过表达miR-186对白血病HL-60细胞增殖和凋亡作用机制的研究  

Study on the regulation of miR-186 overexpression on proliferation and apoptosis of leukemia cells

作　　者：冯永笑[1] 赵晓亮 张丑丑[1] 董艳琴[1] Feng Yongxiao;Zhao Xiaoliang;Zhang Chouchou;Dong Yanqin(Department of Integrated Chinese and Western Medicine,Gansu Cancer Hospital,Lanzhou 730050,China;Department of Imaging,Linxia People's Hospital,Linxia 731100,Gansu,China)

机构地区：[1]甘肃省肿瘤医院中西医结合科,甘肃兰州730050 [2]临夏市人民医院影像科,甘肃临夏731100

出　　处：《兰州大学学报（医学版）》2023年第2期26-32,共7页Journal of Lanzhou University（Medical Sciences）

基　　金：甘肃省中医药管理局科研课题资助项目(GZK-2014-78)。

摘　　要：目的探究过表达miR-186对白血病细胞增殖、凋亡及磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)信号通路的调控作用。方法选取人白血病HL-60细胞,通过实时荧光定量PCR(RT-qPCR)、细胞计数试剂-8(CCK-8)、Hoechst33258染色、酶联免疫吸附试验(ELISA)及蛋白质印迹法(WB)法分析细胞形态、miR-186相对表达量、增殖能力、凋亡情况、相关因子表达水平。结果转染24 h后,与mimic NC组相比,miR-186组细胞生长受到抑制,miR-186相对表达量、凋亡细胞数、超氧化物歧化酶(SOD)、caspase-3蛋白表达水平显著升高(P<0.05),而细胞增殖活力、丙二醛(MDA)、p-PI3K和p-AKT、Bcl-2蛋白表达水平均显著降低(P<0.05)。PI3K/AKT信号通路抑制剂的加入使各项指标差异更加显著(P<0.05)。结论过表达miR-186能够通过抑制PI3K/AKT信号通路抑制人白血病HL-60细胞的增殖并改善氧化应激水平,促进细胞凋亡。Objective The regulation of miR-186 overexpression on the proliferation,apoptosis and the phosphoinositol 3 kinase(PI3K)/protein kinase B(AKT)signaling pathway of leukemia cells was investigated.Methods Human leukemia HL-60 cells were cultured,and inverted microscope,real-time fluorescence quantitative PCR,cell counting kit 8,Hoechst 33258 staining,enzyme-linked immunosorbent assay and Western blotting analyses were used to analyze cell morphology,miR-186 expression,proliferation,apoptosis and expression level of related factors involved in oxidative stress and the PI3K/AKT pathway.Results After 24 h post-transfection,the cell growth of the miR-186 group was inhibited compared with the mimic NC group,and the relative expression level of miR-186,apoptotic cell number,superoxide dismutase factor and caspase-3 protein expression levels were significantly increased(P<0.05).Cell proliferation activity,malondialdehyde factor,p-PI3K,p-AKT and Bcl-2 protein expression levels were significantly decreased(P<0.05).There was no significant difference in PI3K and AKT protein expression levels(P>0.05).The addition of PI3K/AKT signaling pathway inhibitors made the differences of all indicators more significant(P<0.05).Conclusion The overexpression of miR-186 can inhibit the proliferation of human leukemia HL-60 cells by inhibiting the PI3K/AKT signaling pathway,improve the level of oxidative stress and promote cell apoptosis.

关 键 词：白血病HL-60细胞 miR-186 磷脂酰肌醇3激酶/蛋白激酶B 增殖 凋亡 

分 类 号：R733.7[医药卫生—肿瘤]