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Bcl-2相关抗凋亡蛋白3(Bcl-2 associated athanogene 3,BAG3)是BAG家族的重要成员,调节肿瘤细胞的粘附、迁移和侵袭,促进恶性肿瘤的复发和转移.本室前期工作证明,PKCδ可催化BAG3的Ser187位点磷酸化.本文研究BAG3蛋白磷酸化修饰对甲状腺癌FRO细胞EMT表型转化的影响.稳定转染野生型WT-BAG3、模拟磷酸化型S187D-BAG3、阻碍磷酸化型S187A-BAG3 FRO细胞后,观察细胞形态的变化.结果显示,稳定转染模拟磷酸化型S187D-BAG3引起甲状腺癌FRO细胞呈现明显的间质细胞形态.实时PCR和Western印迹,结果显示,稳定表达S187D-BAG3显著上调间质细胞标记物N-cadherin和波形蛋白mRNA与蛋白质在FRO细胞的表达,但下调上皮细胞标记物E-cadherin的mRNA和蛋白质的表达.同时,免疫荧光结果显示,稳定过表达S187D-BAG3的FRO细胞,E-cadherin和β-catenin出现向核周的内化.本文结果提示,BAG3蛋白磷酸化修饰可诱导甲状腺癌FRO细胞上皮间质转化.
Bcl-2 associated athanogene 3 (BAG3), an important member of BAG family, regulates the adhesion, migration and invasion of tumor cells and promotes the recurrence and metastasis of malignant tumors. , PKCδ can catalyze the phosphorylation of Ser187 at BAG3.This study was designed to investigate the effect of BAG3 phosphorylation on the phenotype of EMT phenotype in thyroid cancer cell line FRO.Transfect wild type WT-BAG3, mimic phosphorylated S187D-BAG3, S187A-BAG3 FRO cells were treated with S187A-BAG3 FRO cells.The results showed that stable transfection of S187D-BAG3 induced obvious morphological changes of stromal cells in thyroid cancer cells by real-time PCR and Western blotting.Results: Stable expression of S187D-BAG3 significantly upregulated the expression of interstitial cell markers N-cadherin and vimentin mRNA and protein in FRO cells, but downregulated the expression of E-cadherin mRNA and protein in epithelial cells.At the same time, immunofluorescence results showed that S187D- FRO cells stably overexpressing S187D-BAG3, E-cadherin and β-catenin appear to the perinuclear internalization.The results suggest that BAG3 protein phosphorylation can induce thyroid cancer FRO epithelial cells Mesenchymal transition.