摘要
目的:研究桃儿七根醇提物对人乳腺癌MCF-7细胞增殖和凋亡的影响,探讨其抗癌作用的机制。方法:应用MTT法检测桃儿七根醇提物对人乳腺癌MCF-7细胞的生长抑制作用,倒置显徽镜、HE染色及电镜观察细胞形态学改变,流式细胞仪(FCM)分析细胞周期及凋亡率,免疫细胞化学法(ICC)检测细胞增殖、凋亡相关调控蛋白的表达。结果:桃儿七根醇提物可明显抑制MCF-7细胞的增殖,1,2,3 mg.L^(-1)桃儿七根醇提物作用72h后,细胞生长抑制率分别为43.0%,54.9%,78.5%,抑制作用呈时间及浓度依赖性。形态学观察到用药后凋亡细胞典型的形态学特征,伴有多核细胞形成。FCM分析发现桃儿七根醇提物阻断MCF-7细胞生长于细胞周期的G_2/M期,凋亡诱导作用呈时间及浓度依赖性。免疫细胞化学结果显示,2mg·L^(-1)桃儿七根醇提物作用48h后,PCNA,Bcl-2蛋白表达较对照组显著下降;而p21^(WAFI-CIP1)和c-Myc蛋白显著增加(P<0.01)。结论:桃儿七根醇提物可能通过增加p21^(WAFI-CIP1)蛋白表达及降低PCNA蛋白表达而抑制人乳腺癌MCF-7细胞的增殖,并可能下调Bcl-2蛋白表达和上调c-Myc蛋白表达来促进MCF-7细胞凋亡。
Objective: To investigate the effects and mechanism of ethanol extracts from the roots of Podophyllum emodi var. chinense on proliferation and apoptosis of breast carcinoma cell line MCF-7. Methods: Inhibition of the MCF-7 proliferation by the ethanol extracts from P. emodi var. chinense roots was evaluated using a MTT assay. The morphodifferentiation of the MCF-7 was studied by reversal micro- scope, electron microscope and HE staining. The cellular apoptosis rate and cell cycle phase distribution of the MCF-7 were measured by a flow cytometric assay (FCM). The expressions of proteins modulating the proliferation and apoptosis were analyzed by an immunocytochemical assay (ICC). Results: The ethanol extracts suppressed the MCF-7 proliferation in a time-dependent and dose-dependent manner, showing the inhibition rate of 43.0% (1mg·L^-1), 54.9% (2mg·L^-1) and78.5% (3mg·L^-1) after the culture of the MCF-72 together with the extracts for 72 hours. The morphological study found that the multinucleate giant cells existed in the apoptotic cells after the culture of the MCF-72 with 2 mg·L^-1 of the ethanol extracts. The FCM assay showed that the ethanol extracts induced a G2/M cell cycle arrest in a time- and dose-dependent manner. The ICC assay disclosed that the protein expressions of PCNA, Bcl- 2 significantly decreased (P 〈0.01 ), and p21^WAF1/CIPI and c-Myc significantly increased (P 〈0.01 ) after the culture of the MCF-7 with the ethanol extracts 2 mg.L^-1 for 48 hours. Conclusion: The ethanol extracts inhibited the MCF-7 proliferation by up-regulation of p21^WAF1/GIP1 level and down-regulation of PCNA level. The apoptosis was produced by down-regulation of Bcl-2 expression and up-regulation of c-Myc proteins expression.
出处
《中国新药杂志》
CAS
CSCD
北大核心
2006年第13期1064-1067,共4页
Chinese Journal of New Drugs
基金
青海省科技厅院士基金(2001-01-J-801)
关键词
桃儿七根醇提物
人乳腺癌细胞系MCF-7
细胞增殖
细胞凋亡
ethanol extracts from the root of P, emodi var. chinense
human breast carcinomacell lines MCF-7
proliferation
apoptosis