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Interplay between Endoplasmic Reticular Stress and Survivin in Colonic Epithelial Cells

Sustained endoplasmic reticular stress (ERS) is implicated in aggressive metastasis of cancer cells and increased tumor cell proliferation. Cancer cells activate the unfolded protein response (UPR), which aids in cellular survival and adaptation to harsh conditions. Inhibition of apoptosis, in contr...

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Autores principales: Gundamaraju, Rohit, Vemuri, Ravichandra, Chong, Wai Chin, Myers, Stephen, Norouzi, Shaghayegh, Shastri, Madhur D., Eri, Rajaraman
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6210275/
https://www.ncbi.nlm.nih.gov/pubmed/30326660
http://dx.doi.org/10.3390/cells7100171
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author Gundamaraju, Rohit
Vemuri, Ravichandra
Chong, Wai Chin
Myers, Stephen
Norouzi, Shaghayegh
Shastri, Madhur D.
Eri, Rajaraman
author_facet Gundamaraju, Rohit
Vemuri, Ravichandra
Chong, Wai Chin
Myers, Stephen
Norouzi, Shaghayegh
Shastri, Madhur D.
Eri, Rajaraman
author_sort Gundamaraju, Rohit
collection PubMed
description Sustained endoplasmic reticular stress (ERS) is implicated in aggressive metastasis of cancer cells and increased tumor cell proliferation. Cancer cells activate the unfolded protein response (UPR), which aids in cellular survival and adaptation to harsh conditions. Inhibition of apoptosis, in contrast, is a mechanism adopted by cancer cells with the help of the inhibitor of an apoptosis (IAP) class of proteins such as Survivin to evade cell death and gain a proliferative advantage. In this study, we aimed to reveal the interrelation between ERS and Survivin. We initially verified the expression of Survivin in Winnie (a mouse model of chronic ERS) colon tissues by using immunohistochemistry (IHC) and immunofluorescence (IF) in comparison with wild type Blk6 mice. Additionally, we isolated the goblet cells and determined the expression of Survivin by IF and protein validation. Tunicamycin was utilized at a concentration of 10 µg/mL to induce ERS in the LS174T cell line and the gene expression of the ERS markers was measured. This was followed by determination of inflammatory cytokines. Inhibition of ERS was carried out by 4Phenyl Butyric acid (4PBA) at a concentration of 10 mM to assess whether there was a reciprocation effect. The downstream cell death assays including caspase 3/7, Annexin V, and poly(ADP-ribose) polymerase (PARP) cleavage were evaluated in the presence of ERS and absence of ERS, which was followed by a proliferative assay (EdU click) with and without ERS. Correspondingly, we inhibited Survivin by YM155 at a concentration of 100 nM and observed the succeeding ERS markers and inflammatory markers. We also verified the caspase 3/7 assay. Our results demonstrate that ERS inhibition not only significantly reduced the UPR genes (Grp78, ATF6, PERK and XBP1) along with Survivin but also downregulated the inflammatory markers such as IL8, IL4, and IL6, which suggests a positive correlation between ERS and the inhibition of apoptosis. Furthermore, we provided evidence that ERS inhibition promoted apoptosis in LS174T cells and shortened the proliferation rate. Moreover, Survivin inhibition by YM155 led to a comparable effect as that of ERS inhibition, which includes attenuation of ERS genes and inflammatory markers as well as the promotion of programmed cell death via the caspase 3/7 pathway. Together, our results propose the interrelation between ERS and inhibition of apoptosis assigning a molecular and therapeutic target for cancer treatment.
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spelling pubmed-62102752018-11-02 Interplay between Endoplasmic Reticular Stress and Survivin in Colonic Epithelial Cells Gundamaraju, Rohit Vemuri, Ravichandra Chong, Wai Chin Myers, Stephen Norouzi, Shaghayegh Shastri, Madhur D. Eri, Rajaraman Cells Article Sustained endoplasmic reticular stress (ERS) is implicated in aggressive metastasis of cancer cells and increased tumor cell proliferation. Cancer cells activate the unfolded protein response (UPR), which aids in cellular survival and adaptation to harsh conditions. Inhibition of apoptosis, in contrast, is a mechanism adopted by cancer cells with the help of the inhibitor of an apoptosis (IAP) class of proteins such as Survivin to evade cell death and gain a proliferative advantage. In this study, we aimed to reveal the interrelation between ERS and Survivin. We initially verified the expression of Survivin in Winnie (a mouse model of chronic ERS) colon tissues by using immunohistochemistry (IHC) and immunofluorescence (IF) in comparison with wild type Blk6 mice. Additionally, we isolated the goblet cells and determined the expression of Survivin by IF and protein validation. Tunicamycin was utilized at a concentration of 10 µg/mL to induce ERS in the LS174T cell line and the gene expression of the ERS markers was measured. This was followed by determination of inflammatory cytokines. Inhibition of ERS was carried out by 4Phenyl Butyric acid (4PBA) at a concentration of 10 mM to assess whether there was a reciprocation effect. The downstream cell death assays including caspase 3/7, Annexin V, and poly(ADP-ribose) polymerase (PARP) cleavage were evaluated in the presence of ERS and absence of ERS, which was followed by a proliferative assay (EdU click) with and without ERS. Correspondingly, we inhibited Survivin by YM155 at a concentration of 100 nM and observed the succeeding ERS markers and inflammatory markers. We also verified the caspase 3/7 assay. Our results demonstrate that ERS inhibition not only significantly reduced the UPR genes (Grp78, ATF6, PERK and XBP1) along with Survivin but also downregulated the inflammatory markers such as IL8, IL4, and IL6, which suggests a positive correlation between ERS and the inhibition of apoptosis. Furthermore, we provided evidence that ERS inhibition promoted apoptosis in LS174T cells and shortened the proliferation rate. Moreover, Survivin inhibition by YM155 led to a comparable effect as that of ERS inhibition, which includes attenuation of ERS genes and inflammatory markers as well as the promotion of programmed cell death via the caspase 3/7 pathway. Together, our results propose the interrelation between ERS and inhibition of apoptosis assigning a molecular and therapeutic target for cancer treatment. MDPI 2018-10-15 /pmc/articles/PMC6210275/ /pubmed/30326660 http://dx.doi.org/10.3390/cells7100171 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Gundamaraju, Rohit
Vemuri, Ravichandra
Chong, Wai Chin
Myers, Stephen
Norouzi, Shaghayegh
Shastri, Madhur D.
Eri, Rajaraman
Interplay between Endoplasmic Reticular Stress and Survivin in Colonic Epithelial Cells
title Interplay between Endoplasmic Reticular Stress and Survivin in Colonic Epithelial Cells
title_full Interplay between Endoplasmic Reticular Stress and Survivin in Colonic Epithelial Cells
title_fullStr Interplay between Endoplasmic Reticular Stress and Survivin in Colonic Epithelial Cells
title_full_unstemmed Interplay between Endoplasmic Reticular Stress and Survivin in Colonic Epithelial Cells
title_short Interplay between Endoplasmic Reticular Stress and Survivin in Colonic Epithelial Cells
title_sort interplay between endoplasmic reticular stress and survivin in colonic epithelial cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6210275/
https://www.ncbi.nlm.nih.gov/pubmed/30326660
http://dx.doi.org/10.3390/cells7100171
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