miR-138-5p靶向HIF-1α影响人白血病K562细胞增殖和侵袭转移  

Effect of miR-138-5p targeting HIF-1αon proliferation and invasion and metastasis of human leukemia K562 cells

作　　者：郭涛 纪冬梅 李艳平 卢阳 弋莉 王晓红 GUO Tao;JI Dong-mei;LI Yan-ping;LU Yang;YI Li;WANG Xiao-hong(Department of Hematology,Chaoyang Central Hospital,Chaoyang Liaoning 122000,China)

机构地区：[1]朝阳市中心医院血液内科,辽宁朝阳122000

出　　处：《局解手术学杂志》2020年第4期272-277,共6页Journal of Regional Anatomy and Operative Surgery

基　　金：辽宁省自然科学基金(2015020372)。

摘　　要：目的探索miR-138-5p与缺氧诱导因子-1α(HIF-1α)对人白血病K562细胞增殖和侵袭转移的影响。方法以体外培养的人白血病K562细胞为研究对象,将其分为K562组、miR-138 mimic组、pc-HIF-1α组和mimic+pc-HIF-1α组。荧光素酶报告实验确认miR-138-5p与HIF-1α的靶向关系;qRT-PCR法检测miR-138-5p和HIF-1α的mRNA水平;Western blot检测HIF-1α的蛋白水平;CCK-8检测细胞存活率;Transwell检测细胞侵袭能力;划痕实验检测细胞迁移能力;Western blot检测增殖和上皮间质转化(EMT)相关蛋白表达水平。结果miR-138-5p能靶向HIF-1α,与K562组相比,miR-138 mimic组HIF-1α的mRNA和蛋白水平均显著较低(P<0.01),而pc-HIF-1α组HIF-1α的蛋白水平则显著较高(P<0.01);同时,与pc-HIF-1α组相比,mimic+pc-HIF-1α组HIF-1α的蛋白表达显著较低(P<0.01)。与K562组相比,miR-138 mimic组细胞增殖倍数、细胞迁移和侵袭能力均显著较低(P<0.01),pc-HIF-1α组则显著较高(P<0.01);与pc-HIF-1α组相比,mimic+pc-HIF-1α组细胞增殖倍数、细胞迁移和侵袭能力均显著较低(P<0.01)。同时,mimic+pc-HIF-1α能部分逆转pc-HIF-1α对肿瘤细胞增殖和EMT能力的上调作用(P<0.01)。结论miR-138-5p可通过靶向HIF-1α,进而降低人白血病K652细胞增殖和侵袭转移能力。Objective To explore the effects of miR-138-5p and hypoxia-inducible factor-1-alpha(HIF-1α)on the proliferation,invasion and metastasis of human leukemia K562 cells.Methods K562 cells cultured in vitro were randomly divided into the K562 group,the miR-138 mimic group,the pc-HIF-1αgroup,and the mimic+pc-HIF-1αgroup.The targeted relationship of miR-138-5p and HIF-1αwas affirmed by luciferase assay.The expression of miR-138-5p and mRNA levels of HIF-1αwere measured by qRT-PCR.The protein levels of HIF-1αwere detected by Western blot.Cell proliferation was tested by CCK-8.Cell invasion was detected by Transwell.Cell migration was tested by scratch assay.And Western blot was used to detect the expression levels of proliferative related proteins and epithelial mesenchymal transformation(EMT)related proteins.Results miR-138-5p could target HIF-1α.The mRNA and protein levels of HIF-1αin the miR-138 mimic group were lower than those in the K562 group(P<0.01).The protein levels of HIF-1αin the pc-HIF-1αgroup were higher than those in the K562 group(P<0.01).Compared with the pc-HIF-1αgroup,the protein levels of HIF-1αin the mimic+pc-HIF-1αgroup were significantly lower(P<0.01).Compared with the K562 group,the proliferation,migration and invasion of the cells in the miR-138 mimic group were significantly lower(P<0.01),while those in the pc-HIF-1αgroup were significantly higher(P<0.01).Compared with the pc-HIF-1αgroup,the proliferation,migration and invasion of the cells in the mimic+pc-HIF-1αgroup were lower(P<0.01).At the same time,mimic+pc-HIF-1αcould partially reverse the up-regulation of pc-HIF-1αon proliferation and EMT ability of K562 cells(P<0.01).Conclusion miR-138-5p can reduce the proliferation and invasion and metastasis of human leukemia K652 cells by targeting HIF-1α.

关 键 词：白血病 慢性粒细胞白血病 miR-138-5p HIF-1Α 增殖 上皮间质化 

分 类 号：R733.72[医药卫生—肿瘤]