本文選題：miR-b + 骨肉瘤　； 參考：《細胞與分子免疫學雜志》2017年10期

【摘要】：目的探討miR-125b對HOS骨肉瘤細胞有氧糖酵解的影響和機制。方法采用實時定量PCR檢測HOSB正常成骨細胞和HOS骨肉瘤細胞中miR-125b的表達水平;采用3H標記的2-脫氧葡萄糖(3H-2DG)法檢測細胞葡萄糖攝取率、乳酸定量試劑盒檢測細胞上清中乳酸含量,以評價miR-125b模擬物(miR-125b-mimics)對HOS骨肉瘤細胞有氧糖酵解的影響。采用雙熒光素酶報告基因?qū)嶒灻鞔_己糖激酶2(HK2)是否為mimics-125b的直接靶分子;采用Western blot法檢測mimics-125b對HK2蛋白水平的影響。結(jié)果 mimics-125b在HOS骨肉瘤細胞中表達水平顯著低于HOSB正常成骨細胞;HOS細胞過表達miR-125b后,糖攝取率和乳酸生成明顯減少,有氧糖酵解受到顯著抑制;HK2蛋白為miR-125b的直接靶分子;HOS細胞過表達miR-125b后,HK2蛋白表達受到抑制。結(jié)論 miR-125b在HOS骨肉瘤細胞中低表達,并通過靶向抑制HK2的表達抑制HOS細胞有氧糖酵解。
[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.
【作者單位】： 贛南醫(yī)學院人體解剖學教研室;贛南醫(yī)學院第一附屬醫(yī)院骨科;贛南醫(yī)學院;贛南醫(yī)學院形態(tài)學實驗教學中心;
【基金】：國家自然科學基金(81560039)
【分類號】：R783.1

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