摘要
目的研究miR-451对肝癌细胞增殖、糖酵解及相关基因表达的影响。方法实验分为普通组、增强组和抑制组,增强组和抑制组肝癌MHCC97细胞分别转染miR-451的模拟物mimics和阻遏物inhibitors,普通组常规培养。采用RT-PCR法检测3组肝癌细胞中miR-451和AMPK/mT OR、Ki-67、血管内皮生长因子(VEGF)、基质金属蛋白酶9(MMP-9)、胰岛素样生长因子(IGF-1)表达情况,噻唑蓝(MTT)法检测3组肝癌细胞增殖情况,通过检测3组细胞葡萄糖摄取和乳酸分泌情况评估肝癌细胞糖酵解情况。结果增强组肝癌细胞中miR-451表达量明显高于普通组(P<0.05),抑制组肝癌细胞中miR-451表达量明显低于普通组(P<0.05)。MTT结果显示转染后各时间点增强组吸光度(OD)明显低于普通组和抑制组(P均<0.05)。转染后增强组肝癌细胞mT OR、Ki-67、MMP-9、VEGF、IGF-1mR NA表达量和葡萄糖消耗值、乳酸分泌值均明显低于普通组和抑制组(P均<0.05),而AMPK mR NA表达量明显高于普通组和抑制组(P均<0.05)。结论 miR-451参与调控肝癌细胞增殖及糖酵解,其调控机制可能通过影响AMPK/mT OR信号通路,进而影响细胞中Ki-67、MMP-9、VEGF、IGF-1的表达而实现。
Objective It is to investigate the effect of mi R-451 on the capacity of proliferation,glycolysis and related gene expression of hepatoma cells. Methods There was common group,enhanced group and suppressing group. The hepatoma MHCC97 cells in the enhanced group and inhibitory group were transfected with the mimics and inhibitors of mi R-451,respectively; and with normal culture in common group. The expression of mi R-451,and AMPK/m TOR,Ki-67,vascular endothelial growth factor( VEGF),matrix metalloproteinase 9( MMP-9),insulin like growth factor( IGF-1) in hepatoma cells were detected by RT-PCR in the 3 groups; the capacity of proliferation in hepatoma cells was detected by thiazolium( MTT) method in the 3 groups; and glycolysis in hepatoma cells was evaluated by detecting glucose uptake and lactate secretion in the 3 groups. Results The expression of mi R-451 in hepatoma cells of the enhanced group was significantly higher than that of the normal group( P < 0. 05). The expression of mi R-451 in hepatoma cells of the inhibition group was significantly lower than that of the normal group( P < 0. 05). The results of MTT showed that the absorbance of the enhanced group at each time point after transfection was significantly lower than that of the normal group and the inhibition group( all P < 0. 05). After transfection,the expression level of m TOR,Ki-67,MMP-9,VEGF and IGF-1 m RNA and the value of glucose consumption and lactate secretion in the hepatoma cells were significantly lower than those in the normal group and the inhibition group( all P < 0. 05),but the expression of AMPK m RNA was significantly higher than that of the common group and the inhibition group( P < 0. 05). Conclusion mi R-451 is involved in regulating the proliferation and glycolysis of hepatoma cells,the regulatory mechanism may be affect the expression of Ki-67,MMP-9,VEGF and IGF-1 by influence the AMPK/m TOR signaling pathway in the cells.
出处
《现代中西医结合杂志》
CAS
2018年第9期934-937,共4页
Modern Journal of Integrated Traditional Chinese and Western Medicine
作者简介
周艳华,女,主治医师,硕士,主要从事恶性肿瘤的临床诊治与基础研究工作.;[通信作者]戴广海,E-mail：daigh301@vip.sina.com
