Cargando…

Lipofection of Single Guide RNA Targeting MMP8 Decreases Proliferation and Migration in Lung Adenocarcinoma Cells

Background and Objectives: Matrix metalloproteinases (MMP) have been implicated as major determinants of tumour growth and metastasis, which are considered two of the main hallmarks of cancer. The interaction of MMP8 and other signalling molecules within and adjacent tumoral tissues, including immun...

Descripción completa

Detalles Bibliográficos
Autores principales: Maradiaga, Oscar David Hernandez, Mok, Pooi Ling, Sivapragasam, Gothai, Samrot, Antony V., Ali Khan, Mohammed Safwan, Farhana, Aisha, Alzahrani, Badr, Tong, Jiabei, Karuppiah, Thilakavathy, Joseph, Narcisse M. S., Subbiah, Suresh Kumar
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8306211/
https://www.ncbi.nlm.nih.gov/pubmed/34356991
http://dx.doi.org/10.3390/medicina57070710
_version_ 1783727755693654016
author Maradiaga, Oscar David Hernandez
Mok, Pooi Ling
Sivapragasam, Gothai
Samrot, Antony V.
Ali Khan, Mohammed Safwan
Farhana, Aisha
Alzahrani, Badr
Tong, Jiabei
Karuppiah, Thilakavathy
Joseph, Narcisse M. S.
Subbiah, Suresh Kumar
author_facet Maradiaga, Oscar David Hernandez
Mok, Pooi Ling
Sivapragasam, Gothai
Samrot, Antony V.
Ali Khan, Mohammed Safwan
Farhana, Aisha
Alzahrani, Badr
Tong, Jiabei
Karuppiah, Thilakavathy
Joseph, Narcisse M. S.
Subbiah, Suresh Kumar
author_sort Maradiaga, Oscar David Hernandez
collection PubMed
description Background and Objectives: Matrix metalloproteinases (MMP) have been implicated as major determinants of tumour growth and metastasis, which are considered two of the main hallmarks of cancer. The interaction of MMP8 and other signalling molecules within and adjacent tumoral tissues, including immune cells, are rather elusive, particularly of adenocarcinoma cell type. In this study, we aimed to investigate the role of MMP8 in non-small cell lung cancer proliferation and invasiveness potential. Materials and Methods: We individually lipofected with two different single guide RNA (sgRNAs) that specifically targeted on MMP8, with CRISPR-Cas 9 protein into the cells. Results: Our results clearly indicated that the lipofection of these complexes could lead to reduced ability of A549 cells to survive and proliferate to form colonies. In addition, when compared to non-transfected cells, the experimental cell groups receiving sgRNAs demonstrated relatively decreased migration rate, hence, wider wound gaps in scratch assay. The quantitative real time-polymerase chain reaction (qRT-PCR) demonstrated significant reduction in the MAP-K, survivin and PI3-K gene expression. MMP8 might have protective roles over tumour growth and spread in our body. Conclusions: The delivery of sgRNAs targeting on the MMP8 gene could induce tumour cell death and arrest cell migratory activity.
format Online
Article
Text
id pubmed-8306211
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-83062112021-07-25 Lipofection of Single Guide RNA Targeting MMP8 Decreases Proliferation and Migration in Lung Adenocarcinoma Cells Maradiaga, Oscar David Hernandez Mok, Pooi Ling Sivapragasam, Gothai Samrot, Antony V. Ali Khan, Mohammed Safwan Farhana, Aisha Alzahrani, Badr Tong, Jiabei Karuppiah, Thilakavathy Joseph, Narcisse M. S. Subbiah, Suresh Kumar Medicina (Kaunas) Article Background and Objectives: Matrix metalloproteinases (MMP) have been implicated as major determinants of tumour growth and metastasis, which are considered two of the main hallmarks of cancer. The interaction of MMP8 and other signalling molecules within and adjacent tumoral tissues, including immune cells, are rather elusive, particularly of adenocarcinoma cell type. In this study, we aimed to