阿魏酸对U937细胞生长的抑制作用及其相关机制研究  

Inhibition of Ferulic Acid on U937 Cell Growth and Its Related Mechanism

作　　者：郑佳 吴翠翠 刘玮 姜习新 罗岚[5] 邓益媛[5] 李光[5] ZHENG Jia;WU Cui-Cui;LIU Wei;JIANG Xi-Xin;LUO Lan;DENG Yi-Yuan;LI Guang(Department of Basic Medicine,Yueyang Vocational and Technical College;Department of Hematology,Yueyang 414000,Hunan Province,China;Department of Pathology,Yueyang 414000,Hunan Province,China;Department of Laboratory Medicine,Yueyang People′s Hospital,Yueyang 414000,Hunan Province,China;Medical School of Yueyang Vocational and Technical College,Yueyang 414000,Hunan Province,China)

机构地区：[1]岳阳职业技术学院基础医学部 [2]岳阳市人民医院血液内科,湖南岳阳414000 [3]岳阳市人民医院病理科,湖南岳阳414000 [4]岳阳市人民医院检验科,湖南岳阳414000 [5]岳阳职业技术学院医学院,湖南岳阳414000

出　　处：《中国实验血液学杂志》2023年第5期1278-1283,共6页Journal of Experimental Hematology

基　　金：湖南省教育厅科学研究项目立项课题(21C1540)。

摘　　要：目的:探讨阿魏酸(FA)对人急性髓系白血病(AML)细胞系U937细胞增殖、凋亡及Toll样受体4(TLR4)/核因子κB(NF-κB)信号通路的影响。方法:使用不同浓度的FA(0、10、25、50、100、200μmol/L)分别处理人AML细胞系Kasumi-1、HL-60、U937细胞24 h后,采用CCK-8法检测细胞存活率,计算各个细胞系的半数抑制浓度(IC_(50))。将U937细胞分为对照组、FA组(50μmol/L FA)、FA+pcDNA组(50μmol/L FA+转染空载质粒)、FA+pcDNA-TLR4组(50μmol/L FA+转染TLR4过表达质粒)。采用流式细胞术检测各组U937细胞周期与细胞凋亡;平板克隆形成实验检测U937细胞克隆形成能力;荧光定量PCR检测U937细胞TLR4、NF-κB p65 mRNA水平;蛋白印迹法检测U937细胞细胞周期蛋白D1(CyclinD1)、细胞周期蛋白E(CyclinE)、Bcl-2相关X蛋白(Bax)、B淋巴细胞瘤-2(Bcl-2)、半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)、TLR4、NF-κB p65蛋白表达。结果:随着FA浓度的升高,Kasumi-1、HL-60、U937细胞存活率逐渐下降(r=-0.919,r=-0.909,r=-0.900),U937细胞的IC_(50)为50.25±2.23μmol/L。与对照组比较,经药物处理U937细胞后,FA组G_(0)/G_(1)期细胞比例、细胞凋亡率、Bax、Caspase-3蛋白表达水平均显著升高(P<0.05),细胞克隆形成数、S期和G_(2)/M期细胞比例、CyclinD1、CyclinE、Bcl-2蛋白及TLR4、NF-κB p65 mRNA和蛋白表达水平均显著降低(P<0.05);与FA组和FA+pcDNA组比较,FA+pcDNA-TLR4组G_(0)/G_(1)期细胞比例、细胞凋亡率、Bax、Caspase-3蛋白表达水平均显著降低(P<0.05),细胞克隆形成数、S期和G_(2)/M期细胞比例、CyclinD1、CyclinE、Bcl-2蛋白及TLR4、NF-κB p65 mRNA和蛋白表达水平均显著升高(P<0.05)。结论:FA通过抑制TLR4/NF-κB信号通路抑制U937细胞增殖,促进细胞凋亡。Objective:To investigate the effects of ferulic acid(FA)on proliferation,apoptosis and Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)signaling pathway of human acute myeloid leukemia(AML)U937 cells.Methods:Different concentrations of FA(0、10、25、50、100、200μmol/L)was used to treat human AML cell lines Kasumi-1,HL-60,and U937 cells respectively for 24 h.The cell survival rate was detected by cell counting kit-8 method,and the 50%inhibitory concentration(IC_(50))of each cell line was calculated.The U937 cells were divided into control group,FA group(50μmol/L FA),FA+pcDNA group(50μmol/L FA+transfected with empty plasmid),FA+pcDNA-TLR4 group(50μmol/L FA+transfected with TLR4 overexpression plasmid).Flow cytometry was used to detect U937 cell cycle and cell apoptosis;plate clone formation test was used to detect U937 cell clone formation ability;fluorescence quantitative PCR was used to detect the TLR4,NF-κB p65 mRNA levels in U937 cells;Western blot was used to detect the expression levels of CyclinD1,CyclinE,Bcl-2 related X protein(Bax),B cell lymphoma-2(Bcl-2),Caspase-3,TLR4,and NF-κB p65 protein in U937 cells.Results:With the increase of FA concentration,the survival rates of Kasumi-1,HL-60 and U937 cells gradually decreased(r=-0.919,r=-0.909,r=-0.900),the IC_(50)of U937 cells was 50.25±2.23μmol/L.Compared with the control group,after drug treatment of U937 cells,the ratio of G 0/G1 phase cells,apoptosis rate,expression levels of Bax and Caspase-3 proteins in FA group were significantly increased(P<0.05),the number of cell clones,the ratios of S phase and G2/M phase cells,expression levels of CyclinD1,CyclinE and Bcl-2 proteins,and TLR4,NF-κB p65 mRNA and protein were significantly decreased(P<0.05);compared with FA group and FA+pcDNA group,the ratio of G0/G1 phase cells,apoptosis rate,expression levels of Bax and Caspase-3 proteins in FA+pcDNA-TLR4 group were significantly decreased(P<0.05),the number of cell clones,the ratios of S phase and G2/M phase cells,expression levels of expression

关 键 词：阿魏酸 急性髓系白血病 U937细胞 增殖 凋亡 Toll样受体4/核转录因子-κB信号通路 

分 类 号：R733.71[医药卫生—肿瘤]