Rosmarinic acid exerts an anticancer effect on osteosarcoma cells by inhibiting DJ-1 via regulation of the PTEN-PI3K-Akt signaling pathway
Rosmarinic acid exerts an anticancer effect on osteosarcoma cells by inhibiting DJ-1 via regulation of the PTEN-PI3K-Akt signaling pathway
复制标题
迷迭香酸通过调节PTEN-PI 3 K-Akt信号通路抑制DJ-1对骨肉瘤细胞的抗肿瘤作用
DOI：
10.1016/j.phymed.2020.153186
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发表时间：
2020-03-01
期刊：
影响因子：
7.9
通讯作者：
Lu, Li
中科院分区：
文献类型：
--
作者：
Ma, Zhanjun;Yang, Jingjing;Lu, Li
背景：骨肉瘤是最常见的原发恶性骨肿瘤。在过去的几十年里，这种疾病的存活率越来越低，这导致它成为人类死亡的主要原因。迷迭香酸(RA)是一种水溶性多酚类植物化学物质，对多种癌症有很强的抗癌作用，但其对骨肉瘤的潜在作用尚不清楚。方法：采用CCK-8实验、流式细胞仪分析、创伤愈合实验、Transwell实验、蛋白质组学分析以及shRNAs的应用，分析RA对U2OS和MG63骨肉瘤细胞存活、凋亡、细胞周期分布、迁移、侵袭和信号分子的影响。结果：RA对U2OS和MG63骨肉瘤细胞具有抗增殖和促凋亡作用。细胞凋亡通过增加Bax/Bcl2比值、触发细胞内活性氧产生、降低线粒体膜电位、上调caspase-8、caspase-9和caspase-3的裂解率来诱导细胞凋亡。此外，RA通过抑制基质金属蛋白酶-2和-9(MMP-2和-9)的表达水平来抑制骨肉瘤细胞的迁移和侵袭，这两种表达水平与上皮-间充质转化(EMT)的减弱有关。此外，蛋白质组学分析确定DJ-1是RA的潜在靶点。一些研究表明，通过慢病毒介导的shRNA转基因，DJ-1具有致癌作用，这导致明显抑制细胞的增殖、迁移和侵袭，并阻止细胞周期进展。在分子水平上，DJ-1、p-PI3K和p-Akt的表达水平降低，而磷酸酶和紧张素同源蛋白(PTEN)的蛋白水平升高。结论：结合DJ-1在骨肉瘤组织和细胞系中的高水平表达，本研究结果提示RA通过调节PTEN-PI3K-Akt信号通路抑制DJ-1，从而发挥抗骨肉瘤作用。因此，DJ-1可能是RA在骨肉瘤细胞中的生物学靶点。
Background: Osteosarcoma is the most common type of primary malignant bone tumor. This disease has exhibited a progressively lower survival rate over the past several decades, which has resulted in it becoming a main cause of death in humans. Rosmarinic acid (RA), a water-soluble polyphenolic phytochemical, exerts powerful anticancer effects against multiple types of cancer; however, its potential effects on osteosarcoma remain unknown. Hence, the present study investigated the efficacy of RA against osteosarcoma and aimed to clarify the mechanisms underlying this process.Methods: The effects of RA on cell viability, apoptosis, cell cycle distribution, migration, invasion, and signaling molecules were analyzed by CCK-8 assay, flowcytometric analysis, wound healing assay, Transwell assay, proteomic analysis, and use of shRNAs.Results: RA exerted anti-proliferation and pro-apoptotic effects on U2OS and MG63 osteosarcoma cells. Apoptosis was induced via extrinsic and intrinsic pathways by increasing the Bax/Bcl-2 ratio, triggering the intracellular production of reactive oxygen species (ROS), reducing the mitochondrial membrane potential (MMP), and upregulating the cleavage rates of caspase-8, caspase-9, and caspase-3. Additionally, RA suppressed the migration and invasion of osteosarcoma cells by inhibiting the expression levels of matrix metalloproteinase-2 and -9 (MMP-2 and -9), which are associated with a weakening of the epithelial-mesenchymal transition (EMT). Moreover, proteomic analyses identified DJ-1 as a potential target for RA. Several studies have indicated an oncogenic role for DJ-1 using knockdowns via the lentiviral-mediated transfection of shRNA, which caused the conspicuous suppression of cell proliferation, migration, and invasion as well as the arrest of cell cycle progression. At the molecular level, the expression levels of DJ-1, p-PI3K, and p-Akt were reduced, whereas the protein levels of phosphatase and tensin homologue (PTEN) were increased.Conclusion: In conjunction with the high levels of DJ-1 expression in osteosarcoma tissues and cell lines, the present results suggested that RA exhibited anticancer effects in osteosarcoma cells by inhibiting DJ-1 via regulation of the PTEN-PI3K-Akt signaling pathway. Therefore, DJ-1 might be a biological target for RA in osteosarcoma cells.