水杨梅素对人骨肉瘤细胞系的抗癌活性涉及 G2/M 细胞周期阻滞、线粒体凋亡以及抑制细胞迁移和侵袭。

Anticancer activity of Fisetin against the human osteosarcoma cell lines involves G2/M cell cycle arrest, mitochondrial apoptosis and inhibition of cell migration and invasion.

机构信息

Department of Orthopaedics, the First Affiliated Hospital, Zhejiang University, Hangzhou, Zhejiang, China, 310003.

出版信息

J BUON. 2020 Mar-Apr;25(2):1022-1027.

PMID:32521901

Abstract

PURPOSE

Osteosarcoma is rare but fatal type of human malignancy. The high metastasis rate, late diagnosis, emergence of drug resistance against drugs such as doxorubicin, and the lack of therapeutic targets obstructs the treatment of osteosarcoma. The present investigation explores the anticancer properties of Fisetin against human osteosarcoma cells.

METHODS

The cell viability was determined by WST-1 assay. DAPI and Annexin V/propidium iodide (PI) assays were used for detection of apoptosis. Flow cytometry was used for the determination of osteosarcoma MG-63 cell distribution. Wound healing and transwell assays were used for cell migration and invasion. Western blotting was used for protein expression analysis.

RESULTS

The results showed that Fisetin inhibits the growth of the MG-63 cells in a dose-dependent manner. Fisetin showed an IC50 of 18 µM against the MG-63 cells. The growth inhibitory effects of Fisetin were mainly due to induction of apoptosis which was accompanied by enhancement of the capsase-3 and Bax and depletion of Bcl-2 expression. Fisetin treatment increased reactive oxygen species (ROS) from 100 in untreated to 220% at 36 µM and decreased mitochondrial membrane potential (MMP) levels from 100 in untreated to 21% at 36 µM. Fisetin also induced G2/M cell cycle arrest of the MG-63 cells and suppressed the expression of cyclin-B1. The wound healing and the transwell assay showed that Fisetin suppressed the migration and invasion of the MG-63 cells.

CONCLUSION

Taken together, Fisetin may find use as lead molecule in the osteosarcoma therapeutic development programmes.

摘要

目的

骨肉瘤是一种罕见但致命的人类恶性肿瘤。其高转移率、晚期诊断、对多柔比星等药物的耐药性以及缺乏治疗靶点，阻碍了骨肉瘤的治疗。本研究探讨了非瑟酮对人骨肉瘤细胞的抗癌特性。

方法

通过 WST-1 测定法测定细胞活力。使用 DAPI 和 Annexin V/碘化丙啶（PI）测定法检测细胞凋亡。流式细胞术用于测定骨肉瘤 MG-63 细胞的分布。划痕愈合和 Transwell 测定法用于检测细胞迁移和侵袭。Western blot 用于蛋白表达分析。

结果

结果表明，非瑟酮以剂量依赖性方式抑制 MG-63 细胞的生长。非瑟酮对 MG-63 细胞的 IC50 为 18 μM。非瑟酮的生长抑制作用主要归因于诱导细胞凋亡，伴随 caspase-3 和 Bax 的增强以及 Bcl-2 表达的耗竭。非瑟酮处理将未处理组中的活性氧（ROS）从 100 增加到 36 μM 时的 220%，并将线粒体膜电位（MMP）水平从 100 降低到 36 μM 时的 21%。非瑟酮还诱导 MG-63 细胞的 G2/M 细胞周期停滞，并抑制细胞周期蛋白 B1 的表达。划痕愈合和 Transwell 测定法表明，非瑟酮抑制了 MG-63 细胞的迁移和侵袭。