Cargando…
Autophagic Vacuolation Induced by Excess ROS Generation in HABP1/p32/gC1qR Overexpressing Fibroblasts and Its Reversal by Polymeric Hyaluronan
The ubiquitous hyaladherin, hyaluronan-binding protein 1 (HABP1/p32/gC1qR) upon stable overexpression in normal fibroblasts (F-HABP07) has been reported to induce mitochondrial dysfunction, growth retardation and apoptosis after 72 h of growth. HABP1 has been observed to accumulate in the mitochondr...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2013
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3799741/ https://www.ncbi.nlm.nih.gov/pubmed/24205125 http://dx.doi.org/10.1371/journal.pone.0078131 |
_version_ | 1782287918663991296 |
---|---|
author | Saha, Paramita Chowdhury, Anindya Roy Dutta, Shubhra Chatterjee, Soumya Ghosh, Ilora Datta, Kasturi |
author_facet | Saha, Paramita Chowdhury, Anindya Roy Dutta, Shubhra Chatterjee, Soumya Ghosh, Ilora Datta, Kasturi |
author_sort | Saha, Paramita |
collection | PubMed |
description | The ubiquitous hyaladherin, hyaluronan-binding protein 1 (HABP1/p32/gC1qR) upon stable overexpression in normal fibroblasts (F-HABP07) has been reported to induce mitochondrial dysfunction, growth retardation and apoptosis after 72 h of growth. HABP1 has been observed to accumulate in the mitochondria resulting in generation of excess Reactive Oxygen Species (ROS), mitochondrial Ca(++) efflux and drop in mitochondrial membrane potential. In the present study, autophagic vacuolation was detected with monodansylcadaverin (MDC) staining from 36 h to 60 h of culture period along with elevated level of ROS in F-HABP07 cells. Increased expression of autophagic markers like MAP-LC3-II, Beclin 1 and autophagic modulator, DRAM confirmed the occurrence of the phenomenon. Reduced vacuole formation was observed upon treatment with 3-MA, a known PI3 kinase inhibitor, only at 32 h and was ineffective if treated later, as high ROS level was already attained. Treatment of F111 and F-HABP07 cells with bafilomycin A1 further indicated an increase in autophagosome formation along with autophagic degradation in HABP1 overexpressed fibroblasts. Comparison between normal fibroblast (F111) and F-HABP07 cells indicate reduced level of polymeric HA, its depolymerization and perturbed HA-HABP1 interaction in F-HABP07. Interestingly, supplementation of polymeric HA, an endogenous ROS scavenger, in the culture medium prompted reduction in number of vacuoles in F-HABP07 along with drop in ROS level, implying that excess ROS generation triggers initiation of autophagic vacuole formation prior to apoptosis due to overexpression of HABP1. Thus, the phenomenon of autophagy takes place prior to apoptosis induction in the HABP1 overexpressing cell line, F-HABP07. |
format | Online Article Text |
id | pubmed-3799741 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-37997412013-11-07 Autophagic Vacuolation Induced by Excess ROS Generation in HABP1/p32/gC1qR Overexpressing Fibroblasts and Its Reversal by Polymeric Hyaluronan Saha, Paramita Chowdhury, Anindya Roy Dutta, Shubhra Chatterjee, Soumya Ghosh, Ilora Datta, Kasturi PLoS One Research Article The ubiquitous hyaladherin, hyaluronan-binding protein 1 (HABP1/p32/gC1qR) upon stable overexpression in normal fibroblasts (F-HABP07) has been reported to induce mitochondrial dysfunction, growth retardation and apoptosis after 72 h of growth. HABP1 has been observed to accumulate in the mitochondria resulting in generation of excess Reactive Oxygen Species (ROS), mitochondrial Ca(++) efflux and drop in mitochondrial membrane potential. In the present study, autophagic vacuolation was detected with monodansylcadaverin (MDC) staining from 36 h to 60 h of culture period along with elevated level of ROS in F-HABP07 cells. Increased expression of autophagic markers like MAP-LC3-II, Beclin 1 and autophagic modulator, DRAM confirmed the occurrence of the phenomenon. Reduced vacuole formation was observed upon treatment with 3-MA, a known PI3 kinase inhibitor, only at 32 h and was ineffective if treated later, as high ROS level was already attained. Treatment of F111 and F-HABP07 cells with bafilomycin A1 further indicated an increase in autophagosome formation along with autophagic degradation in HABP1 overexpressed fibroblasts. Comparison between normal fibroblast (F111) and F-HABP07 cells indicate reduced level of polymeric HA, its depolymerization and perturbed HA-HABP1 interaction in F-HABP07. Interestingly, supplementation of polymeric HA, an endogenous ROS scavenger, in the culture medium prompted reduction in number of vacuoles in F-HABP07 along with drop in ROS level, implying that excess ROS generation triggers initiation of autophagic vacuole formation prior to apoptosis due to overexpression of HABP1. Thus, the phenomenon of autophagy takes place prior to apoptosis induction in the HABP1 overexpressing cell line, F-HABP07. Public Library of Science 2013-10-18 /pmc/articles/PMC3799741/ /pubmed/24205125 http://dx.doi.org/10.1371/journal.pone.0078131 Text en © 2013 Saha et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Saha, Paramita Chowdhury, Anindya Roy Dutta, Shubhra Chatterjee, Soumya Ghosh, Ilora Datta, Kasturi Autophagic Vacuolation Induced by Excess ROS Generation in HABP1/p32/gC1qR Overexpressing Fibroblasts and Its Reversal by Polymeric Hyaluronan |
title | Autophagic Vacuolation Induced by Excess ROS Generation in HABP1/p32/gC1qR Overexpressing Fibroblasts and Its Reversal by Polymeric Hyaluronan |
title_full | Autophagic Vacuolation Induced by Excess ROS Generation in HABP1/p32/gC1qR Overexpressing Fibroblasts and Its Reversal by Polymeric Hyaluronan |
title_fullStr | Autophagic Vacuolation Induced by Excess ROS Generation in HABP1/p32/gC1qR Overexpressing Fibroblasts and Its Reversal by Polymeric Hyaluronan |
title_full_unstemmed | Autophagic Vacuolation Induced by Excess ROS Generation in HABP1/p32/gC1qR Overexpressing Fibroblasts and Its Reversal by Polymeric Hyaluronan |
title_short | Autophagic Vacuolation Induced by Excess ROS Generation in HABP1/p32/gC1qR Overexpressing Fibroblasts and Its Reversal by Polymeric Hyaluronan |
title_sort | autophagic vacuolation induced by excess ros generation in habp1/p32/gc1qr overexpressing fibroblasts and its reversal by polymeric hyaluronan |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3799741/ https://www.ncbi.nlm.nih.gov/pubmed/24205125 http://dx.doi.org/10.1371/journal.pone.0078131 |
work_keys_str_mv | AT sahaparamita autophagicvacuolationinducedbyexcessrosgenerationinhabp1p32gc1qroverexpressingfibroblastsanditsreversalbypolymerichyaluronan AT chowdhuryanindyaroy autophagicvacuolationinducedbyexcessrosgenerationinhabp1p32gc1qroverexpressingfibroblastsanditsreversalbypolymerichyaluronan AT duttashubhra autophagicvacuolationinducedbyexcessrosgenerationinhabp1p32gc1qroverexpressingfibroblastsanditsreversalbypolymerichyaluronan AT chatterjeesoumya autophagicvacuolationinducedbyexcessrosgenerationinhabp1p32gc1qroverexpressingfibroblastsanditsreversalbypolymerichyaluronan AT ghoshilora autophagicvacuolationinducedbyexcessrosgenerationinhabp1p32gc1qroverexpressingfibroblastsanditsreversalbypolymerichyaluronan AT dattakasturi autophagicvacuolationinducedbyexcessrosgenerationinhabp1p32gc1qroverexpressingfibroblastsanditsreversalbypolymerichyaluronan |