钙敏感受体逆转子宫内膜癌细胞上皮-间充质转化

目的:探讨钙敏感受体(calcium-sensing receptor,CaSR)在缺氧环境下对子宫内膜癌细胞发生上皮-间充质转化(epithelial mesenchymal transitions,EMT)的影响。方法:将子宫内膜癌Ishikawa细胞分别转染相应腺病毒,并分为空白组、空载组、沉默CaSR组和过表达CaSR组。将各组细胞进行缺氧后细胞免疫荧光(immunofluorescence,IF)法观察细胞骨架的变化;蛋白质印迹法及IF法检测钙黏附蛋白E(E-cadherin)、波形蛋白(Vimentin)、CaSR蛋白表达变化;荧光染料Fura-2/AM示踪监测细胞内钙离子的浓度变化;Transwell迁移试验检测细胞侵袭迁移能力变化。结果:缺氧后,子宫内膜癌细胞的细胞骨架呈梭形样改变;同时E-cadherin表达降低,Vimentin和CaSR表达增加;子宫内膜癌细胞转染沉默及过表达CaSR腺病毒后细胞内钙离子内流分别减少及增加,而与Vimentin表达和细胞侵袭迁移能力呈负相关。结论:缺氧后子宫内膜癌细胞发生EMT及CaSR表达增加,而CaSR的表达增加抑制EMT发生,其机制可能与调控细胞钙离子有关。

Calcium-sensitive receptors reverse epithelial-mesenchymal transformation of endometrial cancer cells

Objective: To investigate the effect of calcium sensitive receptor(CaSR) on epithelial mesenchymal transformation(EMT) of endometrial cancer cells in hypoxic environment. Methods: The Ishikawa cells of endometrial carcinoma were transfected with corresponding adenovirus respectively to form a blank group, a noload group, a silent CaSR group and an overexpressed CaSR group. Th e change of cytoskeleton in different groups was observed by immunofluorescence assay aft er hypoxia. Expression of E-cadherin, Vimentin and CaSR were detected by Western bloting and immunofluorescence. Fluorescence dye fura-2/AM tracer was used to monitor the change of intracellular calcium ion concentration. Transwell assay was used to detect change in the ability of cell invasion and migration. Results: After hypoxia, the cytoskeleton of endometrial cancer cells changed like a spindle. Meanwhile, the expression of E-cadherin decreased, while the expression of Vimentin and CaSR increased. Endometrial cancer cells transfected with silencing and overexpressed CaSR adenovirus reduced and increased intracellular calcium flow respectively, which was negatively correlated with the expression of Vimentin and the ability of cell invasion and migration. Conclusion: The expression of EMT and CaSR in endometrial cancer cells increases aft er hypoxia, and the increased expression of CaSR inhibites the occurrence of EMT, which may be related to the regulation of cellular calcium ions.