过表达miR-186减弱EMT途径抑制白血病细胞增殖、迁移及糖酵解  

Overexpression of miR-186 inhibits cell proliferation,migration and glycolysis of leukemia cells by inhibiting epithelial-mesenchymal transition

作　　者：冯永笑[1] 张丑丑[1] 董艳琴[1] 赵晓亮 FENG Yongxiao;ZHANG Chouchou;DONG Yanqin(Department of Integrated Traditional Chinese and Western Medicine,Gansu Provincial Cancer Hospital,Gansu,Lanzhou 730050,China)

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

出　　处：《河北医药》2023年第24期3703-3707,共5页Hebei Medical Journal

基　　金：甘肃省中医药管理局科研课题(编号:GZK-2014-78)。

摘　　要：目的探究过表达miR-186对白血病细胞增殖、迁移、糖酵解和上皮间质转化(EMT)的影响。方法体外培养人白血病HL-60细胞,然后转染mimic NC和miR-186 mimic至HL-60细胞,分别设为mimic NC组(阴性对照组)和miR-186过表达组,非转染的细胞设为对照组。miR-186表达水平采用实时荧光定量PCR(RT-qPCR)法测定;细胞形态用倒置显微镜观察;细胞增殖抑制率采用活细胞计数(CCK-8)法测定;细胞迁移率采用Transwell小室法测定;乳酸生成量和葡萄糖消耗水平分别采用萄糖检测试剂盒和乳酸检测试剂盒测定;EMT相关因子[E-钙粘蛋白(E-cadherin)、N-钙粘蛋白(N-cadherin)和波形蛋白(Vimentin)]表达水平使用RT-qPCR和蛋白免疫印迹(WB)法测定。结果转染24 h后,与对照组和mimic NC组相比,miR-186过表达组miR-186相对表达量、细胞增殖抑制率及E-cadherin mRNA和蛋白表达量显著增加(P<0.05),而细胞迁移率、葡萄糖消耗水平、乳酸生成量及N-cadherin、Vimentin mRNA和蛋白表达水平显著减少(P<0.05);对照组和mimic NC组比较差异无统计学意义(P>0.05)。结论过表达miR-186能够通过减弱EMT进程抑制人白血病HL-60细胞的增殖、迁移和糖酵解。Objective To explore the effect of overexpressed miR-186 on the proliferation,migration,glycolysis and epithelial-mesenchymal transition(EMT)of leukemia cells.Methods The human leukemia cell line HL-60 was cultured in vitro,and transfected with mimic negative control(NC group)and miR-186 mimic(overexpression group).Cells without transfection were regarded as the control group.The expression level of miR-186 was detected by real-time quantitative PCR(RT-qPCR).Cell morphology was observed using an inverted microscope.Cell counting kit-8(CCK-8)and Transwell assay were performed to detect cell proliferation and migration,respectively.Lactate production and glucose consumption were measured by glucose detection kit and lactate detection kit,respectively.Expression levels of EMT-related factors(E-cadherin,N-cadherin,and Vimentin)were determined using RT-qPCR and Western blotting(WB).Results After transfection for 24 h,overexpression of miR-186 significantly upregulated miR-186,and increased the proliferative inhibition rate,and mRNA and protein levels of E-cadherin,which significantly inhibited migratory rate,glucose consumption,lactate production,and mRNA and protein levels of N-cadherin and Vimentin(P<0.05).The above indexes were comparable between HL-60 cells treated with blank control and those transfected with NC(P>0.05).Conclusion Overexpression of miR186 inhibits proliferation,migration and glycolysis of human leukemia HL-60 cells by attenuating EMT process.

关 键 词：白血病 miR-186 增殖 迁移 上皮间质转化 糖酵解 

分 类 号：R733.7[医药卫生—肿瘤]