摘要
目的:探讨芒柄花素对人非小细胞肺癌(NSCLC)细胞增殖、凋亡的影响,并探讨其作用机制。方法:培养人NSCLC细胞株A549,分别按10,20,40,80,160μmol·L^(-1)梯度浓度加入芒柄花素,另设不含药物空白组,培养48 h后采用噻唑蓝(MTT)比色法检测细胞增殖活力,利用Annexin V-FIFC/碘化丙啶(PI)双标法检测细胞凋亡情况,利用流式细胞仪检测细胞周期,利用实时荧光定量PCR(Real-time PCR)和免疫印迹法(Western blot)分别检测细胞中周期蛋白E_1(cyclin E_1),B淋巴细胞瘤-2(Bcl-2)和Bcl-2相关X蛋白(Bax)mRNA和蛋白表达。结果:40,80,160μmol·L^(-1)芒柄花素组细胞生长抑制率和细胞凋亡率均高于空白组(P<0.05)。与空白组比较,40,80,160μmol·L^(-1)芒柄花素组G_0/G_1期细胞比例上升,S期细胞比例下降,80,160μmol·L^(-1)芒柄花素组下降更为明显(P<0.05);40,80,160μmol·L^(-1)芒柄花素组cyclin E_1,Bcl-2 mRNA和蛋白表达量均较空白组降低,Bax mRNA和蛋白表达量升高,与40μmol·L^(-1)芒柄花素组比较,80,160μmol·L^(-1)芒柄花素组cyclin E_1,Bcl-2 mRNA和蛋白表达量均降低,Bax mRNA和蛋白表达量升高(P<0.05)。结论:芒柄花素可抑制NSCLC细胞增殖,加速细胞凋亡发生,可能通过下调cyclin E_1表达而影响细胞周期,并通过调控Bcl-2和Bax表达而促使细胞凋亡发生。
Objective： To investigate the effects of formononetin on proliferation and apoptosis of human non-small cell lung cancer cells and its relevant mechanism. Method： The human non-small cell lung cancer（ NSCLC） cell line A549 was cultured,and added with formononetin with the concentrations of 10,20,40,80 and160 μmol·L^-1. The blank group was only added PBS. After cultured for 48 h,the cell proliferations were detected by using methylthiazolyldiphenyl-tetrazolium bromide（ MTT） assay, the cell apoptosis were tested by using Annexin V-FITC/propidium iodide（ PI） double staining,the cell cycles were detected by using flow cytometry,the expressions of cyclin E1,Bcl-2 and Bax mRNA and proteins were detected by using Real-time PCR and Western blot method. Result： The cell growth inhibition rates and the cell apoptosis rates at the concentrations of 40,80 and 160 μmol·L^-1of formononetin were higher than the blank group（ P〈0. 05）. Compared with the blank group,the proportions of G0/G1 phase at the concentrations of 40,80 and 160 μmol·L^-1of formononetin were increased,while the proportions of S phase were decreased,especially the concentrations of 80 and 160 μmol·L^-1of formononetin（ P〈0. 05）. The gene and protein expressions of cyclin E1 and Bcl-2 at the concentrations of 40,80 and 160 μmol·L^-1were lower than the blank group,while the mRNA and protein expressions of Bax were increased; compared with 40 μmol·L^-1of formononetin,the expression levels of cyclin E1 and Bcl-2 mRNA and protein expressions were decreased at the concentrations of 80,160 μmol·L^-1of formononetin,but with increase in Bax mRNA and protein expressions（ P〈0. 05）. Conclusion： Formononetin could inhibit the proliferation of NSCLC cells,and accelerate cell apoptosis. It might affect the cell cycle by down-regulating cyclin E1 expression,and promote cell apoptosis by regulating the expressions of Bcl-2 and Bax.
作者
李自全
孟祥娇
LI Zi-quan MENG Xiang-jiao(The Second People＇ s Hospital of Sichuan, Chengdu 610041 , China Shandong Academy of Medical Sciences, Ji＇ nan 250062, China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2016年第20期138-142,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81301868)
关键词
芒柄花素
人非小细胞肺癌
细胞增殖
细胞凋亡
机制
formononetin
human non-small cell lung cancer
cell proliferation
cell apoptosis
mechanism
