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

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

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.