本文選題：miR-b + 骨肉瘤　； 參考：《細(xì)胞與分子免疫學(xué)雜志》2017年10期

【摘要】：目的探討miR-125b對(duì)HOS骨肉瘤細(xì)胞有氧糖酵解的影響和機(jī)制。方法采用實(shí)時(shí)定量PCR檢測(cè)HOSB正常成骨細(xì)胞和HOS骨肉瘤細(xì)胞中miR-125b的表達(dá)水平;采用3H標(biāo)記的2-脫氧葡萄糖(3H-2DG)法檢測(cè)細(xì)胞葡萄糖攝取率、乳酸定量試劑盒檢測(cè)細(xì)胞上清中乳酸含量,以評(píng)價(jià)miR-125b模擬物(miR-125b-mimics)對(duì)HOS骨肉瘤細(xì)胞有氧糖酵解的影響。采用雙熒光素酶報(bào)告基因?qū)嶒?yàn)明確己糖激酶2(HK2)是否為mimics-125b的直接靶分子;采用Western blot法檢測(cè)mimics-125b對(duì)HK2蛋白水平的影響。結(jié)果 mimics-125b在HOS骨肉瘤細(xì)胞中表達(dá)水平顯著低于HOSB正常成骨細(xì)胞;HOS細(xì)胞過表達(dá)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(xì)胞過表達(dá)miR-125b后,HK2蛋白表達(dá)受到抑制。結(jié)論 miR-125b在HOS骨肉瘤細(xì)胞中低表達(dá),并通過靶向抑制HK2的表達(dá)抑制HOS細(xì)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(xué)院人體解剖學(xué)教研室;贛南醫(yī)學(xué)院第一附屬醫(yī)院骨科;贛南醫(yī)學(xué)院;贛南醫(yī)學(xué)院形態(tài)學(xué)實(shí)驗(yàn)教學(xué)中心;
【基金】：國(guó)家自然科學(xué)基金(81560039)
【分類號(hào)】：R783.1

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