investigate the role of MMP8 in non-small cell lung cancer proliferation and invasiveness potential. Materials and Methods: We individually lipofected with two different single guide RNA (sgRNAs) that specifically targeted on MMP8, with CRISPR-Cas 9 protein into the cells. Results: Our results clearly indicated that the lipofection of these complexes could lead to reduced ability of A549 cells to survive and proliferate to form colonies. In addition, when compared to non-transfected cells, the experimental cell groups receiving sgRNAs demonstrated relatively decreased migration rate, hence, wider wound gaps in scratch assay. The quantitative real time-polymerase chain reaction (qRT-PCR) demonstrated significant reduction in the MAP-K, survivin and PI3-K gene expression. MMP8 might have protective roles over tumour growth and spread in our body. Conclusions: The delivery of sgRNAs targeting on the MMP8 gene could induce tumour cell death and arrest cell migratory activity. MDPI 2021-07-14 /pmc/articles/PMC8306211/ /pubmed/34356991 http://dx.doi.org/10.3390/medicina57070710 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Maradiaga, Oscar David Hernandez
Mok, Pooi Ling
Sivapragasam, Gothai
Samrot, Antony V.
Ali Khan, Mohammed Safwan
Farhana, Aisha
Alzahrani, Badr
Tong, Jiabei
Karuppiah, Thilakavathy
Joseph, Narcisse M. S.
Subbiah, Suresh Kumar
Lipofection of Single Guide RNA Targeting MMP8 Decreases Proliferation and Migration in Lung Adenocarcinoma Cells
title Lipofection of Single Guide RNA Targeting MMP8 Decreases Proliferation and Migration in Lung Adenocarcinoma Cells
title_full Lipofection of Single Guide RNA Targeting MMP8 Decreases Proliferation and Migration in Lung Adenocarcinoma Cells
title_fullStr Lipofection of Single Guide RNA Targeting MMP8 Decreases Proliferation and Migration in Lung Adenocarcinoma Cells
title_full_unstemmed Lipofection of Single Guide RNA Targeting MMP8 Decreases Proliferation and Migration in Lung Adenocarcinoma Cells
title_short Lipofection of Single Guide RNA Targeting MMP8 Decreases Proliferation and Migration in Lung Adenocarcinoma Cells
title_sort lipofection of single guide rna targeting mmp8 decreases proliferation and migration in lung adenocarcinoma cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8306211/
https://www.ncbi.nlm.nih.gov/pubmed/34356991
http://dx.doi.org/10.3390/medicina57070710
work_keys_str_mv AT maradiagaoscardavidhernandez lipofectionofsingleguidernatargetingmmp8decreasesproliferationandmigrationinlungadenocarcinomacells
AT mokpooiling lipofectionofsingleguidernatargetingmmp8decreasesproliferationandmigrationinlungadenocarcinomacells
AT sivapragasamgothai lipofectionofsingleguidernatargetingmmp8decreasesproliferationandmigrationinlungadenocarcinomacells
AT samrotantonyv lipofectionofsingleguidernatargetingmmp8decreasesproliferationandmigrationinlungadenocarcinomacells
AT alikhanmohammedsafwan lipofectionofsingleguidernatargetingmmp8decreasesproliferationandmigrationinlungadenocarcinomacells
AT farhanaaisha lipofectionofsingleguidernatargetingmmp8decreasesproliferationandmigrationinlungadenocarcinomacells
AT alzahranibadr lipofectionofsingleguidernatargetingmmp8decreasesproliferationandmigrationinlungadenocarcinomacells
AT tongjiabei lipofectionofsingleguidernatargetingmmp8decreasesproliferationandmigrationinlungadenocarcinomacells
AT karuppiahthilakavathy lipofectionofsingleguidernatargetingmmp8decreasesproliferationandmigrationinlungadenocarcinomacells
AT josephnarcissems lipofectionofsingleguidernatargetingmmp8decreasesproliferationandmigrationinlungadenocarcinomacells
AT subbiahsureshkumar lipofectionofsingleguidernatargetingmmp8decreasesproliferationandmigrationinlungadenocarcinomacells