Harbin Medical University, Harbin, Heilongjiang, China (mainland).
Department of Oncological Surgery, The First Hospital of Qiqihaer City, Qiqihaer, Heilongjiang, China (mainland).
Med Sci Monit. 2018 Jul 16;24:4926-4933. doi: 10.12659/MSM.909193.
BACKGROUND The aim of this study was investigate the effects of the sesquiterpene lactone, ludartin, on cell proliferation, cell migration, apoptosis, and the cell cycle in osteosarcoma cell lines, compared with a normal osteoblast cell line. MATERIAL AND METHODS Osteosarcoma cell lines, MG-63 Saos-2 U-2OS, T1-73 143B, and HOS, and normal hFOB 1.19 osteoblasts, were cultured and treated with increasing doses of ludartin, The MTT colorimetric assay was used to measure cell metabolic activity and viability. Apoptosis was studied by fluorescence-activated cell sorting (FACS) using 4',6-diamidino-2-phenylindole (DAPI) nuclear staining and Annexin-V/propidium iodide (PI) staining. Cell cycle was studied using flow cytometry. Cell migration and invasion were studied using wound healing and Boyden chamber assays. Protein expression was measured by Western blotting. RESULTS Ludartin inhibited cell viability, cell migration, cell proliferation, and increased cell apoptosis, in all osteosarcoma cell lines, with an IC50 dose ranging from 15-30 µM. The greatest effects were on the Saso-2 osteosarcoma cells, with an IC50 of 15 µM. However, ludartin showed minor cytotoxic effects of the normal hFOB 1.19 osteoblasts (IC50 >100 µM). Ludartin exerted its anti-proliferative effects on Saos-2 cells via induction of apoptosis and cell cycle arrest at the G2/M checkpoint, associated with reduced expression of Cdc25c (Ser216), Cdc25c, pCdc2 (Tyr15), and Cdc2 and increased expression of p21WAF1. Ludartin inhibited cell migration and invasion of the Saos-2 cells. CONCLUSIONS The dose-dependent effects of ludartin on cell proliferation, migration, apoptosis, cell cycle arrest at the G2/M checkpoint involved p21WAFI in Saos-2 osteosarcoma cells.
本研究旨在探讨倍半萜内酯 ludartin 对骨肉瘤细胞系与正常成骨细胞系细胞增殖、细胞迁移、凋亡和细胞周期的影响。
培养骨肉瘤细胞系 MG-63、Saos-2、U-2OS、T1-73、143B 和 HOS，以及正常 hFOB 1.19 成骨细胞，并使用递增剂量的 ludartin 处理细胞。使用 MTT 比色法测定细胞代谢活性和活力。通过使用 4'，6-二脒基-2-苯基吲哚（DAPI）核染色和 Annexin-V/碘化丙啶（PI）染色，通过荧光激活细胞分选（FACS）研究凋亡。使用流式细胞术研究细胞周期。通过划痕愈合和 Boyden 室测定研究细胞迁移和侵袭。通过 Western 印迹测定测量蛋白表达。
Ludartin 抑制了所有骨肉瘤细胞系的细胞活力、细胞迁移、细胞增殖，并增加了细胞凋亡，IC50 剂量范围为 15-30 µM。在 Saso-2 骨肉瘤细胞中作用最大，IC50 为 15 µM。然而，ludartin 对正常 hFOB 1.19 成骨细胞的细胞毒性较小（IC50 > 100 µM）。Ludartin 通过诱导细胞凋亡和细胞周期阻滞在 G2/M 检查点，使 Saos-2 细胞中的增殖受到抑制，与 Cdc25c（Ser216）、Cdc25c、pCdc2（Tyr15）和 Cdc2 的表达减少以及 p21WAF1 的表达增加有关。Ludartin 抑制了 Saos-2 细胞的迁移和侵袭。
Ludartin 对细胞增殖、迁移、凋亡和 G2/M 检查点细胞周期阻滞的剂量依赖性影响涉及 Saos-2 骨肉瘤细胞中的 p21WAF1。
