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Abstract：
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.
摘要：
本研究旨在探讨雌激素受体β(ERβ)对骨肉瘤细胞的影响,并探索参与这一过程的调控机制。骨肉瘤U2-OS细胞分为四组，并通过E2，E2LY294002（ERβ激动剂）治疗，E2+ERβsiRNA,分别为E2+ERβsiRNA+LY294002。进行细胞计数试剂盒8(CCK-8)测定以检测各组U2-OS细胞的细胞活力。通过伤口愈合实验和transwell细胞培养室检测ERβ对U2-OS细胞迁移和侵袭能力的影响，分别。采用定量RT-PCR检测各组U2-OS细胞凋亡抑制蛋白(IAP)和整合素α5的表达,和磷酸化p65（p-p65）的表达，通过蛋白质印迹法检测p-AKT和Bcl-2。细胞活力，ERβsiRNA可显著提高U2-OS细胞的迁移和侵袭能力,但被ERβ激动剂LY294002抑制(p<0.05)。ERβsiRNA在U2-OS细胞中显著下调整合素α5和未调节的IAP(p<0.05)。LY294002可显著降低p-p65、p-AKT和Bcl-2的表达,而ERβsiRNA可显著升高(p<0.05)。总之,ERβ通过调节整合素对骨肉瘤细胞有明显的抗肿瘤作用,IAP,NF-kBBCL-2和PI3K/Akt旌旗灯号通路。
