Department of Oncology, Beichen District Tradicine Chinese Medicine Hospital Affiliated to Tianjin University of Traditional Chinese Medicine, 300400 Tianjin, China.
Department of Orthopedics, Beichen District Tradicine Chinese Medicine Hospital Affiliated to Tianjin University of Traditional Chinese Medicine, 300400 Tianjin, China.
Discov Med. 2024 Aug;36(187):1648-1656. doi: 10.24976/Discov.Med.202436187.151.
Osteosarcoma (OS) is commonly recognized as a malignant cancer originating from bone-forming mesenchymal stem cells, comprising approximately 20% of sarcomas. Baicalin, a bioactive flavonoid glycoside isolated from , has been demonstrated to possess potent anti-inflammatory and neuroprotective properties.
To explore the potential mechanisms through which baicalin exerts anti-osteosarcoma effects and facilitates osteogenesis .
Cell Counting Kit-8 (CCK-8), scratch assay, and transwell assay were employed to assess the effects of baicalin at varying concentrations (20, 40, and 80 μM) on U2OS cell proliferation, invasion, and migration, respectively. Western blot and qRT-PCR analyses were conducted to evaluate the influence of baicalin on the osteogenic potential of OS cells by examining osteoblast markers such as osteocalcin (), osteopontin (), and runt-related transcription factor 2 (), as well as the osteoclast marker-receptor activator of nuclear factor kappa B ligand (). Additionally, the impact of baicalin on epithelial-mesenchymal transition (EMT) markers (N-cadherin, E-cadherin, Vimentin) and proteins related to the Nuclear factor κB (NF-κB) signaling pathway (p-p65, p-IκBα, p65, IκBα) in OS cells was evaluated via western blot analysis. The activity and mineralization capacity of Alkaline Phosphatase (ALP) in baicalin-treated cells were examined through ALP staining and Alizarin Red S (ARS) staining.
Baicalin exhibited significant suppression of OS cell U2OS invasion ( < 0.01), migration ( < 0.01), and proliferation ( < 0.05) at various concentrations. Additionally, baicalin treatment notably increased the E-cadherin protein level, while decreasing the expression levels of Vimentin and N-cadherin proteins ( < 0.01), thus promoting EMT. Following baicalin treatment, there was a marked elevation in the protein and mRNA expression levels of RUNX2, OPN, and OCN, while the expression level of RANKL protein was reduced ( < 0.05), indicating enhanced osteogenic differentiation. The groups treated with baicalin exhibited higher ALP activity and mineralization ability ( < 0.01). Moreover, baicalin treatment significantly reduced the expression levels of p-IκBα and p-p65 proteins, as well as the ratios of p-IκBα/IκBα and p-p65/p65 ( < 0.01). These effects of baicalin were concentration-dependent, with higher concentrations yielding stronger effects.
, baicalin demonstrates anti-OS effects and facilitates osteogenic differentiation, potentially by inhibiting NF-κB pathway activity.
骨肉瘤（OS）通常被认为是一种起源于成骨间充质干细胞的恶性癌症，约占肉瘤的 20%。黄芩苷是从 中分离得到的一种生物活性黄酮类糖苷，具有很强的抗炎和神经保护作用。
探讨黄芩苷发挥抗骨肉瘤作用和促进成骨作用的潜在机制。
采用细胞计数试剂盒-8（CCK-8）、划痕实验和 Transwell 实验分别评估黄芩苷在不同浓度（20、40 和 80 μM）下对 U2OS 细胞增殖、侵袭和迁移的影响。通过 Western blot 和 qRT-PCR 分析评估黄芩苷对 OS 细胞成骨潜能的影响，检测成骨细胞标志物骨钙素（）、骨桥蛋白（）和 runt 相关转录因子 2（）以及破骨细胞标志物核因子κB 配体（）。此外，通过 Western blot 分析评估黄芩苷对 OS 细胞上皮-间充质转化（EMT）标志物（N-钙粘蛋白、E-钙粘蛋白、波形蛋白）和核因子κB（NF-κB）信号通路相关蛋白（p-p65、p-IκBα、p65、IκBα）的影响。通过 ALP 染色和茜素红 S（ARS）染色检测黄芩苷处理细胞中碱性磷酸酶（ALP）的活性和矿化能力。
黄芩苷在不同浓度下显著抑制 OS 细胞 U2OS 的侵袭（ < 0.01）、迁移（ < 0.01）和增殖（ < 0.05）。此外，黄芩苷处理后 E-钙粘蛋白蛋白水平明显升高，而波形蛋白和 N-钙粘蛋白蛋白表达水平降低（ < 0.01），促进 EMT。黄芩苷处理后，RUNX2、OPN 和 OCN 的蛋白和 mRNA 表达水平明显升高，而 RANKL 蛋白表达水平降低（ < 0.05），表明成骨分化增强。黄芩苷处理组的 ALP 活性和矿化能力较高（ < 0.01）。此外，黄芩苷处理后 p-IκBα 和 p-p65 蛋白表达水平以及 p-IκBα/IκBα 和 p-p65/p65 比值均显著降低（ < 0.01）。黄芩苷的这些作用呈浓度依赖性，浓度越高作用越强。
黄芩苷具有抗 OS 作用和促进成骨分化的作用，可能通过抑制 NF-κB 通路活性来实现。
