Investigation of the Apoptotic Effect of Levofloxacin on Ovarian Follicles of NMRI Mice and its Anticancer Effect on Human Ovarian Cancer Cell Line

Abstract

Background: Levofloxacin is a fluoroquinolone antibiotic with extensive anti-bacterial effects. Levofloxacin is widely used for treatment of urinary and vaginal infections. Objectives: This research surveyed the cytotoxicity effects of levofloxacin on ovarian follicles of mice in both in vitro and in vivo conditions, as well as its anticancer effect on human ovarian cancer cell line (SKOV3). Methods: For in vitro study, the ovaries of animals were isolated and treated with levofloxacin at doses of 1, 2, 5, and 10 µg/ml for 6 days. For in vivo study, animals were treated with levofloxacin at concentrations of 100, 200, 400, and 800 mg/kg for 24 days. Histopathological and morphological examinations of ovarian tissues were performed. Real-time PCR and Western Blot techniques were performed to analyze apoptosis-related genes and proteins of ovarian tissues. Results: Levofloxacin at higher concentrations caused morphological changes and remarkably decreased the number of primary, secondary, and adult follicles compared to the control group. The percentage of viable SKOV3 cells was 10.12%, 7.63%, and 2.17% following exposure to levofloxacin (at a concentration of 800 μg/ml) for 24, 48, and 72 h, respectively. The half-maximal inhibitory concentration of levofloxacin against SKOV3 was found to be 181.1, 74.84, and 27.58 μg/ml at 24, 48, and 72 h, respectively. The percentage of SKOV3 cells apoptosis following exposure to levofloxacin after 72 h at 20, 80, and 200 μg/ml doses was 11%, 42%, and 52%, respectively. Real-time PCR revealed up-regulation of Bax and Caspase-3 genes after exposure to levofloxacin in SKOV3 cells, whereas the expression of Bcl2 was significantly decreased in a concentration-depended manner. Conclusion: The present study demonstrates that levofloxacin induces apoptosis of both ovarian follicles and human ovarian cancer cell lines.