miR-21与Nocodazole联合作用对小鼠成肌细胞C2C12周期的影响  

Effect of miR-21 and Nocodazole on Cell Cycle of Mouse Myoblasts C2C12

作　　者：刘会杰[1] 敖红[2] 白立景[2] 王志军 邢凯[1] 王楚端[1] 

机构地区：[1]中国农业大学动物科技学院,北京100193 [2]中国农业科学院畜牧兽医研究所,北京100193 [3]天津宁河原种猪场,天津301504

出　　处：《中国畜牧杂志》2015年第1期10-14,共5页Chinese Journal of Animal Science

基　　金：北京市现代农业产业技术创新团队(25019126)

摘　　要：观察miR-21与Nocodazole联合作用对小鼠成肌细胞C2C12周期的影响。分别以0、200、300、400、500、600 nmol/L Nocodazole处理C2C12细胞24 h,流式细胞仪检测细胞周期,间接免疫染色激光共聚焦显微镜观察细胞有丝分裂器α-微管蛋白(α-tubulin)排列。转染miR-21 mimics/NC和inhibitors/NC,24 h后,添加400 nmol/L的Nocodazole处理24 h,流式细胞仪检测细胞周期。结果表明:Nocodazole使C2C12细胞同步化的最佳浓度是400 nmol/L;过表达miR-21 mimics之后,与对照组相比,G0/G1,S期细胞百分比极显著增加,G2/M期细胞百分比极显著减少(P<0.01);添加抑制剂后,与对照组相比,添加抑制剂(inhibitors)的C2C12细胞中处于G0/G1期的细胞比例显著高于对照组,而处于G2/M期的细胞比例显著低于对照组细胞(P<0.05);正反试验结果证明,miR-21促进C2C12细胞周期进入S期。为进一步研究miR-21对C2C12细胞的作用机制奠定基础。To investigate the effect of miR-21 and Nocodazole on cell cycle of mouse myoblasts C2C12. Methods： C2C12 cells were treated with 0, 200, 300, 400, 500 and 600 nmol/L Nocodazole for 24 h respectively, the cell cycle was detected by flow cytometry. Besides, the arrangement of α-tubulin of cells was observed by confocal microscopy with indirect immunofluorescence staining. After miR-21 mimics/NC and inhibitors/NC were transfected for 24 h, the C2C12 was treated with 400 nmol/L Nocodazole for 24 h, the cell cycle was detected. Results showed that synthesized the result of FACSplus and CSLM, 400 nmol/L Nocodazole is the best for synchronization of C2C12. The effect of miR-21 on cell cycle was that miR-21 mimics vs. NC the percentage of G0/G1 and S phase in the treatment was very significantly higher than that of the control group , G2 phase was significantly lower than the control group （P〈0.01）. Inhibitors vs. NC, the percentage of Go/G1 in the treatment was significantly higher than the control group, G2 phase was significantly lower than the control group. Hence, miR-21 promotes C2C12 cell cycle into S phase. It would provide the basis for the further study of the molecular mechanism of miR-21 in the C2C12.

关 键 词：NOCODAZOLE MIR-21 C2C12 周期 小鼠 

分 类 号：S865.2[农业科学—野生动物驯养]