nmhaFGF对细胞增殖活性及信号转导的影响.docVIP

中文摘要 目的：通过对切除N端1-位氨基酸残基Grb2和MAPK-ERk1/2信号分子表达。ECV304细胞信号转导蛋白MAPK-ERK1/2研究中，我们发现nmhaFGF ECV304处理细胞30min后，胞浆MAPK-ERK1/2蛋白的表达量降低，但是在胞核中MAPK-ERK1/2其表达量更低，明显低于对照组。 4.对原代培养的人脐静脉内皮细胞的研究得出，nmhaFGF仍具有诱导人脐静脉血管内皮细胞NO生成的作用，但与haFGF组相比同等浓度下诱导NO生成的能力明显减低。 结论： nmhaFGF对NIH3T3,H22, ECV304三种细胞株增殖活性均明显降低，对NIH3T3细胞膜受体亲和力也降低，Western半定量检测结果均显示，nmhaFGF组信号蛋白ERK的表达水平明显低于haFGF组。虽然对原代培养的人脐静脉内皮细胞仍然具有一定的诱导内皮细胞产生NO的能力，但也有所降低。这些研究结果均提示nmhaFGF下调MAPK信号通路是引起nmhaFGF促增殖活性降低和诱导NO生成减少的主要原因之一。 [关键词] nmhaFGF；促增殖活性；信号转导；NO Abstract Objective:To study the mitogenesis and its mechanism of non-mitogenetic human acidic fibroblast growth factor (nmhaFGF) which lacking 27 amino acids at N-terminal，and dicuss the relationships between the mitogenesis, receptors affinity and signal transduction. The mitogenic effects of Human Acidic fibroblast growth factor (haFGF) and nmhaFGF on NIH3T3，H22，ECV304 cells were measured by MTT?and improving MTT method. The receptors affinity (Kd and IC50) and the number of binding receptors (RT) of haFGF and nmhaFGF was assessed by Radio-ligand Binding Assay of Receptors. The hepatocarcinoma cell cycle was analysised by Flow CytoMeter (FCM).The expression of the MAPK signal protein induced respectively by haFGF and nmhaFGF was detected by semi-quantitative Western-blot. The production of NO induced respectively by haFGF and nmhaFGF in culture medium was detected by Griess method respectively. Results:1. The mitogenic effect of nmhaFGF on NIH3T3 cells reduced comparable to that of haFGF. The receptors affinity of nmhaFGF also decreased, while the number of binding receptors (RT) of nmhaFGF was unchangeable. 2. The mitogenic activity and the ratio of G1 phase cells of nmhaFGF were markedly higher than that of the haFGF ,and the ratio of S phase cells were lower than that of the haFGF, but close to that of the control group.The results of Western-blot showed that the expression level of ERK of the nmhaFGF treated group on H22 was lower than