MAPK/ERK通过诱导纤维蛋白A磷酸化抑制垂体瘤细胞增殖、凋亡、迁移及相关信号转导

MAPK/ERK inhibits proliferation,apoptosis,migration and related signal transduction of pituitary tumor cells by inducing fibrin A phosphorylation

目的探讨MAPK/ERK对垂体瘤细胞增殖、凋亡、迁移及相关信号转导的影响。方法通过CCK-8法确定SB203580(MAPK/ERK抑制剂)的最佳安全浓度,然后将体外培养的垂体瘤细胞分为SB203580组和对照组,通过CCK-8法检测垂体瘤细胞的增殖情况,流式细胞术检测垂体瘤细胞的细胞凋亡百分数,RT-PCR和Western blot检测垂体瘤细胞中细胞周期因子CyclinD1、CyclinE1、p21和细胞凋亡因子Bcl-2和Bax的表达情况,Western blot检测垂体瘤细胞纤维蛋白A的磷酸化水平、NF-κB、MMP-9、VEGF蛋白的表达情况,通过Transwell TM实验检测垂体瘤细胞的迁移情况。结果SB203580的最佳安全浓度是55μmol/L。SB203580处理垂体瘤细胞后,SB203580组细胞与对照组相比显著增多,CyclinD1、CyclinE1表达升高,p21、Bcl-2表达下降,Bax表达升高,纤维蛋白A的磷酸化水平下降,NF-κB、MMP-9、VEGF蛋白表达升高;流式细胞术检测显示,SB203580组的细胞凋亡数显著增加;Transwell TM实验发现,细胞迁移能力升高。结论MAPK/ERK通过促进纤维蛋白A的磷酸化,抑制垂体瘤细胞的增殖、凋亡和迁移能力,同时抑制NF-κB/MMP-9/VEGF通路的发生。

Objective To investigate the effects of MAPK/ERK on proliferation,apoptosis,migration and related signal transduction of pituitary tumor cells.Methods The optimal safe concentration of SB203580(MAPK/ERK inhibitor)was determined by CCK-8 method.Then the pituitary tumor cells cultured in vitro were divided into SB203580 group and control group,and the proliferation of pituitary tumor cells was detected by CCK-8 method;the percentage of apoptosis in pituitary tumor cells was detected by flow cytometry;RT-PCR and Western blot were used to detect the expression of cell cycle factors(CyclinD1,CyclinE1,p21)and apoptotic factors(Bcl-2 and Bax)in pituitary cells.Western blot was used to detect the phosphorylation level of fibrin A,and the expression of NF-κB,MMP-9 and VEGF protein in pituitary cells.The migration of pituitary tumor cells was detected by Transwell TM assay.Results The optimal safe concentration of SB203580 was 55μmol/L.After SB203580 treatment,compared with control group,the cells in SB203580 group were increased significantly;the expression of CyclinD1 and CyclinE1 was increased;the expression of p21 and Bcl-2 was decreased,while the expression of Bax was increased;the phosphorylation level of fibrin A was decreased,and the expression of NF-κB,MMP-9 and VEGF protein was increased.Flow cytometry showed a significant increase in the number of apoptosis in SB203580 group,and Transwell TM assay showed an increase in cell migration ability.Conclusion MAPK/ERK inhibits the proliferation,apoptosis and migration of pituitary cells by promoting the phosphorylation of fibrin A,and inhibits the occurrence of NF-κB/MMP-9/VEGF pathway.