Dependence of reactive oxygen species and FLICE inhibitory protein on Lipofectamine-induced apoptosis in human lung epithelial cells

Cationic liposomes such as Lipofectamine (LF) are widely used as nonviral gene delivery vectors; however, their clinical application is limited by their cytotoxicity. These agents have been shown to induce apoptosis as the primary mode of cell death, but their mechanism of action is not well underst...

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Main Authors: Lalana Kongkaneramit, Narong Sarisuta, Neelam Azad, Yongju Lu, Anand Krishnan V. Iyer, Liying Wang, Yon Rojanasakul
Other Authors: West Virginia University
Format: Article
Published: 2018
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Online Access:https://repository.li.mahidol.ac.th/handle/123456789/18913
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spelling th-mahidol.189132018-07-12T09:50:49Z Dependence of reactive oxygen species and FLICE inhibitory protein on Lipofectamine-induced apoptosis in human lung epithelial cells Lalana Kongkaneramit Narong Sarisuta Neelam Azad Yongju Lu Anand Krishnan V. Iyer Liying Wang Yon Rojanasakul West Virginia University Mahidol University National Institute for Occupational Safety and Health West Virginia University Robert C. Byrd Health Sciences Center Biochemistry, Genetics and Molecular Biology Pharmacology, Toxicology and Pharmaceutics Cationic liposomes such as Lipofectamine (LF) are widely used as nonviral gene delivery vectors; however, their clinical application is limited by their cytotoxicity. These agents have been shown to induce apoptosis as the primary mode of cell death, but their mechanism of action is not well understood. The present study investigated the mechanism of LF-induced apoptosis and examined the role of reactive oxygen species (ROS) in this process. We found that LF induced apoptosis of human epithelial H460 cells through a mechanism that involves caspase activation and ROS generation. Inhibition of caspase activity by pan-caspase inhibitor (z-VAD-fmk) or by specific caspase-8 inhibitor (z-IETD-fmk) or caspase-9 inhibitor (z-LEHD-fmk) inhibited the apoptotic effect of LF. Overexpression of FLICE-inhibitory protein (FLIP) or B-cell lymphoma-2, which are known inhibitors of the extrinsic and intrinsic death pathways, respectively, similarly inhibited apoptosis by LF. Induction of apoptosis by LF was shown to require ROS generation because its inhibition by ROS scavengers or by ectopic expression of antioxidant enzyme superoxide dismutase and glutathione peroxidase strongly inhibited the apoptotic effect of LF. Electron spin resonance studies showed that LF induced multiple ROS; however, superoxide was found to be the primary ROS responsible for LF-induced apoptosis. The mechanism by which ROS mediate the apoptotic effect of LF involves down-regulation of FLIP through the ubiquitination pathway. In demonstrating the role of FLIP and ROS in LF death signaling, we document a novel mechanism of apoptosis regulation that may be exploited to decrease cytotoxicity and increase gene transfection efficiency of cationic liposomes. 2018-07-12T02:18:28Z 2018-07-12T02:18:28Z 2008-06-01 Article Journal of Pharmacology and Experimental Therapeutics. Vol.325, No.3 (2008), 969-977 10.1124/jpet.107.136077 15210103 00223565 2-s2.0-44249116184 https://repository.li.mahidol.ac.th/handle/123456789/18913 Mahidol University SCOPUS https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=44249116184&origin=inward
institution Mahidol University
building Mahidol University Library
continent Asia
country Thailand
Thailand
content_provider Mahidol University Library
collection Mahidol University Institutional Repository
topic Biochemistry, Genetics and Molecular Biology
Pharmacology, Toxicology and Pharmaceutics
spellingShingle Biochemistry, Genetics and Molecular Biology
Pharmacology, Toxicology and Pharmaceutics
Lalana Kongkaneramit
Narong Sarisuta
Neelam Azad
Yongju Lu
Anand Krishnan V. Iyer
Liying Wang
Yon Rojanasakul
Dependence of reactive oxygen species and FLICE inhibitory protein on Lipofectamine-induced apoptosis in human lung epithelial cells
description Cationic liposomes such as Lipofectamine (LF) are widely used as nonviral gene delivery vectors; however, their clinical application is limited by their cytotoxicity. These agents have been shown to induce apoptosis as the primary mode of cell death, but their mechanism of action is not well understood. The present study investigated the mechanism of LF-induced apoptosis and examined the role of reactive oxygen species (ROS) in this process. We found that LF induced apoptosis of human epithelial H460 cells through a mechanism that involves caspase activation and ROS generation. Inhibition of caspase activity by pan-caspase inhibitor (z-VAD-fmk) or by specific caspase-8 inhibitor (z-IETD-fmk) or caspase-9 inhibitor (z-LEHD-fmk) inhibited the apoptotic effect of LF. Overexpression of FLICE-inhibitory protein (FLIP) or B-cell lymphoma-2, which are known inhibitors of the extrinsic and intrinsic death pathways, respectively, similarly inhibited apoptosis by LF. Induction of apoptosis by LF was shown to require ROS generation because its inhibition by ROS scavengers or by ectopic expression of antioxidant enzyme superoxide dismutase and glutathione peroxidase strongly inhibited the apoptotic effect of LF. Electron spin resonance studies showed that LF induced multiple ROS; however, superoxide was found to be the primary ROS responsible for LF-induced apoptosis. The mechanism by which ROS mediate the apoptotic effect of LF involves down-regulation of FLIP through the ubiquitination pathway. In demonstrating the role of FLIP and ROS in LF death signaling, we document a novel mechanism of apoptosis regulation that may be exploited to decrease cytotoxicity and increase gene transfection efficiency of cationic liposomes.
author2 West Virginia University
author_facet West Virginia University
Lalana Kongkaneramit
Narong Sarisuta
Neelam Azad
Yongju Lu
Anand Krishnan V. Iyer
Liying Wang
Yon Rojanasakul
format Article
author Lalana Kongkaneramit
Narong Sarisuta
Neelam Azad
Yongju Lu
Anand Krishnan V. Iyer
Liying Wang
Yon Rojanasakul
author_sort Lalana Kongkaneramit
title Dependence of reactive oxygen species and FLICE inhibitory protein on Lipofectamine-induced apoptosis in human lung epithelial cells
title_short Dependence of reactive oxygen species and FLICE inhibitory protein on Lipofectamine-induced apoptosis in human lung epithelial cells
title_full Dependence of reactive oxygen species and FLICE inhibitory protein on Lipofectamine-induced apoptosis in human lung epithelial cells
title_fullStr Dependence of reactive oxygen species and FLICE inhibitory protein on Lipofectamine-induced apoptosis in human lung epithelial cells
title_full_unstemmed Dependence of reactive oxygen species and FLICE inhibitory protein on Lipofectamine-induced apoptosis in human lung epithelial cells
title_sort dependence of reactive oxygen species and flice inhibitory protein on lipofectamine-induced apoptosis in human lung epithelial cells
publishDate 2018
url https://repository.li.mahidol.ac.th/handle/123456789/18913
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