采用MDA-MB-231细胞膜进行表皮生长因子受体活性的检测

Activity evaluation of epidermal growth factor receptor by MDA-MB-231 plasma membrane

目的 探讨能否直接利用肿瘤细胞膜进行表皮生长因子受体 (Epidermalgrowthfactorreceptor,EGFR)催化活性检测。方法 首先筛选出EGFR基因表达水平相对较高的细胞株MDA MB 2 31,通过差速离心制备细胞膜 ,采用Westernblotting检测EGFR催化磷酸化的程度。结果 底物被磷酸化 ,加入特异性拮抗剂AG14 78后 ,磷酸化被抑制。结论 利用肿瘤细胞膜检测EGFR活性的设想是成立的 ,并且其方法简便、经济。

Objective To evaluate the epidermal growth factor receptor (EGFR) activity using plasma membrane of tumor cells. Methods The tumor cell strain MDA-MB-231 that has high EGFR gene expression level was selected, then MDA-MB-231 plasma membrane was obtained by differential centrifugation. EGFR catalytic activity was assayed by Western blotting. Results The substrate was phosphorylated and the phosphorylation was blocked after addition of the special inhibitor-AG1478. Conclusion EGFR activity in MDA-MB-231 plasma membrane could be determined by the method described above