硼替佐米对成肌细胞株C2C12的作用及其机制

发布时间：2018-06-05 12:28

本文选题：硼替佐米 + C2C12　；参考：《苏州大学》2015年硕士论文

【摘要】：目的:旨在探讨硼替佐米对小鼠成肌细胞系C2C12的生长和分化的影响并探索影响过程中参与的信号转导通路。方法:(1)利用低浓度马血清(2%)体外诱导细胞分化,吉姆萨染色后显微镜观察;(2)吉姆萨染色观察硼替佐米对C2C12细胞分化的影响;(3)MTT法检测硼替佐米对C2C12细胞增殖的影响;(4)利用流式细胞术检测硼替佐米对C2C12细胞凋亡、周期的影响;(5)利用免疫印迹法和免疫荧光法检测硼替佐米对C2C12细胞分化、周期及凋亡影响的作用机制。结果:硼替佐米能抑制多核肌管的形成并伴随着Myogenin的低表达。用不同浓度的硼替佐米处理C2C12细胞,能引起C2C12细胞增殖能力浓度依赖性的降低。Annexin V/PI染色分析显示用硼替佐米处理C2C12细胞24h后能导致细胞显著性的凋亡,Cleaved caspas-3的检测也更加证实了这一点。硼替佐米引起的细胞凋亡和p-ERK蛋白表达降低有关。细胞周期结果显示用硼替佐米孵育24h会引起G2/M的累积。结论:结果表明医用硼替佐米浓度能抑制C2C12细胞的分化,抑制C2C12细胞的增殖,诱导C2C12细胞凋亡,引起C2C12细胞G2/M期的累积。细胞水平上的研究不支持硼替佐米临床上用于骨骼肌萎缩的治疗。
[Abstract]:Aim: to investigate the effects of bortezomil on the growth and differentiation of mouse myoblast cell line C2C12 and to explore the signal transduction pathway involved in the process. Methods the cell differentiation was induced in vitro by using low concentration equine serums. The effect of bortezomil on the differentiation of C2C12 cells was observed by Giemsa staining. The effect of bortezomil on the proliferation of C2C12 cells was detected by MTT method. Flow cytometry was used to detect the apoptosis of C2C12 cells by bortezomil. The effect of bortezomib on the differentiation, cell cycle and apoptosis of C2C12 cells was studied by Western blot and immunofluorescence. Results: bortezomib inhibited the formation of polynuclear myotube and associated with the low expression of Myogenin. C2C12 cells were treated with bortezomil at different concentrations. The results of Annexin V/PI staining showed that bortezomib could induce the apoptosis of C2C12 cells after 24 hours. The results showed that the apoptosis of C2C12 cells was more obvious after treated with bortezomib for 24 hours. The results showed that the apoptosis of C2C12 cells was also confirmed by the assay of Cleaved caspas-3 after treatment with bortezomib for 24 hours. The apoptosis induced by bortezomib is related to the decrease of p-ERK protein expression. Cell cycle results showed that the accumulation of G _ 2 / M was induced by bortezomib for 24 hours. Conclusion: the results showed that the concentration of bortezomil could inhibit the differentiation of C2C12 cells, inhibit the proliferation of C2C12 cells, induce apoptosis of C2C12 cells, and induce the accumulation of G2 / M phase in C2C12 cells. Studies at the cell level do not support the clinical use of bortezomie in the treatment of skeletal muscle atrophy.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R685

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