rhddADAM15蛋白抑制小鼠黑色素瘤细胞生长及其机制

Inhibitory effects and mechanism of rhddADAM15 on the growth of melanoma B16 cells

目的探讨重组人ADAM15去整合素结构域蛋白(rhddADAM15)对小鼠黑色素瘤细胞B16体外增殖、迁移、侵袭和凋亡等细胞行为的影响以及对p38丝裂原激活蛋白激酶(p38MAPK)信号通路的作用。方法 MTT法检测rhd-dADAM15对B16细胞增殖的抑制作用;划痕实验观察rhdd-ADAM15对B16细胞迁移的影响;"侵袭小室法"检测rhdd-ADAM15对B16细胞侵袭的作用;流式细胞术分析细胞周期变化;Western blot法检测rhddADAM15作用导致的p38MAPK激酶活性的变化。结果 rhddADAM15作用明显抑制B16细胞生长(IC50为13.80 mg.L-1),当浓度为7.5mg.L-1时,侵袭细胞数较对照组减少了0.55,对B16细胞迁移的抑制作用与共培养的时间呈正比,主要将B16细胞生长阻滞在G2/M期;当rhddADAM15浓度为10 mg.L-1时,p38MAPK激酶的磷酸化程度达0.80。结论 rhddADAM15对B16细胞行为具有明显的抑制作用,其机制可能与激活p38MAPK信号通路有关。

Aim To investigate the effects of rhddADAM15 on B16 cell proliferation, migration, invasion, and the eell cycles, and the involvement of p38MAPK. Methods MTF assay was employed to determine the eytotoxicity of rhddADAM15 on B16 cell growth. The wound migration assay and the Transwell chambers were used to test the effect of rhddADAM15 on B16 cell migration and invasion, respectively. The cell cy- cle and apoptosis were determined by flow cytometry, and the phosphorylation of p38 kinase in B16 cell was detected by Western blot analysis. Results rhddAD- AM15 inhibited B16 cell proliferation in a dose-de- pendent mode, and IC50 was 13.80 mg L-I. It de-creased cell migration and at the concentration of 7.5 mg ~ L-1, the number of invasive ceils was reduced by O. 55 compared with the control. It also arrested B16 cell in G2/M phase. Furthermore, phosphorylation of p38 kinase in B16 cell was detected upon treatment with rhddADAM15 （0 -10 mg· L-1 ）. Conclusion rhddADAM15 distinctively inhibits the proliferation, migration, invasion and the cell cycle of the B16 cells and the p38MAPK signal transduction pathway is in- volved in the effect of rhddADAM15 on B16 cells.