重组人ADAM15去整合素区域蛋白抑制HeLa细胞的增殖及迁移

Recombinant Human Disintegrin Domain of ADAM15 Inhibits the Proliferation and Migration of HeLa Cells

摘要:

观察重组人ADAM15去整合素区域蛋白(记做rhddADAM15)干预HeLa细胞在体内外的增殖及迁移效应, 初步鉴定了其作用的相关通路, 为研究其作用机制提供基础。利用SRB法及划痕法测得rhddADAM15对HeLa体外的增殖及迁移均有明显的抑制作用, IC50分别为2.44、1.60μmol/L, 当其浓度低于8 μmol/L时, 细胞增殖及迁移抑制率与蛋白质浓度呈剂量依赖关系; 利用瞬时转染法在HeLa细胞内过表达rhddADAM15, 致使HeLa细胞的迁移能力下降, 细胞周期出现S期阻滞, 阻滞率约为5%; SRB法表明, MAPK通路激动剂EGF可以逆转rhddADAM抑制HeLa细胞增殖的效应; 利用斑马鱼肿瘤移植模型, 观察得到rhddADAM15对斑马鱼体内HeLa细胞生长及转移均有明显的抑制作用。以上结果表明, rhddADAM15可抑制HeLa细胞体内外的增殖及迁移, 这种抑制作用与MAPK信号通路密切相关, 作用靶点在细胞外及细胞内均有分布。

Abstract:

The anti-proliferation and anti-migration activities of recombinant human disintegrin domain of ADAM15(rhddADAM15) were evaluated in vitro and in vivo and the related pathway was determined in this study. rhddADAM15 inhibited the proliferation and migration of human cervical cancer HeLa cells with the IC50 of 2.44 and 1.60 μmol/L respectively evaluated by SRB assay and wound healing assay. Over-expression of disintegrin domain of ADAM15 in HeLa cells by transient transfection induced the migration inhibition and partial S phase arrest. In addition, the inhibition effect of proliferation of HeLa cells aroused by rhddADAM15 was reversed by EGF, which is the agonist of MAPK signal pathways. Moreover, rhddADAM15 inhibited the growth and metastasis of HeLa cells xenograft in zebrafish. All the results showed that rhddADAM15 inhibited the proliferation and migration of HeLa cells by MAPK signal pathway and the target may exist both on the cell surface and in the cytoplasm.