Department of Oncology, The First Affiliated Hospital of Gannan Medical University, Ganzhou, Jiangxi, China.
Department of Orthopedics, The First Affiliated Hospital of Gannan Medical University, Ganzhou, Jiangxi, China.
J Orthop Surg Res. 2020 Sep 3;15(1):374. doi: 10.1186/s13018-020-01879-0.
Osteosarcoma (OS) is a common severe illness globally. Lupeol has been reported to participate in the pathophysiologic properties of various cancers, including OS. This study aimed to explore the effects of lupeol on proliferation, invasion, and apoptosis on OS cells and the underlying mechanism.
The cell viability of OS cells was determined by 3-(4, 5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. The expression levels of miR-212-3p and high-mobility group AT-hook 2 (HMGA2) were detected by quantitative real-time polymerase chain reaction (qRT-PCR) in OS cells. The cell apoptosis and invasion were detected by flow cytometry and transwell invasion assays, respectively. The functional target of miR-212-3p was predicted by online software and confirmed by luciferase reporter assay. The protein level of HMGA2 was measured by western blot analysis.
Lupeol suppressed cell viability and invasion, and promoted apoptosis by upregulating the expression of miR-212-3p in OS cells. Knockdown of miR-212-3p restored the anti-tumor effect of lupeol. Interestingly, miR-212-3p directly targeted HMGA2 and suppressed its expression. Moreover, HMGA2 reversed the inhibited impact on viability and invasion, and the promoted effect on apoptosis induced by upregulation of miR-212-3p. Also, lupeol administration exerts its anti-tumor effect by overexpression of miR-212-3p to suppress the expression of HMGA2 in OS cells.
Lupeol inhibited OS progression by modulating the miR-212-3p/HMGA2 axis in vitro.
骨肉瘤(OS)是一种在全球范围内较为常见的严重疾病。已有报道称,羽扇醇参与了包括 OS 在内的多种癌症的病理生理特性。本研究旨在探讨羽扇醇对 OS 细胞增殖、侵袭和凋亡的影响及其潜在机制。
采用 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴盐(MTT)法测定 OS 细胞的细胞活力。采用实时定量聚合酶链反应(qRT-PCR)检测 OS 细胞中 miR-212-3p 和高迁移率族蛋白 A2(HMGA2)的表达水平。采用流式细胞术和 Transwell 侵袭实验分别检测细胞凋亡和侵袭。通过在线软件预测 miR-212-3p 的功能靶标,并通过荧光素酶报告基因实验进行验证。采用 Western blot 分析检测 HMGA2 蛋白水平。
羽扇醇通过上调 OS 细胞中 miR-212-3p 的表达抑制细胞活力和侵袭,促进细胞凋亡。miR-212-3p 的敲低恢复了羽扇醇的抗肿瘤作用。有趣的是,miR-212-3p 直接靶向 HMGA2 并抑制其表达。此外,HMGA2 逆转了 miR-212-3p 上调诱导的对活力和侵袭的抑制作用,以及对凋亡的促进作用。此外,羽扇醇通过上调 miR-212-3p 抑制 HMGA2 的表达,从而发挥其在 OS 细胞中的抗肿瘤作用。
羽扇醇通过调节 miR-212-3p/HMGA2 轴在体外抑制 OS 进展。