汉黄芩素抑制PI3K/Akt/mTOR信号通路影响急性白血病HL-60细胞增殖  

Effect of Wogonin on the Proliferation in Acute Myeloid Leukemia HL-60 Cell Through Inhibition of PI3K/Akt/mTOR Signaling Pathway

作　　者：高平章 韩旭花 白赛赛 谢晓兰 

机构地区：[1]泉州师范学院化工与材料学院,福建泉州362000

出　　处：《泉州师范学院学报》2017年第6期1-5,共5页Journal of Quanzhou Normal University

基　　金：福建省自然科学基金项目(2017J01856);国家级大学生创新创业训练项目(201410399006);泉州市科技计划项目(2011Z24)

摘　　要：探讨汉黄芩素对急性白血病HL-60细胞增殖及其可能的作用机制.以不同浓度(10,20,40,80,100μmol/L)的汉黄芩素分别作用于HL-60 24,48,72h后,采用MTT法检测其对细胞增殖活性的影响;台盼蓝拒染法观察汉黄芩素作用HL-60细胞48h后,对细胞活力的影响;Western blot法检测汉黄芩素对HL-60细胞中PI3K/AKT/mTOR和Raf/MEK/ERK信号通路中相关蛋白的表达.结果表明:汉黄芩素能够显著抑制HL-60细胞增殖,具有浓度依赖和时间依赖性.此外,汉黄芩素对PI3K/AKT/mTOR信号通路中p-PI3K、pAKT、mTOR和p-mTOR蛋白表达具有明显的抑制作用;对Raf/MEK/ERK信号通路中的MEK、ERK蛋白表达增加,抑制ERK1/2的磷酸化水平.汉黄芩素抑制HL-60细胞增殖和逆转耐药性,其作用机制可能与抑制PI3K/AKT/mTOR信号通路蛋白的表达和下调Raf/MEK/ERK信号通路中ERK磷酸化水平有关.The aim of this study is to investigate the inhibitory effect of wogonin on the proliferation of acute myeloid leukemia HL-60 cell and its underlying mechanism.HL-60 cells were treated with wogonin at different concentrations of 10-100μmol/L for 24-72 hours,respectively.MTT assay was performed to assess the cell proliferation.Trypan blue exclusion assay was used to determine cell viability.Western blot was adopted to analyze the cell proliferation related proteins expression of PI3 K/AKT/mTOR and Raf/MEK/ERK signaling pathway in HL-60 cells.The results showed that Wogonin obviously inhibited the viability of HL-60 cells in a dose dependent and time dependent manner,the IC50 value of wogonin for 24,48,72 hours was 40,30,15μmol/L,respectively.In addition,wogonin reduced the phosphorylation of PI3 K,AKT,mTOR in the HL-60 cells.Furthermore,wogonin suppressed of mTOR in the HL-60 cells to a varying degree which showed obvious concentration-dependent manner.In Raf/MEK/ERKsignaling pathway,wogonin increased the expression of MEK and ERK,but reduced the phosphorylation of ERK.In summary,our study results indicated that wogonin inhibited proliferation and reversed drug resistance,which were mediated by the inhibition of PI3 K/AKT/mTOR signaling pathway and down-regulation the phosphorylation of ERK inRaf/MEK/ERKsignaling pathway.

关 键 词：汉黄芩素 HL-60细胞 细胞增殖 PI3K/Akt/mTOR信号通路 耐药性 Raf/MEK/ERK信号通路 

分 类 号：R966[医药卫生—药理学]