Department of Orthopedics, Traditional Chinese Medicine Hospital of Binzhou, Binzhou, Shandong 256600, P.R. China.
Department of Nursing, Traditional Chinese Medicine Hospital of Binzhou, Binzhou, Shandong 256600, P.R. China.
Mol Med Rep. 2022 Oct;26(4). doi: 10.3892/mmr.2022.12815. Epub 2022 Aug 5.
Hydroxygenkwanin (HGK) has an anticancer effect in a variety of tumors, but its role in osteosarcoma has not been explored. The purpose of the present study was to investigate the therapeutic effect of HGK on osteosarcoma and its specific molecular mechanism. Osteosarcoma cells (MG‑63 and U2OS) treated with various concentrations of HGK were assigned to the treatment group. MTT, clone formation, wound healing and Transwell assays were performed to assess the viability, proliferation, migration, and invasion of MG‑63 and U2OS cells. RT‑qPCR was conducted to quantify the expression levels of of microRNA (miR)‑320a and SRY‑box transcription factor 9 (SOX9) in MG‑63 and U2OS cells. The binding sites of miR‑320a and SOX9 were predicted by starBase database, and verified using the dual‑luciferase reporter assay. The expression levels of SOX9 and EMT‑related proteins (N‑cadherin, E‑cadherin and vimentin) were detected by western blot analysis. HGK inhibited cell proliferation, migration, invasion, but promoted the expression of miR‑320a in MG‑63 and U2OS cells. Downregulation of miR‑320a reversed the effects of HGK on proliferation, migration and invasion of MG‑63 and U2OS cells, while upregulation of miR‑320a had the opposite effect. HGK inhibited the expression of SOX9 by promoting the expression of miR‑320a. Upregulation of SOX9 could partially reverse miR‑320a‑induced migration and invasion of MG‑63 and U2OS cells. In addition, upregulation of miR‑320a promoted E‑cadherin expression and inhibited the expression of N‑cadherin and vimentin, and the effect of miR‑320a was also reversed by SOX9. In conclusion, HGK inhibited proliferation, migration and invasion of MG‑63 and U2OS cells through the miR‑320a/SOX9 axis.
Hydroxygenkwanin (HGK) 在多种肿瘤中具有抗癌作用，但它在骨肉瘤中的作用尚未得到探索。本研究旨在探讨 HGK 对骨肉瘤的治疗作用及其特定的分子机制。用不同浓度的 HGK 处理骨肉瘤细胞（MG-63 和 U2OS），将其分为治疗组。通过 MTT、克隆形成、划痕愈合和 Transwell 实验评估 MG-63 和 U2OS 细胞的活力、增殖、迁移和侵袭。通过 RT-qPCR 定量测定 MG-63 和 U2OS 细胞中 microRNA（miR）-320a 和性别决定区 Y 框转录因子 9（SOX9）的表达水平。miR-320a 和 SOX9 的结合位点通过 starBase 数据库预测，并通过双荧光素酶报告基因实验验证。Western blot 分析检测 SOX9 和 EMT 相关蛋白（N-钙黏蛋白、E-钙黏蛋白和波形蛋白）的表达水平。HGK 抑制细胞增殖、迁移和侵袭，但促进 MG-63 和 U2OS 细胞中 miR-320a 的表达。下调 miR-320a 逆转了 HGK 对 MG-63 和 U2OS 细胞增殖、迁移和侵袭的影响，而上调 miR-320a 则产生相反的效果。HGK 通过促进 miR-320a 的表达来抑制 SOX9 的表达。上调 SOX9 可部分逆转 miR-320a 诱导的 MG-63 和 U2OS 细胞的迁移和侵袭。此外，上调 miR-320a 促进 E-钙黏蛋白的表达，抑制 N-钙黏蛋白和波形蛋白的表达，而 SOX9 也逆转了 miR-320a 的作用。综上所述，HGK 通过 miR-320a/SOX9 轴抑制 MG-63 和 U2OS 细胞的增殖、迁移和侵袭。
