摘要
目的 研究genistein对人卵巢癌细胞系 3AO抑制增殖和促进凋亡的作用 ,探讨其抗癌作用的机制。方法 应用MTT法检测genistein对 3AO的生长抑制作用 ,电镜观察凋亡细胞及凋亡小体 ,FCM分析细胞周期及凋亡率 ,免疫细胞化学法检测细胞增殖凋亡调控相关蛋白的表达。结果 genistein可明显抑制 3AO的增殖 ,且这种抑制作用呈时间及浓度依赖性。FCM分析发现 genistein阻断 3AO细胞生长于细胞周期的G2 /M期 ,且 2 4～ 72h可见明显亚G1峰。电镜观察到用药后凋亡细胞典型的形态学特征。免疫细胞化学结果显示 ,2 0 μmol/L的genistein作用 4 8h后 ,PCNA、bcl 2及CyclinB1蛋白表达降低 ,而p2 1WAF1/CIP1和bax及CyclinB1蛋白表达增加。结论 genistein对卵巢癌细胞系 3AO的生长有明显的呈时间及浓度依赖性的抑制作用 ,其阻断细胞的生长主要在细胞周期的G2 /M期 ,且这种生长抑制作用与 p2 1WAF1/CIP1蛋白水平表达增加及PCNAcylinB1蛋白表达降低有关 ,且可通过下调bcl 2蛋白表达 ,上调bax蛋白表达诱导卵巢癌细胞凋亡。
Objective To investigate the effects and mechanism of genistein on proliferation inhibition and onset of apoptosis in ovarian carcinoma cell line 3AO. Methods Antiproliferative effect of genistein against 3AO was tested by MTT method. Apoptotic phenotype of 3AO was observed by electron microscope. Cell apoptosis percentage and cell cycle phase distribution of 3AO were measured by flow cytometric assay. The expression of proliferation and apoptosis associated protein was determined by immunocytochemical method. Results A time-dependent and dose-dependent proliferation inhibition was demonstrated in 3AO, and genistein induced a G 2/M cell cycle arrest . The characteristic morphological changes of apoptosis in 3AO cells was observed after treated by genistein. 20 μmol/L genistein could down-regulate the expression of PCNA, cyclin B 1 and bcl-2; and up-regulate the expression of p21 WAF1/CIP1 and bax. Conclusion The time-dependent and dose-dependent proliferation inhibitory effect of genistein on ovarian cancer cells appears to be due to the up-regulation of p21 WAF1/CIP1 and PCNA expression, down-regulation of Cyclin B 1 expression, and the apoptosis is related to the up-regulation of apoptotic proteins bax and down-regulation of anti-apoptotic proteins bcl-2. The results provide evidence for the potential usefulness of genistein in the prevention and treatment of human ovarian carcinoma.
出处
《肿瘤》
CAS
CSCD
北大核心
2003年第6期490-493,共4页
Tumor
