Department of Biochemistry and Molecular Biology, Basic Medical College of Jiujiang University, No. 320, Xunyang East Road, Xunyang District, Jiujiang City, Jiangxi Province 332000, China.
Department of Biochemistry and Molecular Biology, Basic Medical College of Jiujiang University, No. 320, Xunyang East Road, Xunyang District, Jiujiang City, Jiangxi Province 332000, China.
Pathol Res Pract. 2019 Oct;215(10):152568. doi: 10.1016/j.prp.2019.152568. Epub 2019 Jul 27.
The present study aimed to explore the potential anti-tumor effect of ERβ overexpression and investigate its related mechanism in osteosarcoma. Cell cycle and apoptosis rates were measured by flow cytometry. Cell proliferation and formation of autophagosome were assessed by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay and dansylcadaverine (MDC) staining assay. Cell migration and invasion were detected by wound healing assay and transwell assay. Western blot analysis was designed to detect the protein expressions of surviving, Bax, LC-3 П, Beclin-1, ERβ, TβRⅠ, TβRⅡ, Smad2, Smad3 and Smad7. Real-Time fluorogenic PCR was designed to examine the mRNA expressions of surviving, Bax, ERβ, TβRⅠ, TβRII, Smad2, Smad3 and Smad7. The results showed that ERβ overexpression inhibited cell proliferation, migration and invasion, blocked cell cycle, and induced apoptosis and autophagy. Additionally, ERβ overexpression significantly inhibited the expression of surviving, TβRⅠ, TβRⅡ, Smad2 and Smad3. Meanwhile, the expressions of Bax, LC-3 П, Beclin-1 and Smad7 were dramatically upregulated by ERβ overexpression. In conclusion, ERβ overexpression could inhibit cell proliferation, migration and invasion, block cell cycle, and promote apoptosis and autophagy in OS by downregulating TNG-β signaling pathway.
本研究旨在探讨 ERβ 过表达在骨肉瘤中的潜在抗肿瘤作用，并研究其相关机制。采用流式细胞术检测细胞周期和细胞凋亡率。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)法和丹磺酰尸胺(MDC)染色法评估细胞增殖和自噬小体形成。通过划痕愈合试验和 Transwell 试验检测细胞迁移和侵袭。Western blot 分析用于检测存活蛋白、Bax、LC-3 П、Beclin-1、ERβ、TβRⅠ、TβRⅡ、Smad2、Smad3 和 Smad7 的蛋白表达。实时荧光定量 PCR 用于检测存活蛋白、Bax、ERβ、TβRⅠ、TβRⅡ、Smad2、Smad3 和 Smad7 的 mRNA 表达。结果表明，ERβ 过表达抑制细胞增殖、迁移和侵袭，阻断细胞周期，诱导细胞凋亡和自噬。此外，ERβ 过表达显著抑制存活蛋白、TβRⅠ、TβRⅡ、Smad2 和 Smad3 的表达。同时，ERβ 过表达显著上调 Bax、LC-3 П、Beclin-1 和 Smad7 的表达。总之，ERβ 过表达通过下调 TNG-β 信号通路，抑制 OS 细胞增殖、迁移和侵袭，阻断细胞周期，促进细胞凋亡和自噬。
