摘要
为探讨miR-200c对人乳腺癌MCF-7 (Michigan cancer foundation-7)细胞糖代谢水平的影响,本研究使用miR-200c mimics转染MCF-7细胞,通过定量即时聚合酶链锁反应(quantitative real time polymerase chain reaction, qRT-PCR)检测各组细胞miR-200c的表达水平;利用细胞计数盒(cell counting kit-8, CCK-8)检测miR-200c mimics转染对MCF-7细胞增殖的影响;通过葡萄糖试剂盒检测各组细胞葡萄糖的消耗,乳酸试剂盒检测各组细胞乳酸的释放量;通过蛋白质印迹法(Western blotting)检测各组细胞糖代谢相关酶己糖激酶(hexokinase 2, HK2)以及肌肉丙酮酸激酶同工酶2 (pyruvate kinase isozyme type M2, PKM2)蛋白表达水平。与miR-NC组相比,miR-200c mimics转染明显上调MCF-7细胞miR-200c m RNA水平;且miR-NC组和MCF-7组miR-200c mRNA水平没有明显差异;细胞计数盒(cell counting kit-8, CCK-8)检测结果显示,与miR-NC组和MCF-7组相比,miR-200c mimics转染不仅明显降低乳腺癌MCF-7细胞的细胞活力,而且显著减少乳腺癌MCF-7细胞葡萄糖的消耗;此外,Western blotting结果显示,miR-200c显著下调MCF-7细胞糖代谢相关酶HK2和PKM2的表达。综上结果表明,miR-200c会抑制人乳腺癌MCF-7细胞糖代谢。本研究成果为乳腺癌防治提供一定的参考价值。
To investigate the impact of miR-200 c on glucose metabolism in breast cancer MCF-7(michigan cancer foundation-7)cell.MCF-7 cells were transfected with miR-200 c mimics,and,the m RNA level of miR-200 c in MCF-7 cell was measured by quantitative real time polymerase chain reaction(qRT-PCR).The effect of miR-200 c on MCF-7 cell proliferation was detected by cell counting kit(CCK-8).Glucose consumption was measured by glucose assay kits and the lactate production was measured by lactate assay kits.The expressions of glucose metabolism-related proteins,including hexokinase 2(HK2)as well as pyruvate kinase isozyme type M2(PKM2),were assayed by Western blotting.Compared to miR-NC group,miR-200 c mimics transfection dramatically increased the mRNA expression of miR-200 c in MCF-7 cell.And the difference of miR-200 c mRNA expression between miR-NC group and MCF-7 group had no significance.Data from CCK-8 showed that compared with the miR-NC group and the MCF-7 group,miR-200 c mimics transfection not only significantly reduced the cell viability of breast cancer MCF-7 cells,but also significantly reduced the glucose consumption of breast cancer MCF-7 cells.Furthermore,data from Western blotting exhibited that miR-200 c dramatically decreased the expression of the enzymes involved in glucose metabolism in cells,HK2 as well as PKM2 in MCF-7 cell.In summary,miR-200 c could inhibits glucose metabolism in human breast cancer MCF-7 cells.This research provide a certain reference value for breast cancer prevention and treatment.
作者
陈华波
张崇
殷焦
肖娟
杨林
Chen Huabo;Zhang Chong;Yin Jiao;Xiao Juan;Yang Lin(HubeiUniversity ofArts and Science,Xiangyang 441053;Xiangyang First People'sHospital,Xiangyang,441053)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2019年第6期2874-2878,共5页
Genomics and Applied Biology
基金
2018年地方高校国家级大学生创新创业训练计划项目(201810519002)
湖北省教育厅科学技术研究项目(D20172603)共同资助
