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

发布时间：2018-05-03 23:32

本文选题：miR-b + 骨肉瘤　；参考：《细胞与分子免疫学杂志》2017年10期

【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。
[Abstract]:Objective to investigate the effect and mechanism of miR-125b on aerobic glycolysis of HOS osteosarcoma cells. Methods the expression of miR-125b in HOSB normal osteoblasts and HOS osteosarcoma cells was detected by real-time quantitative PCR, the glucose uptake rate was detected by 3H-labeled 2-deoxyglucose 3H-2DG method, and the lactic acid content in supernatant was measured by quantitative lactic acid kit. To evaluate the effect of miR-125b mimic compound miR-125b-mimicson aerobic glycolysis of HOS osteosarcoma cells. Double luciferase reporter gene assay was used to determine whether hexokinase 2 or HK2 was a direct target molecule of mimics-125b, and the effect of mimics-125b on HK2 protein level was detected by Western blot assay. Results the expression level of mimics-125b in HOS osteosarcoma cells was significantly lower than that in HOSB normal osteoblasts after overexpression of miR-125b, and the glucose uptake and lactic acid production were significantly decreased. The expression of HK2 protein was significantly inhibited by aerobic glycolysis after overexpression of miR-125b in HOS cells with HK2 protein as a direct target molecule of miR-125b. Conclusion the expression of miR-125b in HOS osteosarcoma cells was low, and the aerobic glycolysis of HOS cells was inhibited by targeting the HK2 expression.
【作者单位】：赣南医学院人体解剖学教研室;赣南医学院第一附属医院骨科;赣南医学院;赣南医学院形态学实验教学中心;
【基金】：国家自然科学基金(81560039)
【分类号】：R783.1

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