Low concentrations of doxycycline attenuates FasL-induced apoptosis in HeLa cellsopen access
- Authors
- Yoon, Jung Mi; Koppula, Sushruta; Huh, Se Jong; Hur, Sun Jin; Kim, Chan Gil
- Issue Date
- Jul-2015
- Publisher
- SOC BIOLGIA CHILE
- Keywords
- Apoptosis; Caspase; Cisplatin; FasL; Hydrogen peroxide; Tetracycline
- Citation
- BIOLOGICAL RESEARCH, v.48
- Journal Title
- BIOLOGICAL RESEARCH
- Volume
- 48
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/9334
- DOI
- 10.1186/s40659-015-0025-8
- ISSN
- 0716-9760
0717-6287
- Abstract
- Background: Doxycycline (DC) has been shown to possess non-antibiotic properties including Fas/Fas Ligand (FasL)-mediated apoptosis against several tumor types in the concentration range of 10-40 mu g/mL. However, the effect of DC in apoptotic signaling at much low concentrations was not studied. Methods: The present study investigated the attenuation effect of low dose of DC on FasL-induced apoptosis in HeLa cell by the methods of MTT assay, fluorescence microscopy, DNA fragmentation, flow cytometry analysis, and western blotting. Results and conclusion: In the present findings we showed that low concentration of DC (<2.0 mu g/mL) exhibited protective effects against FasL-induced apoptosis in HeLa cells. FasL treatment to HeLa cells resulted in a concentration-dependent induction of cell death, and treatment with low concentrations of DC (0.1-2 mu g/mL) significantly (p < 0.001) attenuated the FasL-induced cell death as measured by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Further, the FasL-induced apoptotic features in HeLa cells, such as morphological changes, DNA fragmentation and cell cycle arrest was also inhibited by DC (0.5 mu g/mL). Tetracycline and minocycline also showed similar anti-apoptotic effects but were not significant when compared to DC, tested at same concentrations. Further, DC (0.01-16 mu g/mL) did not influence the hydrogen peroxide-or cisplatin-induced intrinsic apoptotic pathway in HeLa cells. Protein analysis using Western blotting confirmed that FasL-induced cleavage/activation of caspase-8 and caspase-3, were inhibited by DC treatment at low concentration (0.5 mu g/mL). Considering the overall data, we report for the first time that DC exhibited anti-apoptotic effects at low concentrations in HeLa cells by inhibition of caspase activation via FasL-induced extrinsic pathway.
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