Young Researchers and Elite Club, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran.
Faculty of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran.
Anticancer Agents Med Chem. 2020;20(9):1087-1093. doi: 10.2174/1871520620666200408081111.
Osteosarcoma (OS) is known as the malignant tumors in the bone. Cyanidin 3-OGlucoside (C3G) has a potential to induce the apoptotic cell death in different cancer cells; however, the mechanisms of action for C3G have not been clarified yet.
In this study, the apoptotic effects of C3G on three different osteosarcoma cell lines including Saso-2, MG-63, and G-292 (clone A141B1) were investigated.
The 24-hr IC50 of C3G for Saso-2, G-292, and MG-63 cells was evaluated by the MTT assay. Apoptosis induction in these cell lines after treatment with the C3G was approved by the Annexin V/PI flow cytometry. Changes at the mRNA expression level of PPARγ, P21, Bax, and Bcl-xl genes were investigated by real-time Polymerase Chain Reaction (PCR) technique, and P21 expression was further confirmed by the western blotting.
The MTT assay results demonstrated that the 24-hr IC50 of C3G was equal to 110μg/ml for Saso-2 and G-292 cells while it was about 140μg/ml for the MG-63 cells. The results of real-time PCR clearly showed that treatment of the cells with 24hrs IC50 of C3G caused the upregulation of PPARγ, P21, and Bax genes. Moreover, western blot analysis confirmed that P21 protein overexpressed endogenously after treatment of the cells with the C3G, and it was more upregulated in the MG-63 cells compared to the other cell lines.
According to the findings of the study, the C3G is a novel anti-osteosarcoma agent with the ability to induce the apoptosis in different osteosarcoma cells through upregulation of the PPARγ and P21 genes.
骨肉瘤(OS)是一种已知的骨恶性肿瘤。矢车菊素 3-O-葡萄糖苷(C3G)具有诱导不同癌细胞凋亡的潜力;然而,C3G 的作用机制尚未阐明。
本研究旨在探讨 C3G 对三种不同骨肉瘤细胞系(包括 Saso-2、MG-63 和 G-292(克隆 A141B1))的凋亡作用。
通过 MTT 法评估 C3G 对 Saso-2、G-292 和 MG-63 细胞的 24 小时 IC50。通过 Annexin V/PI 流式细胞术证实 C3G 处理后这些细胞系的凋亡诱导。通过实时聚合酶链反应(PCR)技术研究 PPARγ、P21、Bax 和 Bcl-xl 基因的 mRNA 表达水平的变化,并通过蛋白质印迹法进一步证实 P21 的表达。
MTT 法结果表明,C3G 的 24 小时 IC50 对 Saso-2 和 G-292 细胞为 110μg/ml,而对 MG-63 细胞约为 140μg/ml。实时 PCR 结果清楚地表明,用 24 小时 IC50 的 C3G 处理细胞会导致 PPARγ、P21 和 Bax 基因的上调。此外,蛋白质印迹分析证实,用 C3G 处理细胞后,P21 蛋白内源性过表达,并且在 MG-63 细胞中比其他细胞系表达更高。
根据研究结果,C3G 是一种新型抗骨肉瘤药物,能够通过上调 PPARγ 和 P21 基因诱导不同骨肉瘤细胞凋亡。