多胺对PC12细胞生长和信号转导的影响

发布时间：2018-10-30 20:25

【摘要】：【目的】:研究多胺(精脒)及多胺合成抑制剂DFMO一定浓度对PC12细胞生长特性的影响,同时探讨在其作用下PC12细胞MAPKs信号转导通路活化表达的特点及其相互之间的关系。【方法】:取对数生长期嗜铬细胞瘤细胞株PC12细胞,按照作用剂的不同分为两组,即SPD组(分别含SPD1.25,2.5,5,10 mmol/L)和DFMO组(分别含DFMO 2,4,6,8 mmol/L),分别培养4天,每天观察PC12细胞生长情况。光镜显微镜下观察细胞形态学变化;台盼蓝染色法绘制细胞生长曲线;用四氮唑盐(MTT)比色试验检测细胞的生长或抑制;流式细胞仪检测细胞周期分布。同时用western-blot法检测ERK1/2和p38蛋白磷酸化水平,观察多胺对PC12细胞生长信号转导途径的影响。【结果】:光镜显微镜下观察到:5,10 mmol/L SPD作用PC12细胞4天后,细胞数较对照组明显增多,胞体更加粗大,细胞突起更加细长,分支相连,1.25,2.5 mmol/L SPD作用PC12细胞4天后,与对照组相比无明显变化;4,6 mmol/L DFMO作用PC12细胞4天后,大部分细胞皱缩变圆,细胞突起消失,出现大量漂浮细胞,2,8 mmol/L DFMO作用PC12细胞4天后与对照组相比大部分细胞变圆,细胞突起变短。台盼蓝染色法绘制细胞生长曲线可见SPD组活细胞数明显增多,DFMO组细胞数减少,而DFMO组加入SPD后与对照组相比没有明显变化。MTT
[Abstract]:[aim]: to study the effects of polyamine (spermidine) and polyamine synthesis inhibitor (DFMO) on the growth characteristics of PC12 cells, and to explore the characteristics of the activation and expression of MAPKs signal transduction pathway in PC12 cells and their relationship with each other. [methods]: the logarithmic pheochromocytoma cell line PC12 cells were divided into two groups according to the different agents: SPD group (containing SPD1.25,2.5,5,10 mmol/L) and DFMO group (containing DFMO 2 + 4 + 6 + 8 mmol/L) for 4 days, respectively. The growth of PC12 cells was observed every day. Morphological changes were observed under light microscope; cell growth curve was drawn by trypan blue staining; cell growth or inhibition was detected by tetrazolium (MTT) assay; cell cycle distribution was detected by flow cytometry. The phosphorylation levels of ERK1/2 and p38 proteins were detected by western-blot assay, and the effects of polyamines on the growth signal transduction pathway of PC12 cells were observed. [results]: it was observed under the light microscope that the number of PC12 cells increased significantly, the cell body was thicker, the cell processes were more slender, and the branches were connected with the cells treated with 5 mmol/L SPD for 4 days, compared with the control group. PC12 cells treated with 1.25 mmol/L SPD for 4 days showed no significant change compared with the control group. After 4 days of treatment with 4 mmol/L DFMO, most of PC12 cells became round, the cell processes disappeared, and a large number of floating cells appeared. After 4 days of treatment with 2 ~ 8 mmol/L DFMO, most of the cells became round and the cell processes became shorter than those in the control group. Trypan blue staining showed that the number of living cells in SPD group was significantly increased, but that in DFMO group was decreased, but that in DFMO group was not significantly different from that in control group.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R341

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