BMP, Mitf, and RET on melanocytes and melanoma cell

Western blotting结果显示，Ret蛋白在NCCmelb4M5和Mel-Ret细胞中表达，而在NCCmelb4和NCCmelan5细胞中未表达。免疫染色还显示，NCCmelb4M5和Mel-Ret细胞Ret表达阳性，而NCCmelb4和NCCmelan5细胞Ret表达阴性。因此，Ret蛋白在大多数未成熟的黑素母细胞中表达，而黑素细胞Ret表达阴性。将BMP-4加入培养液中，以不同浓度孵育3天。10 ng/ml BMP-4孵育3天后，Ret蛋白表达降低，20 ng/ml BMP-4孵育后Ret蛋白表达完全消失。BMP-4诱导的NCCmelb4M5细胞中Ret蛋白表达呈剂量依赖性降低。基于这些发现，Ret和BMP-4的联合作用可能为理解黑素细胞发育中的作用和工作机制提供重要线索。

Western blotting revealed expression of the Ret protein in NCCmelb4M5 and in Mel-Ret cells, but in contrast, there was no expression of the Ret protein in NCCmelb4 or NCCmelan5 cells. Immunostaining also revealed that NCCmelb4M5 and Mel-Ret cells are positive for Ret, but NCCmelb4 and NCCmelan5 cells are negative for Ret. Thus, Ret protein is expressed in most immature melanoblasts, while melanocytes are negative for Ret. BMP-4 was added to the medium and incubated for 3 days at varying concentrations. After incubation with 10 ng/ml BMP-4 for 3 days, Ret protein expression was decreased, and disappeared completely when BMP-4 was added at 20 ng/ml. Ret protein expression in NCCmelb4M5 cells showed a dose-dependent decrease in expression elicited by BMP-4. Based on these findings, the combined effects of Ret and BMP-4 might provide important clues towards understanding the roles and working mechanisms in melanocyte development.