Department of Hematology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430030, China.
J BUON. 2020 May-Jun;25(3):1600-1606.
The purpose of the study was to examine the anticancer properties of a natural chemical entity - juglone molecule - against human HL-60 promyelocytic leukemia cells along with assessing the effects on normal human monocytes. Juglone molecule was examined for its role in autophagy induction, endogenous ROS production, and inhibition of cell migration and invasion.
Cell viability was evaluated by MTT assay and clonogenic assay was performed to analyse colony formation. Autophagy studies were carried out by transmission electron microscopy (TEM) and western blotting analysis. Mitochondrial morphology was observed through MitoTracker Red CMXRos, and mitochondrial ROS production was assessed through confocal microscopy. The cell invasion and migration was assessed via transwell chambers with and without Matrigel.
MTT assay revealed significant, selective (less cytotoxicity towards normal cells) and dose-dependent inhibition of HL-60 leukemia cells and clonogenic assay showed impressive decrease in the number of colonies on increased doses of the molecule. TEM analysis showed formation of autophagosomes and induction of cellular damage. Western blotting assay indicated a significant increase in LC3-I and LC3-II and a slight increase in Beclin-I. Confocal microscopy revealed tremendous increase in ROS concentrations in a dose-dependent manner. Transwell chamber assay revealed significant, dose- dependent inhibition of cell migration and invasion.
Juglone induced anticancer effects on promyelocytic HL-60 leukemia cells mediated via autophagy induction, endogenous ROS production, and inhibition of cell migration and invasion, thus indicating that juglone may be a potential lead molecule against HL-60 promyelocytic leukemia cells.
本研究旨在探讨天然化学物质 - 胡桃醌分子对人 HL-60 早幼粒细胞白血病细胞的抗癌特性，并评估其对正常人类单核细胞的影响。研究了胡桃醌分子在自噬诱导、内源性 ROS 产生以及抑制细胞迁移和侵袭中的作用。
通过 MTT 测定法评估细胞活力，并通过集落形成分析进行克隆形成测定。通过透射电子显微镜 (TEM) 和 Western 印迹分析进行自噬研究。通过 MitoTracker Red CMXRos 观察线粒体形态，通过共聚焦显微镜评估线粒体 ROS 产生。通过带有和不带有 Matrigel 的 Transwell 室评估细胞侵袭和迁移。
MTT 测定法显示 HL-60 白血病细胞的显著、选择性（对正常细胞的细胞毒性较小）和剂量依赖性抑制，克隆形成测定法显示随着分子剂量的增加，菌落数量明显减少。TEM 分析显示自噬体的形成和细胞损伤的诱导。Western 印迹分析表明 LC3-I 和 LC3-II 显著增加，Beclin-I 略有增加。共聚焦显微镜显示 ROS 浓度呈剂量依赖性显著增加。Transwell 室测定法显示细胞迁移和侵袭的显著、剂量依赖性抑制。
胡桃醌通过诱导自噬、产生内源性 ROS 以及抑制细胞迁移和侵袭，对早幼粒细胞 HL-60 白血病细胞产生抗癌作用，表明胡桃醌可能是一种针对 HL-60 早幼粒细胞白血病细胞的潜在先导分子。
