Lan Haifeng, Hong Wei, Fan Pan, Qian Dongyang, Zhu Jianwei, Bai Bo
Department of Orthopaedic Surgery, 1st Affiliated Hospital of Guangzhou Medical University, Guangdong Provincial Key Lab of Orthopaedic Technology and Implant Materials, Guangzhou, China.
GMU-GIBH Joint School of Life Sciences, Guangzhou Medical University, Guangzhou, China.
Cell Physiol Biochem. 2017;43(2):553-567. doi: 10.1159/000480528. Epub 2017 Sep 21.
BACKGROUND/AIMS: Osteosarcoma is a malignant tumor associated with high mortality; however, no effective therapies for the disease have been developed. Several studies have focused on elucidating the pathogenesis of osteosarcoma and have aimed to develop novel therapies for the disease. Quercetin is a vital dietary flavonoid that has been shown to have a variety of anticancer effects, as it induces cell cycle arrest, apoptosis, and differentiation and is involved in cell adhesion, metastasis and angiogenesis. Herein, we aimed to investigate the effects of quercetin on osteosarcoma migration and invasion in vitro and in vivo and to explore the molecular mechanisms underlying its effects on osteosarcoma migration and invasion.
Cell viability, cell cycle activity and cell apoptosis were measured using CCK-8 assay and flow cytometry, and cell migration and invasion were evaluated by wound healing and transwell assays, respectively. The mRNA and protein expression levels of several proteins of interest were assessed by real-time quantitative PCR and western blotting, respectively. Moreover, a nude mouse model of human osteosarcoma lung metastasis was established to assess the anti-metastatic effects of quercetin in vivo.
We noted no significant differences in cell cycle activity and apoptosis between HOS and MG63 cells and control cells. Treatment with quercetin significantly attenuated cell migration and invasion in HOS and MG63 cells compared with treatment with control medium. Moreover HIF-1α, VEGF, MMP2, and MMP9 mRNA and protein expression levels were significantly downregulated in HOS cells treated with quercetin compared with HOS cells treated with controls. Additionally, treatment with quercetin attenuated metastatic lung tumor formation and growth in the nude mouse model of osteosarcoma compared with treatment with controls.
Our findings regarding the inhibitory effects of quercetin on cell migration and invasion suggest that quercetin may have potential as a therapy for human osteosarcoma.
背景/目的:骨肉瘤是一种死亡率很高的恶性肿瘤;然而,尚未开发出针对该疾病的有效疗法。多项研究致力于阐明骨肉瘤的发病机制,并旨在开发针对该疾病的新疗法。槲皮素是一种重要的膳食类黄酮,已被证明具有多种抗癌作用,因为它可诱导细胞周期停滞、凋亡和分化,并参与细胞黏附、转移和血管生成。在此,我们旨在研究槲皮素在体外和体内对骨肉瘤迁移和侵袭的影响,并探讨其对骨肉瘤迁移和侵袭作用的分子机制。
使用CCK-8法和流式细胞术测量细胞活力、细胞周期活性和细胞凋亡,分别通过伤口愈合试验和Transwell试验评估细胞迁移和侵袭。分别通过实时定量PCR和蛋白质印迹法评估几种感兴趣蛋白质的mRNA和蛋白质表达水平。此外,建立人骨肉瘤肺转移裸鼠模型以评估槲皮素在体内的抗转移作用。
我们发现HOS和MG63细胞与对照细胞之间在细胞周期活性和凋亡方面无显著差异。与用对照培养基处理相比,用槲皮素处理显著减弱了HOS和MG63细胞的迁移和侵袭。此外,与用对照处理的HOS细胞相比,用槲皮素处理的HOS细胞中HIF-1α、VEGF、MMP2和MMP9的mRNA和蛋白质表达水平显著下调。另外,与用对照处理相比,用槲皮素处理减弱了骨肉瘤裸鼠模型中转移性肺肿瘤的形成和生长。
我们关于槲皮素对细胞迁移和侵袭抑制作用的研究结果表明,槲皮素可能具有作为人类骨肉瘤治疗方法的潜力。