芒柄花黄素对白血病HL-60细胞增殖、凋亡和miR-21/PTEN/AKT信号通路的影响  

Effects of formononetin on proliferation,apoptosis and miR-21/PTEN/AKT signaling pathway of HL-60 cells

作　　者：袁长威 熊媛媛 张青兰 刘新建 魏旭东 宋永平 Yuan Changwei;Xiong Yuanyuan;Zhang Qinglan;Liu Xinjian;Wei Xudong;Song Yongping(Department of Oncology and Hematology,Cancer Hospital Affiliated to Zhengzhou University,Henan Zhengzhou 450003,China)

机构地区：[1]郑州大学附属肿瘤医院肿瘤血液科,河南郑州450003

出　　处：《现代肿瘤医学》2020年第7期1062-1066,共5页Journal of Modern Oncology

基　　金：国家自然科学基金资助项目(编号:81470287)。

摘　　要：目的:探讨芒柄花黄素对白血病HL-60细胞增殖、凋亡的影响及其机制。方法:以不同浓度(0、12.5、25、50、100和200μg/ml)芒柄花黄素处理HL-60细胞24、48和72 h后,采用MTT法检测细胞存活率,并计算出IC50以筛选出合适的作用浓度。采用流式细胞仪检测芒柄花黄素对HL-60细胞周期和凋亡的影响,实时荧光定量PCR检测芒柄花黄素对HL-60细胞中miR-21表达的影响,免疫印迹法检测芒柄花黄素对HL-60细胞中PTEN、AKT和p-AKT蛋白表达的影响。结果:与0μg/ml处理组相比,25、50、100和200μg/ml芒柄花黄素处理2 4 h后HL-6 0细胞的存活率明显降低(P <0. 05);同时,12. 5、25、50、100和200μg/ml芒柄花黄素分别处理48 h和72 h后HL-60细胞的存活率明显低于0μg/ml处理组(P <0. 05);芒柄花黄素作用HL-60细胞24、48和72 h后的IC50值分别为146. 50μg/ml、120. 10μg/ml和89. 00μg/ml。50、100和150μg/ml芒柄花黄素处理后HL-60细胞在G0/G1期的百分比、细胞凋亡率和PTEN蛋白的表达水平均逐渐升高,而细胞在S期和G2/M期的百分比、miR-21的表达水平以及p-AKT蛋白的表达水平均逐渐降低,且与0μg/ml相比,差异均有统计学意义(P <0. 05),但HL-60细胞中AKT蛋白的表达水平与0μg/ml组比较差异无统计学意义(P> 0. 05)。结论:芒柄花黄素可抑制白血病HL-60细胞增殖并诱导细胞凋亡,其作用机制可能与调控miR-21/PTEN/AKT信号通路有关。Objective:To investigate the effect of formononetin on the proliferation and apoptosis of HL-60 cells and its mechanism.Methods:HL-60 cells were treated with different concentrations(0,12.5,25,50,100 and 200 μg/ml) of cantharidin for 24,48 and 72 hours.The cell survival rate was measured by MTT method,and IC50 was calculated to select the appropriate concentration.The effects of formononetin on the cell cycle and apoptosis of HL-60 cells were detected by flow cytometry.The effect of formononetin on the expression of miR-21 in HL-60 cells was measured by real-time fluorescence quantitative PCR,and the effect of formononetin on the expression of PTEN,AKT and p-AKT in HL-60 cells was tested by Western blotting.Results:Compared with 0 μg/ml treatment group,the survival rate of HL-60 cells treated with 25,50,100 and 200 μg/ml formononetin for 24 hours was significantly lower(P<0.05).At the same time,the survival rate of HL-60 cells treated with 12.5,25,50,100 and 200 μg/ml formononetin for 48 hours and 72 hours was significantly lower than that of 0 μg/ml treatment group(P<0.05).The IC50 values of HL-60 cells treated with formononetin after 24,48 and 72 hours were 146.50 μg/ml,120.10 μg/ml and 89.00 μg/ml respectively.After treatment with 50,100 and 150 μg/ml of formononetin,the percentage of HL-60 cells in G0/G1 phase,apoptosis rate and PTEN protein expression level increased gradually,while the percentage of HL-60 cells in S phase and G2/M phase,miR-21 expression level and p-AKT protein expression level decreased gradually,and the difference was statistically significant compared with 0 μg/ml(P<0.05),but there was no significant difference about AKT protein expression compared with 0 μg/ml group(P>0.05).Conclusion:Formononetin can inhibit the proliferation of HL-60 cells and induce apoptosis,and its mechanism may be related to the regulation of miR-21/PTEN/AKT signaling pathway.

关 键 词：白血病 芒柄花黄素 细胞增殖 细胞周期 细胞凋亡 miR-21/PTEN/AKT信号通路 

分 类 号：R733.7[医药卫生—肿瘤]