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Inhibition of Inducible Nitric Oxide Synthase (iNOS) by Andrographolide and In Vitro Evaluation of Its Antiproliferative and Proapoptotic Effects on Cervical Cancer

This work is aimed at investigating the expression levels of inducible nitric oxide synthase (iNOS) in cervical cancer and identifying a potential iNOS inhibitor. The data mining studies performed advocated iNOS to be a promising biomarker for cancer prognosis, as it is highly overexpressed in sever...

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Autores principales: Pasha, Akbar, Kumbhakar, Divya Vishambhar, Doneti, Ravinder, Kumar, Kiran, Dharmapuri, Gangappa, Poleboyina, Pavan Kumar, S. K., Heena, Basavaraju, Preethi, Pasumarthi, Deepthi, S. D., Annapurna, Soujanya, Pavani, Arnold Emeson, I., Bodiga, Vijayalaxmi, Pawar, Smita C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8010528/
https://www.ncbi.nlm.nih.gov/pubmed/33815659
http://dx.doi.org/10.1155/2021/6692628
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author Pasha, Akbar
Kumbhakar, Divya Vishambhar
Doneti, Ravinder
Kumar, Kiran
Dharmapuri, Gangappa
Poleboyina, Pavan Kumar
S. K., Heena
Basavaraju, Preethi
Pasumarthi, Deepthi
S. D., Annapurna
Soujanya, Pavani
Arnold Emeson, I.
Bodiga, Vijayalaxmi
Pawar, Smita C.
author_facet Pasha, Akbar
Kumbhakar, Divya Vishambhar
Doneti, Ravinder
Kumar, Kiran
Dharmapuri, Gangappa
Poleboyina, Pavan Kumar
S. K., Heena
Basavaraju, Preethi
Pasumarthi, Deepthi
S. D., Annapurna
Soujanya, Pavani
Arnold Emeson, I.
Bodiga, Vijayalaxmi
Pawar, Smita C.
author_sort Pasha, Akbar
collection PubMed
description This work is aimed at investigating the expression levels of inducible nitric oxide synthase (iNOS) in cervical cancer and identifying a potential iNOS inhibitor. The data mining studies performed advocated iNOS to be a promising biomarker for cancer prognosis, as it is highly overexpressed in several malignant cancers. The elevated iNOS was found to be associated with poor survival and increased tumor aggressiveness in cervical cancer. Immunohistochemical and RT-PCR investigations of iNOS showed significant upregulation of endogenous iNOS expression in the cervical tumor samples, thus making iNOS a potent target for decreasing tumor inflammation and aggressiveness. Andrographolide, a plant-derived diterpenoid lactone, is widely reported to be effective against infections and inflammation, causing no adverse side effects on humans. In the current study, we investigated the effect of andrographolide on the prognostic value of iNOS expression in cervical cancer, which has not been reported previously. The binding efficacy of andrographolide was analyzed by performing molecular docking and molecular dynamic simulations. Multiple parameters were used to analyze the simulation trajectory, like root mean square deviation (RMSD), torsional degree of freedom, protein-root mean square fluctuations (P-RMSF), ligand RMSF, total number of intramolecular hydrogen bonds, secondary structure elements (SSE) of the protein, and protein complex with the time-dependent functions of MDS. Ligand-protein interactions revealed binding efficacy of andrographolide with tryptophan amino acid of iNOS protein. Cancer cell proliferation, cell migration, cell cycle analysis, and apoptosis-mediated cell death were assessed in vitro, post iNOS inhibition induced by andrographolide treatment (demonstrated by Western blot). Results. Andrographolide exhibited cytotoxicity by inhibiting the in vitro proliferation of cervical cancer cells and also abrogated the cancer cell migration. A significant increase in apoptosis was observed with increasing andrographolide concentration, and it also induced cell cycle arrest at G1-S phase transition. Our results substantiate that andrographolide significantly inhibits iNOS expression and exhibits antiproliferative and proapoptotic effects on cervical cancer cells.
