Low concentrations of doxycycline attenuates FasL-induced apoptosis in HeLa cells

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 μg/mL. However, the effect of DC in apoptotic signaling at much low concentrations was not stud...

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Autores principales:	Yoon,Jung Mi, Koppula,Sushruta, Huh,Se Jong, Hur,Sun Jin, Kim,Chan Gil
Lenguaje:	English
Publicado:	Sociedad de Biología de Chile 2015
Materias:	Apoptosis Caspase Cisplatin FasL Hydrogen peroxide Tetracycline
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602015000100038
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spelling	oai:scielo:S0716-976020150001000382016-02-16Low concentrations of doxycycline attenuates FasL-induced apoptosis in HeLa cellsYoon,Jung MiKoppula,SushrutaHuh,Se JongHur,Sun JinKim,Chan Gil Apoptosis Caspase Cisplatin FasL Hydrogen peroxide Tetracycline 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 μg/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 μg/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 μg/mL) significantly (p < 0.001) attenuated the FasL-induced cell death as measured by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazo-lium 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 μg/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 μg/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 cas-pase-8 and caspase-3, were inhibited by DC treatment at low concentration (0.5 μg/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.info:eu-repo/semantics/openAccessSociedad de Biología de ChileBiological Research v.48 20152015-01-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602015000100038en10.1186/S40659-015-0025-8
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	Apoptosis Caspase Cisplatin FasL Hydrogen peroxide Tetracycline
spellingShingle	Apoptosis Caspase Cisplatin FasL Hydrogen peroxide Tetracycline Yoon,Jung Mi Koppula,Sushruta Huh,Se Jong Hur,Sun Jin Kim,Chan Gil Low concentrations of doxycycline attenuates FasL-induced apoptosis in HeLa cells
description	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 μg/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 μg/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 μg/mL) significantly (p < 0.001) attenuated the FasL-induced cell death as measured by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazo-lium 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 μg/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 μg/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 cas-pase-8 and caspase-3, were inhibited by DC treatment at low concentration (0.5 μg/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.
author	Yoon,Jung Mi Koppula,Sushruta Huh,Se Jong Hur,Sun Jin Kim,Chan Gil
author_facet	Yoon,Jung Mi Koppula,Sushruta Huh,Se Jong Hur,Sun Jin Kim,Chan Gil
author_sort	Yoon,Jung Mi
title	Low concentrations of doxycycline attenuates FasL-induced apoptosis in HeLa cells
title_short	Low concentrations of doxycycline attenuates FasL-induced apoptosis in HeLa cells
title_full	Low concentrations of doxycycline attenuates FasL-induced apoptosis in HeLa cells
title_fullStr	Low concentrations of doxycycline attenuates FasL-induced apoptosis in HeLa cells
title_full_unstemmed	Low concentrations of doxycycline attenuates FasL-induced apoptosis in HeLa cells
title_sort	low concentrations of doxycycline attenuates fasl-induced apoptosis in hela cells
publisher	Sociedad de Biología de Chile
publishDate	2015
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602015000100038
work_keys_str_mv	AT yoonjungmi lowconcentrationsofdoxycyclineattenuatesfaslinducedapoptosisinhelacells AT koppulasushruta lowconcentrationsofdoxycyclineattenuatesfaslinducedapoptosisinhelacells AT huhsejong lowconcentrationsofdoxycyclineattenuatesfaslinducedapoptosisinhelacells AT hursunjin lowconcentrationsofdoxycyclineattenuatesfaslinducedapoptosisinhelacells AT kimchangil lowconcentrationsofdoxycyclineattenuatesfaslinducedapoptosisinhelacells
_version_	1718441536897679360

Low concentrations of doxycycline attenuates FasL-induced apoptosis in HeLa cells

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