小鼠胚胎干细胞钙敏感受体的表达和其对细胞增殖的影响

摘要

AIM:To observe the functional expression of calcium-sensing receptor (CaSR) in the mouse em-bryonic stem cells (mESCs).METHODS:The expression and distribution of CaSR were detected by Western blotting and immunofluorescence observation in 129 mouse ES-D3 cells.The intracellular concentration of free calcium ([Ca2+]i) was determined by confocal laser scanning microscopy .The cell viability was analyzed by MTT assay and flow cytometry .RE-SULTS:CaSR protein was expressed in mESCs .Extracellular calcium or neomycin significantly increased the expression of CaSR and [Ca2+]i.Neomycin increased the cell viability , up-regulated the protein expression of p-ERK2.These effects of neomycin were inhibited by NPS2390.CONCLUSION:CaSR is expressed in mESCs .The activation of CaSR is involved in the proliferation of mESCs .%目的：观察钙敏感受体（calcium-sensing receptor，CaSR）在小鼠胚胎干细胞（mouse embryonic stem cells，mESCs）的功能性表达。方法：（1）采用Western blotting 技术及免疫荧光染色分析CaSR蛋白在129小鼠ES-D3细胞系的表达；采用激光共聚焦扫描技术检测不同处理因素（包括G蛋白-PLC-IP3通路）对细胞内钙离子浓度（[ Ca2＋] i ）的影响；应用MTT法和流式细胞术分析CaSR活化对mESCs增殖的影响。结果：CaSR蛋白在mESCs有表达；增加细胞外钙或者给予CaSR激动剂新霉素均可引起[ Ca2＋] i和CaSR表达增加；CaSR活化可增加mESCs 的存活率和磷酸化ERK2蛋白的表达。 NPS2390（ CaSR抑制剂）则能减弱这种作用。结论： CaSR在mESCs有表达；CaSR的活化参与了mESCs的增殖。