Anticancer Effects of Juglone in OVCAR-3 Human Ovarian Carcinoma Are Facilitated Through Programmed Cell Death, Endogenous ROS Production, Inhibition of Cell Migration and Invasion and Cell Cycle Arrest

J BUON. Mar-Apr 2020;25(2):779-784.

Abstract

Purpose: Accumulating evidence suggests that Juglone is a potent anticancer molecule of plant origin. However, its anticancer effects have not been fully explored against human ovarian cancer cells. Therefore this study was undertaken to evaluate the anticancer effects of Juglone against the human OVCAR-3 ovarian cancer cells.

Methods: Cell viability was evaluated by WST-1 assay. Cellular apoptosis was studied using fluorescence microscopy with DAPI staining. The percentage of OVCAR-3 human ovarian cancer cells was examined by using flow cytometry in combination with annexin V-FITC/propidium iodide (PI) staining. Effects on cell cycle were studied by flow cytometer while effects on cell migration and invasion were evaluated using wound healing and transwell assay, respectively.

Results: Juglone inhibited the growth rate of OVCAR-3 ovarian cancer cells and showed an IC50 of 30 µM. However, Juglone showed very high IC50 (100 µM) against the normal SV40 ovarian cells. DAPI staining showed that Juglone caused nuclear fragmentation of the OVCAR-3 cells, suggestive of apoptosis. Annexin V/PI staining showed that the percentage of the apoptotic OVCAR-3 cells increased from 2.15 in control to 45.24% at 60 µM concentration of Juglone. The induction of apoptosis in the OVCAR-3 cells was also accompanied with activation caspase-3, upregulation of Bax and downregulation of Bcl-2. Juglone was also found to cause an upsurge in the ROS levels in the OVCAR-3 cells. Cell cycle analysis showed that Juglone caused accumulation of the OVCAR-3 cells in the G2/M phase of the cell cycle triggering G2/M cell cycle arrest. Wound healing assay and transwell assay showed that Juglone suppressed the migration as well as the invasion of the OVCAR-3 cells, suggestive of the antimetastatic potential of this molecule.

Conclusions: Juglone may prove advantageous in ovarian cancer treatment.