【摘要】 目的:探讨G蛋白偶联受体30(Gprotein-coupled receptor 30,GPR30)在人子宫内膜癌细胞株Ishikawa中的表达及亚细胞定位。方法:应用RT-PCR、蛋白质印迹法和免疫荧光技术分析Ishikawa细胞中GPR30蛋白的表达,FCM检测GPR30蛋白在细胞膜和细胞质中的阳性表达率,胶体金标记GPR30后在透射电子显微镜下观察GPR30蛋白的亚细胞定位。结果:Ishikawa细胞中有GPR30 mRNA和蛋白的表达,主要位于细胞膜和细胞质;细胞质中GPR30蛋白的阳性表达率[(20.10±0.13)%]显著高于细胞膜[(8.54±0.17)%],差异有统计学意义(P<0.01);透射电子显微镜下可见GPR30蛋白的亚细胞定位主要在细胞质的管网状网络上,可能是粗面内质网。结论:Ishikawa细胞中GPR30蛋白在细胞质中的表达高于细胞膜,其亚细胞定位主要位于粗面内质网,提示GPR30可能在子宫内膜癌中发挥重要作用。更多还原

【Abstract】 Objective:To investigate the expression and subcellular localization of G protein-coupled receptor 30(GPR30) in Ishikawa endometrial adenocarcinoma cells.Methods:The expressions of GPR30 mRNA and protein in Ishikawa cells were detected by RT-PCR,Western-blotting and immunofluorescence assay.The positive expression rate of GPR30 in the cell membrane and cytoplasm was determined by flow cytometry(FCM).The subcellular localization of GPR30 labeled with colloidal gold was observed under a transmission electron microscope.Results:The expressions of GPR30 mRNA and protein were both detectable and predominately localized in the cell membrane and cytoplasm of Ishikawa cells.The positive expression rate of GPR30 protein in the cytoplasm [(20.10±0.13)%] was significantly higher than that in the cell membrane [(8.54±0.17)%](P<0.01).The subcellular localization of GPR30 protein was mainly in the mesh-like network of cytoplasm,which may refer to rough endoplasmic reticulum.Conclusion:The expression level of GPR30 is higher in cytoplasm than in cell membrane of Ishikawa cells.GPR30 is predominantely localized in the rough endoplasmic reticulum of Ishikawa cells,which suggests an important role of GPR30 in endometrial adenocarcinoma.更多还原