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spelling pubmed-80105282021-04-02 Inhibition of Inducible Nitric Oxide Synthase (iNOS) by Andrographolide and In Vitro Evaluation of Its Antiproliferative and Proapoptotic Effects on Cervical Cancer Pasha, Akbar Kumbhakar, Divya Vishambhar Doneti, Ravinder Kumar, Kiran Dharmapuri, Gangappa Poleboyina, Pavan Kumar S. K., Heena Basavaraju, Preethi Pasumarthi, Deepthi S. D., Annapurna Soujanya, Pavani Arnold Emeson, I. Bodiga, Vijayalaxmi Pawar, Smita C. Oxid Med Cell Longev Research Article This work is aimed at investigating the expression levels of inducible nitric oxide synthase (iNOS) in cervical cancer and identifying a potential iNOS inhibitor. The data mining studies performed advocated iNOS to be a promising biomarker for cancer prognosis, as it is highly overexpressed in several malignant cancers. The elevated iNOS was found to be associated with poor survival and increased tumor aggressiveness in cervical cancer. Immunohistochemical and RT-PCR investigations of iNOS showed significant upregulation of endogenous iNOS expression in the cervical tumor samples, thus making iNOS a potent target for decreasing tumor inflammation and aggressiveness. Andrographolide, a plant-derived diterpenoid lactone, is widely reported to be effective against infections and inflammation, causing no adverse side effects on humans. In the current study, we investigated the effect of andrographolide on the prognostic value of iNOS expression in cervical cancer, which has not been reported previously. The binding efficacy of andrographolide was analyzed by performing molecular docking and molecular dynamic simulations. Multiple parameters were used to analyze the simulation trajectory, like root mean square deviation (RMSD), torsional degree of freedom, protein-root mean square fluctuations (P-RMSF), ligand RMSF, total number of intramolecular hydrogen bonds, secondary structure elements (SSE) of the protein, and protein complex with the time-dependent functions of MDS. Ligand-protein interactions revealed binding efficacy of andrographolide with tryptophan amino acid of iNOS protein. Cancer cell proliferation, cell migration, cell cycle analysis, and apoptosis-mediated cell death were assessed in vitro, post iNOS inhibition induced by andrographolide treatment (demonstrated by Western blot). Results. Andrographolide exhibited cytotoxicity by inhibiting the in vitro proliferation of cervical cancer cells and also abrogated the cancer cell migration. A significant increase in apoptosis was observed with increasing andrographolide concentration, and it also induced cell cycle arrest at G1-S phase transition. Our results substantiate that andrographolide significantly inhibits iNOS expression and exhibits antiproliferative and proapoptotic effects on cervical cancer cells. Hindawi 2021-03-16 /pmc/articles/PMC8010528/ /pubmed/33815659 http://dx.doi.org/10.1155/2021/6692628 Text en Copyright © 2021 Akbar Pasha et al. https://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Pasha, Akbar
Kumbhakar, Divya Vishambhar
Doneti, Ravinder
Kumar, Kiran
Dharmapuri, Gangappa
Poleboyina, Pavan Kumar
S. K., Heena
Basavaraju, Preethi
Pasumarthi, Deepthi
S. D., Annapurna
Soujanya, Pavani
Arnold Emeson, I.
Bodiga, Vijayalaxmi
Pawar, Smita C.
Inhibition of Inducible Nitric Oxide Synthase (iNOS) by Andrographolide and In Vitro Evaluation of Its Antiproliferative and Proapoptotic Effects on Cervical Cancer
title Inhibition of Inducible Nitric Oxide Synthase (iNOS) by Andrographolide and In Vitro Evaluation of Its Antiproliferative and Proapoptotic Effects on Cervical Cancer
title_full Inhibition of Inducible Nitric Oxide Synthase (iNOS) by Andrographolide and In Vitro Evaluation of Its Antiproliferative and Proapoptotic Effects on Cervical Cancer
title_fullStr Inhibition of Inducible Nitric Oxide Synthase (iNOS) by Andrographolide and In Vitro Evaluation of Its Antiproliferative and Proapoptotic Effects on Cervical Cancer
title_full_unstemmed Inhibition of Inducible Nitric Oxide Synthase (iNOS) by Andrographolide and In Vitro Evaluation of Its Antiproliferative and Proapoptotic Effects on Cervical Cancer
title_short Inhibition of Inducible Nitric Oxide Synthase (iNOS) by Andrographolide and In Vitro Evaluation of Its Antiproliferative and Proapoptotic Effects on Cervical Cancer
title_sort inhibition of inducible nitric oxide synthase (inos) by andrographolide and in vitro evaluation of its antiproliferative and proapoptotic effects on cervical cancer
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8010528/
https://www.ncbi.nlm.nih.gov/pubmed/33815659
http://dx.doi.org/10.1155/2021/6692628
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