(MTT)法检测不同浓度Genistein对SKOV3的生长抑制作用;吖啶橙/溴乙锭(AO/EB)荧光染色法及电镜观察凋亡细胞及凋亡小体;流式细胞仪分析细胞周期及凋亡率;琼脂糖凝胶电泳检测凋亡特征性DNA梯形带;免疫细胞化学李昱,等.Genistein抑制人卵巢癌细法及RT-PCR法分别检测细胞增殖凋亡调控相关蛋白及其mRNA的表达。结果:Genistein对SKOV3细胞的增殖抑制作用呈时间及浓度依赖性,20μmol/L和40μmol/LGenistein作用72h后,细胞的生长抑制率达72.07%和74.93%。20μmol/LGenistein作用SKOV3细胞48h后出现细胞周期的G2/M期阻滞。荧光显微镜和电镜均观察到用药后凋亡细胞典型的形态学特征。细胞凋亡率以Genistein20μmol/L组最高,达23.7%。凝胶电泳观察到特征性DNA梯形带。20μmol/LGenistein作用SKOV348h后,bcl-2基因表达水平降低,而p21WAF1/CIP1和bax基因表达增加(P<0.05);PCNA、Bcl-2及cyclinB1蛋白表达水平降低,Bax、p21WAF1/CIP1蛋白表达增加(P<0.01)。BACKGROUND &OBJECTIVE:It was reported that Genistein possess inhibitory effects on vario us carcinomas.But the report on the e ffects of Genistein on human ovarian carcinom a was rare.This study was aimed to investig ate the proliferation inhi bition and apoptosis onset in human o varian carcinoma cell lines SKOV 3 induced by Genistein and to discuss i ts antitumor mechanism.METHODS :Inhibitory effect of Genistein on SK OV 3 proliferation was tested by MTT method.Apoptotic cell and apoptotic bodies were observed b y acridine orang e /ethidium bromide (AO /EB)staining and electron microscope (EM).Cell apoptosis percentag e and cell cycle phase distribution were measured by flow cytometric assay(FCM).The characteristic DNA ladders of apoptosis were determined by ag aros e g el electrophoresis(AGE).The expression of proliferation and apoptosis asso ciated proteins and their mRNA were determined using immunocytochemis try and RT-PCR.RESULTS :A time-dependent and dose-dependent proliferation inhibition of Genistein wa s demonstrated in SKOV 3 ;the prominent inhibitory concentr ations were 20μmol /L and 40μmol /L(P<0.01)and the 72 nd hour was the most apparent phase(P<0.01).The g rowth inhibition rates of the c ancer cells reached 72.07%and74.93%.A G 2 /M cell cycle arrest was induced in SK OV 3 exposed to 20μmol /L Genistein.The characteristic morp holog ical chang es of apoptosis were observed by AO /EB staining and EM in SKOV 3 exposed to Genistein.The apoptosis rate was hig hest at 20μmol /L concentration,reached 23.7%.DNA ladder appeared by AGE.The expression of bcl-2mRNA w as decreased,p21 WAF1/CIP1 and bax mRNA was increased(P<0.05),the expression of PCNA,Bcl-2,and c yclin B 1 protein were decreased,Bax and p21 WAF1/CIP1 protein were increased in SKOV 3 exposed to 20μmol /L Genistein for 48hours(P<0.01).CONCLUSION:Genistein could inhibit proliferat ion and induce apoptosis in ovarian c arcinoma cell lines SKOV 3 .Its inhibitory e

关 键 词：GENISTEIN 卵巢癌 细胞增殖 细胞凋亡 血管生成 PCNA 

分 类 号：R737.31[医药卫生—肿瘤]