臭椿酮抑制急性骨髓性白血病细胞恶性生物学行为的研究  

Study on the Inhibition of Ailanthone on Malignant Biological Behavior of Acute Myeloid Leukemia Cells

作　　者：徐奔 覃锐 向航 许京淑 廖子龙 向金平 XU Ben;QIN Rui;XIANG Hang;XU Jingshu;LIAO Zilong;XIANG Jinping(Department of Hematology,Central Hospital of Enshi Tujia and Miao Autonomous Prefecture,Hubei Enshi 445000,China)

机构地区：[1]恩施土家族苗族自治州中心医院血液科,湖北恩施445000

出　　处：《生物技术进展》2022年第5期769-777,共9页Current Biotechnology

摘　　要：为了探讨臭椿酮(ailanthone,AIL)对急性骨髓性白血病(acute myelogenous leukemia,AML)细胞恶性生物学行为的影响,用不同浓度(0.2、0.4、0.8、1.6、3.2μmol·L^(-1))的AIL处理对数生长期的HL-60细胞,将miR-449a mimic质粒、mimic对照质粒、miR-449a inhibitor质粒、inhibitor对照质粒分别转染至未经任何处理的HL-60细胞,并用1.0μmol·L^(-1)浓度的AIL处理细胞24 h。采用CCK-8法检测细胞增殖水平,细胞划痕实验检测细胞迁移水平,Transwell小室法检测细胞侵袭水平,Annexin V-FITC/PI双染法检测细胞凋亡水平,qRT-PCR法检测miR-449a mRNA表达水平,Western blot法检测磷脂酰肌醇3-激酶(PI3K)、磷酸化PI3K(p-PI3K)、蛋白激酶B(AKT)、磷酸化AKT(p-AKT)蛋白表达水平。结果显示,AIL干预后HL-60细胞增殖抑制率、凋亡率升高,细胞迁移率及细胞侵袭数降低(P<0.05),miR-449a mRNA表达量升高(P<0.05)。过表达miR-449a可以抑制HL-60细胞增殖、迁移和侵袭,并诱导细胞凋亡(P<0.05),抑制miR-449a的表达可以起到逆转AIL抑制HL-60细胞增殖、迁移和侵袭,诱导细胞凋亡的作用(P<0.05)。AIL能够显著降低HL-60细胞中p-PI3K/PI3K和p-AKT/AKT比值(P<0.05),抑制miR-449a表达可以逆转AIL对HL-60细胞p-PI3K/PI3K和p-AKT/AKT比值的下调作用(P<0.05)。结果表明,AIL可通过上调miR-449a抑制AML细胞的增殖、迁移和侵袭,并诱导细胞凋亡,其作用机制可能与抑制PI3K/AKT信号通路有关。结果表明,AIL有望成为AML治疗的候选药物。To investigate the effect of ailanthone(AIL)on the malignant biological behavior of acute myeloid leukemia(AML)cells,HL-60 cells in logarithmic growth phase were treated with AIL of different concentrations(0.2,0.4,0.8,1.6,3.2μmol·L^(-1)),miR-449a mimic plasmid,mimic control plasmid,miR-449a inhibitor plasmid and inhibitor control plasmid were transfected into HL-60 cells without any treatment,and the cells were treated with 1.0μmol·L^(-1) AIL for 24 hours.Cell proliferation was detected by CCK-8 assay,cell migration by scratch test,cell invasion by Transwell chamber assay,cell apoptosis by Annexin V-FITC/PI double staining and miR-449a mRNA expression by qRT-PCR.Western blot was used to detect the expression of Phosphatidylino⁃sitol 3-kinase(PI3K),phosphorylated PI3K(P-PI3K),protein kinase B(AKT)and phosphorylated Akt(p-AKT).The re⁃sults showed that after the intervention of AIL,the proliferation inhibition rate and apoptosis rate of HL-60 cells were significantly increased,the cell migration rate and the number of cell invasion were significantly decreased(P<0.05),and the expression of mir-449a mRNA was significantly increased(P<0.05).Overexpression of miR-449a could inhibit the proliferation,migration and invasion of HL-60 cells and induce apoptosis(P<0.05).Inhibition of miR-449a expression could reverse the inhibitory effect of AIL on proliferation,migration and invasion of HL-60 cells and induce apoptosis(P<0.05).The ratio of p-PI3K/PI3K and p-AKT/AKT in HL-60 cells were significantly decreased by AIL(P<0.05),inhibition of miR-449a expression could reverse the down-regulation of AIL on p-PI3K/PI3K and p-AKT/AKT ratio in HL-60 cells(P<0.05).The results indicated that AIL could inhibit the proliferation,migration,invasion and induce apoptosis of AML cells by up-regulating miR-449a,its mechanism may be relat⁃ed to the inhibition of PI3K/Akt signaling pathway.AIL is expected to be a candidate drug for the treatment of AML.

关 键 词：臭椿酮 急性骨髓性白血病 miR-449a PI3K/AKT信号通路 增殖 迁移 侵袭 凋亡 

分 类 号：R733.7[医药卫生—肿瘤] Q291[医药卫生—临床医学]