Abstract：
Salvianolic acid B (Sal B) has demonstrated anticancer activity against various types of cancer. However, the underlying mechanism of Sal B-mediated anticancer effects remains incompletely understood. This study aims to investigate the impact of Sal B on the growth and metastasis of human A549 lung cells, as well as elucidate its potential mechanisms. In this study, different concentrations of Sal B were administered to A549 cells. The effects on migration and invasion abilities were assessed using MTT, wound healing, and transwell assays. Flow cytometry analysis was employed to evaluate Sal B-induced apoptosis in A549 cells. Western blotting and immunohistochemistry were conducted to measure the expression levels of cleaved caspase-3, cleaved PARP, and E-cadherin. Commercial kits were utilized for detecting intracellular reactive oxygen species (ROS) and NAD+. Additionally, a xenograft model with transplanted A549 tumors was employed to assess the anti-tumor effect of Sal B in vivo. The expression levels of NDRG2, p-PTEN, and p-AKT were determined through western blotting. Our findings demonstrate that Sal B effectively inhibits proliferation, migration, and invasion in A549 cells while inducing dose-dependent apoptosis. These apoptotic responses and inhibition of tumor cell metastasis are accompanied by alterations in intracellular ROS levels and NAD+/NADH ratio. Furthermore, our in vivo experiment reveals that Sal B significantly suppresses A549 tumor growth compared to an untreated control group while promoting increased cleavage of caspase-3 and PARP. Importantly, we observe that Sal B upregulates NDRG2 expression while downregulating p-PTEN and p-AKT expressions. Collectively, our results provide compelling evidence supporting the ability of Sal B to inhibit both growth and metastasis in A549 lung cancer cells through oxidative stress modulation as well as involvement of the NDRG2/PTEN/AKT pathway.
摘要：
丹酚酸B（SalB）已证明对各种类型的癌症具有抗癌活性。然而,SalB介导的抗癌作用的潜在机制仍未完全了解。本研究旨在探讨SalB对人A549肺细胞生长和转移的影响,以及阐明其潜在的机制。在这项研究中,对A549细胞施用不同浓度的SalB。使用MTT评估对迁移和入侵能力的影响，伤口愈合，和transwell分析。流式细胞术分析用于评估SalB诱导的A549细胞凋亡。进行蛋白质印迹和免疫组织化学以测量裂解的caspase-3，裂解的PARP的表达水平，和E-cadherin.商业试剂盒用于检测细胞内活性氧(ROS)和NAD+。此外,使用具有移植的A549肿瘤的异种移植模型来评估SalB在体内的抗肿瘤作用。NDRG2,p-PTEN的表达水平,通过蛋白质印迹法测定p-AKT。我们的研究结果表明，SalB有效抑制增殖，迁移,和侵袭A549细胞，同时诱导剂量依赖性凋亡。这些凋亡反应和肿瘤细胞转移的抑制伴随着细胞内ROS水平和NAD+/NADH比率的改变。此外，我们的体内实验显示,与未处理的对照组相比,SalB显著抑制A549肿瘤生长,同时促进caspase-3和PARP的裂解增加.重要的是,我们观察到SalB上调NDRG2表达，同时下调p-PTEN和p-AKT表达。总的来说，我们的结果提供了令人信服的证据,支持SalB通过氧化应激调节以及NDRG2/PTEN/AKT通路的参与抑制A549肺癌细胞的生长和转移.
