Departments of Traditional Chinese Medical Orthopedics, Affiliated Hospital of Shandong Academy of Medical Sciences, Ji'nan, Shandong 250031, P.R. China.
Department of Orthopedics, Qilu Hospital of Shandong University, Ji'nan, Shandong 250012, P.R. China.
Biosci Rep. 2018 Dec 14;38(6). doi: 10.1042/BSR20181501. Print 2018 Dec 21.
Osteosarcoma (OS) is the most common primary malignancy of skeleton with higher mortality rates amongst children and young adults worldwide, whereas effective and secure therapies have also been sought by researches with ongoing efforts. The purpose of the present study was to investigate the impact of N'-[(3Z)-1-(1-hexyl)-2-oxo-1,2-dihydro-3H-indol-3-ylidene] benzohydrazide (MDA19) on OS and explore its potential mechanism. Cell Counting Kit-8 (CCK8) and colony formation assay were employed to evaluate the potential effect of MDA19 on U2OS and MG-63 cells proliferation. Moreover, transwell migration and invasion assay were performed to assess the influence of MDA19 on U2OS and MG-63 cells migration and invasion. In addition, Annexin V-FITC/propidium iodide (Annexin V-FITC/PI) staining and flow cytometry were used to examine apoptotic ratio of the U2OS and MG-63 cells. Meanwhile, Western blot analysis was applied to explore change of relevant mechanism proteins in OS cells treated with MDA19. Our study showed that MDA19 had anti-proliferative activity of OS cells in a dose- and time-dependent manner, simultaneously, inhibition of colony formation was also observed in U2OS and MG-63 cells after incubation of MDA19. Besides, MDA19 could significantly inhibit the number of migrated and invaded OS cells and markedly increase the OS cells apoptosis rate. Mechanistically, we detected detectable reductions in apoptosis related proteins, epithelial-mesenchymal transition (EMT)-related proteins and activity of phosphatidylinositol 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) signaling in U2OS and MG-63 cells exposure to MDA19. Overall, the current study indicates anti-proliferative, anti-metastatic, and pro-apoptotic potential of MDA19 in U2OS and MG-63 cells. Our findings propose a clue for further studies with this compound in preclinical and clinical treatment for OS.
骨肉瘤（OS）是骨骼中最常见的原发性恶性肿瘤，全球儿童和青年的死亡率较高，而研究人员也一直在寻求有效和安全的治疗方法。本研究旨在探讨 N'-[(3Z)-1-(1-己基)-2-氧代-1,2-二氢-3H-吲哚-3-亚基]苯甲酰肼（MDA19）对 OS 的影响，并探讨其潜在机制。细胞计数试剂盒-8（CCK8）和集落形成实验用于评估 MDA19 对 U2OS 和 MG-63 细胞增殖的潜在影响。此外，Transwell 迁移和侵袭实验用于评估 MDA19 对 U2OS 和 MG-63 细胞迁移和侵袭的影响。此外，使用 Annexin V-FITC/碘化丙啶（Annexin V-FITC/PI）染色和流式细胞术检测 U2OS 和 MG-63 细胞的凋亡率。同时，Western blot 分析用于研究 MDA19 处理后的 OS 细胞中相关机制蛋白的变化。我们的研究表明，MDA19 对 OS 细胞具有剂量和时间依赖性的抗增殖活性，同时，MDA19 孵育后 U2OS 和 MG-63 细胞中的集落形成也受到抑制。此外，MDA19 可显著抑制迁移和侵袭的 OS 细胞数量，并显著增加 OS 细胞的凋亡率。在机制上，我们检测到 MDA19 处理后 U2OS 和 MG-63 细胞中凋亡相关蛋白、上皮-间充质转化（EMT）相关蛋白和磷脂酰肌醇 3-激酶（PI3K）/Akt/雷帕霉素靶蛋白（mTOR）信号的活性明显降低。总的来说，本研究表明 MDA19 在 U2OS 和 MG-63 细胞中具有抗增殖、抗转移和促凋亡作用。我们的研究结果为进一步研究该化合物在骨肉瘤的临床前和临床治疗提供了线索。
