Department of General Biology, Center of Biological Sciences, State University of Londrina (UEL), Londrina 86057-970, Brazil.
Department of General Biology, Center of Biological Sciences, State University of Londrina (UEL), Londrina 86057-970, Brazil.
Toxicol In Vitro. 2022 Sep;83:105416. doi: 10.1016/j.tiv.2022.105416. Epub 2022 Jun 13.
The present study investigates the mechanisms underlying the in vitro antitumoral activity of cirsimarin (CIR 10 to 320 μM), a flavone extracted from the aerial parts of Scoparia dulcis L., on MCF-7 cells cultured in 2D and multicellular tumor spheroids (3D). CIR (from 40 μM) decreased cell viability in the resazurin assay and colony formation in the 2D model. In the same way, in the 3D model, CIR (from 40 μM) induced cell death (triple staining assay) and decreased spheroid integrity after 16 days with no induction of intracellular reactive species (CM-HDCFDA). In 2D, CIR decreased the invasion (transwell) and horizontal migration (wound healing), while in 3D, CIR diminished cell migration (ECM® gel) and induced DNA damage (comet assay) possibly related to cell death. CIR mediated antitumoral effects in 3D spheroids by negative modulation of genes associated with cell proliferation (CCND1, CCNA2, CDK2, CDK4, and TNF) and death (BCL-XL, BAX, CASP9, and BIRC5). BIRC5 and CDKs inhibitors have been proposed as versatile anticancer drugs, which makes our results quite interesting. TNF negative modulation may also be related to the downregulation of MMP9 and MMP11 and anti-migration/invasion of MCF-7 cells cultured in 2D and 3D models. These are relevant properties for long-term strategies to avoid metastasis and improve the prognosis of breast cancer.
本研究探讨了从 Scoparia dulcis L. 的地上部分提取的类黄酮西瑞马林（CIR 10 至 320 μM）在体外对 MCF-7 细胞的抗肿瘤活性的机制，这些细胞在 2D 和多细胞肿瘤球体（3D）中培养。CIR（从 40 μM）降低了 2D 模型中的细胞活力和 RESAZURIN 测定中的集落形成。同样，在 3D 模型中，CIR（从 40 μM）诱导细胞死亡（三重染色测定）并降低球体完整性，而不诱导细胞内活性物质（CM-HDCFDA）。在 2D 中，CIR 降低了侵袭（Transwell）和水平迁移（伤口愈合），而在 3D 中，CIR 减少了细胞迁移（ECM®凝胶）并诱导 DNA 损伤（彗星测定），这可能与细胞死亡有关。CIR 通过负向调节与细胞增殖（CCND1、CCNA2、CDK2、CDK4 和 TNF）和死亡（BCL-XL、BAX、CASP9 和 BIRC5）相关的基因，介导 3D 球体中的抗肿瘤作用。BIRC5 和 CDK 抑制剂已被提议作为多功能抗癌药物，这使得我们的结果非常有趣。TNF 的负向调节也可能与 MMP9 和 MMP11 的下调以及 2D 和 3D 模型中 MCF-7 细胞的迁移/侵袭减少有关。这些是避免转移和改善乳腺癌预后的长期策略的相关特性。
