Yang Minfei, Liu Bing, Jin Libin, Tao Huimin, Yang Zhengming
Emergency Department, The Second Affiliated Hospital of Zhejiang University College of Medicine, Hangzhou, Zhejiang Province 310009, China.
Department of Orthopaedics, The Second Affiliated Hospital of Zhejiang University College of Medicine, Hangzhou, Zhejiang Province 310009, China.
J Bone Oncol. 2017 Sep 28;9:15-20. doi: 10.1016/j.jbo.2017.09.005. eCollection 2017 Nov.
This study aimed to investigate the effects of Estrogen receptor β (ERβ) on osteosarcoma cells, and explore the regulatory mechanisms involved in this process. Osteosarcoma U2-OS cells consisted four groups, and treated by E2, E2 + LY294002 (ERβ agonists), E2 + ERβ siRNA, E2 + ERβ siRNA + LY294002, respectively. Cell counting kit 8 (CCK-8) assay was performed to detect the cell viability of U2-OS cells in each group. The effects of ERβ on the migration and invasion ability of U2-OS cells were examined by wound healing assay and transwell cell culture chamber, respectively. The expression of Inhibitor of apoptosis protein (IAP) and integrin α5 in U2-OS cells of each group was detected by quantitative RT-PCR, and the expression of phosphorylated p65 (p-p65), p-AKT and Bcl-2 was detected by western blotting. The cell viability, migration and invasion ability of U2-OS cells were significantly increased by ERβ siRNA, but inhibited by ERβ agonists LY294002 (p < 0.05). ERβ siRNA significantly downregulated Integrin α5 and unregulated IAP in U2-OS cells (p < 0.05). The expression of p-p65, p-AKT and Bcl-2 was significantly reduced by LY294002, but increased by ERβ siRNA (p < 0.05). In conclusion, ERβ exhibited obvious anti-tumor effects on osteosarcoma cells by regulating integrin, IAP, NF-kBBCL-2 and PI3K/Akt signal pathway.
本研究旨在探讨雌激素受体β(ERβ)对骨肉瘤细胞的影响,并探索这一过程中涉及的调控机制。骨肉瘤U2-OS细胞分为四组,分别用E2、E2 + LY294002(ERβ激动剂)、E2 + ERβ siRNA、E2 + ERβ siRNA + LY294002处理。采用细胞计数试剂盒8(CCK-8)法检测各组U2-OS细胞的活力。分别通过伤口愈合试验和Transwell细胞培养小室检测ERβ对U2-OS细胞迁移和侵袭能力的影响。采用定量RT-PCR检测各组U2-OS细胞中凋亡抑制蛋白(IAP)和整合素α5的表达,采用蛋白质印迹法检测磷酸化p65(p-p65)、p-AKT和Bcl-2的表达。ERβ siRNA可显著提高U2-OS细胞的活力、迁移和侵袭能力,但ERβ激动剂LY294002可抑制这些能力(p < 0.05)。ERβ siRNA可显著下调U2-OS细胞中整合素α5的表达并上调IAP的表达(p < 0.05)。LY294002可显著降低p-p65、p-AKT和Bcl-2的表达,但ERβ siRNA可使其表达增加(p < 0.05)。综上所述,ERβ通过调节整合素、IAP、NF-kB/BCL-2和PI3K/Akt信号通路对骨肉瘤细胞表现出明显的抗肿瘤作用。