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C2C12-GLUT4HA骨骼肌细胞株的筛选 被引量：2

Screening of C2C12-GLUT4HA Skeletal Muscle Cell Line
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摘要 目的筛选过表达带有HA表位的葡萄糖转运子4(GLUT4HA)且具有收缩能力的C2C12骨骼肌细胞株。方法建立稳定过表达GLUT4HA的C2C12骨骼肌细胞株,从中挑选出23个克隆。用光镜观察各个克隆分化为多核肌管的能力和响应电脉冲刺激(EPS)的收缩能力,并用免疫印迹法检测各克隆中GLUT4HA蛋白的表达水平,筛选出分化良好、响应EPS收缩且高表达GLUT4HA的细胞株。将各细胞株进一步分为胰岛素组及其对照组、EPS组及其对照组,用酶联免疫吸附(ELISA)法测定各组细胞膜上GLUT4HA的含量(转位)。结果在选出的23个克隆中,克隆17的GLUT4HA表达量较高,分化和收缩能力良好,且在胰岛素和EPS后GLUT4转位显著升高。结论本研究建立了过表达GLUT4HA的具有收缩能力的C2C12-GLUT4HA骨骼肌细胞株,可用于研究胰岛素调节GLUT4转位和收缩调节GLUT4转位的机制。 Objective To select a GLUT4HA-overexpressing C2C12 cell line with contractile activity.Methods C2C 12-GLUT4HA cell line stably overexpressed GLUT4HA was created, and 23 of these clones were selected. The differen- tiation and contraction ability, as well as the expression level of GLUT4HA protein were detected. The clones with good myotubes, contraction ability and high GLUT4HA protein level were divided into insulin group and its control group, electrical pulse stimulation （EPS） group and its control group. The amount of GLUT4HA on the cell surface was measured by enzyme-linked immunosorbent assay （ELISA） after stimulation by insulin and EPS.Results Among the selected 23 clones, clone 17 showed higher GLUT4HA protein level, good myotubes and contraction ability. In addition, GLUT4HA in clone 17 myotubes was translocated to cell surface under insulin and EPS stimulation. Conclusion A contractile C2C12 muscle cell line stably overexpressed GLUT4HA was created. This cell line will be useful to study the mechanism of insulin- and contraction stimuli-regulated GLUT4 traffic.
作者 仇利红 高静 牛文彦
机构地区 天津医科大学免疫教研室 天津中医药大学第一附属医院检验科
出处 《天津医药》 CAS 北大核心 2013年第6期580-582,共3页 Tianjin Medical Journal
基金 国家自然科学基金资助项目(项目编号:81170740) 国家自然科学基金委员会国际合作与交流项目(项目编号:81161120545)
关键词 肌骨骼 葡萄糖转运体型 细胞培养的 胰岛素 肌收缩 电脉冲刺激 muscle, skeletal glucose transporter type 4 cells, cultured insulin muscle contraction electrical pulsestimulation
分类号 R735.2 [医药卫生—肿瘤]
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参考文献10

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二级参考文献7

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共引文献1

同被引文献41

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天津医药
2013年 第6期

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