miR-125b通过下调己糖激酶2的表达抑制HOS骨肉瘤细胞有氧糖酵解  

miR-125b suppresses the aerobic glycolysis of osteosarcoma HOS cells by downregulating the expression of hexokinase-2

作　　者：吴玉娟[1] 何华宾[2] 吴冰[1] 温军 郭哲 罗煜 曹镐禄[4] WU Yujuan;HE Huabin;WU Bing;WEN Jun;GUO Zhe;LUO Yu;CAO Gaolu(Department of Human Anatomy, Gannan Medical University;Department of Orthopaedics, First Affiliated Hospital, Gannan Medical University;Gannan Medical University;Experimental Teaching Center of Morphology, Gannan Medical University, Ganzhou 341000, China)

机构地区：[1]赣南医学院人体解剖学教研室,江西赣州341000 [2]赣南医学院第一附属医院骨科,江西赣州341000 [3]赣南医学院,江西赣州341000 [4]赣南医学院形态学实验教学中心,江西赣州341000

出　　处：《细胞与分子免疫学杂志》2017年第10期1365-1370,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81560039)

摘　　要：目的探讨miR-125b对HOS骨肉瘤细胞有氧糖酵解的影响和机制。方法采用实时定量PCR检测HOSB正常成骨细胞和HOS骨肉瘤细胞中miR-125b的表达水平;采用3H标记的2-脱氧葡萄糖(3H-2DG)法检测细胞葡萄糖摄取率、乳酸定量试剂盒检测细胞上清中乳酸含量,以评价miR-125b模拟物(miR-125b-mimics)对HOS骨肉瘤细胞有氧糖酵解的影响。采用双荧光素酶报告基因实验明确己糖激酶2(HK2)是否为mimics-125b的直接靶分子;采用Western blot法检测mimics-125b对HK2蛋白水平的影响。结果 mimics-125b在HOS骨肉瘤细胞中表达水平显著低于HOSB正常成骨细胞;HOS细胞过表达miR-125b后,糖摄取率和乳酸生成明显减少,有氧糖酵解受到显著抑制;HK2蛋白为miR-125b的直接靶分子;HOS细胞过表达miR-125b后,HK2蛋白表达受到抑制。结论 miR-125b在HOS骨肉瘤细胞中低表达,并通过靶向抑制HK2的表达抑制HOS细胞有氧糖酵解。Objective To investigate the effect of miR-125 b on the aerobic glycolysis of osteosarcoma HOS cells and its underlying mechanism. Methods Real-time quantitative PCR was performed to detect the expression of miR-125 b in HOSB normal human osteoblast cells and HOS osteosarcoma cells. The glucose uptake rate was assessed with3 H-2 deoxyglucose（3 H-2 DG） and lactate production was tested with the kits to observe the effect of miR-125 b-mimics on the aerobic glycolysis of osteosarcoma HOS cells. Hexokinase-2（ HK2） protein was detected by Western blot analysis. Dual luciferase reporter gene assay was used to determine whether HK2 was the direct target of miR-125 b. Results Compared with HOSB normal human osteoblast cells,the expression of miR-125 b was significantly lower in HOS cells. The glucose uptake and lactate production were downregulated in HOS cells transfected with miR-125 b-mimics. Aerobic glycolysis of HOS cells was markedly inhibited. Protein level of HK2 was significantly inhibited in HOS cells transfected with miR-125 b-mimics. Luciferase assay indicated that HK2 was the direct target of miR-125 b. Conclusion miR-125 b is down-regulated in HOS cells,and it inhibits the aerobic glycolysis of HOS cells by directly regulating the expression of HK2.

关 键 词：miR-125b 骨肉瘤 有氧糖酵解 己糖激酶2(HK2) 

分 类 号：R783.1[医药卫生—口腔医学] R738.1[医药卫生—临床医学]