Cargando…
High Performance Liquid Chromatography–Tandem Mass Spectrometry Method for Correlating the Metabolic Changes of Lactate, Pyruvate and L-Glutamine with Induced Tamoxifen Resistant MCF-7 Cell Line Potential Molecular Changes
Breast cancer is one of the most prevalent cancers worldwide usually treated with Tamoxifen. Tamoxifen resistance development is the most challenging issue in an initially responsive breast tumor, and mechanisms of resistance are still under investigation. The objective of this study is to develop a...
Autores principales: | , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8399909/ https://www.ncbi.nlm.nih.gov/pubmed/34443413 http://dx.doi.org/10.3390/molecules26164824 |
_version_ | 1783745188066230272 |
---|---|
author | Alhusban, Ala A. Albustanji, Sokiyna Hamadneh, Lama A. Shallan, Aliaa I. |
author_facet | Alhusban, Ala A. Albustanji, Sokiyna Hamadneh, Lama A. Shallan, Aliaa I. |
author_sort | Alhusban, Ala A. |
collection | PubMed |
description | Breast cancer is one of the most prevalent cancers worldwide usually treated with Tamoxifen. Tamoxifen resistance development is the most challenging issue in an initially responsive breast tumor, and mechanisms of resistance are still under investigation. The objective of this study is to develop and validate a selective, sensitive, and simultaneous high performance liquid chromatography–tandem mass spectrometry method to explore the changes in substrates and metabolites in supernatant media of developed Tamoxifen resistance MCF-7 cells. We focus on the determination of lactate, pyruvate, and L-glutamine which enables the tracking of changes in metabolic pathways as a result of the resistance process. Chromatographic separation was achieved within 3.5 min. using a HILIC column (4.6 × 100 mm, 3.5 µm particle size) and mobile phase of 0.05 M acetic acid–ammonium acetate buffer solution pH 3.0: Acetonitrile (40:60 v/v). The linear range was 0.11–2.25, 0.012–0.227, and 0.02–0.20 mM for lactate, pyruvate, and L-glutamine, respectively. Within- and between-run accuracy was in the range 98.94-105.50% with precision (CV, %) of ≤0.86%. The results revealed a significant increase in both lactate and pyruvate production after acquiring the resistant. An increase in L-glutamine levels was also observed and could be attributed to its over production or decline in its consumption. Therefore, further tracking of genes responsible of lactate, pyruvate, and glutamine metabolic pathways should be performed in parallel to provide in-depth explanation of resistance mechanism. |
format | Online Article Text |
id | pubmed-8399909 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-83999092021-08-29 High Performance Liquid Chromatography–Tandem Mass Spectrometry Method for Correlating the Metabolic Changes of Lactate, Pyruvate and L-Glutamine with Induced Tamoxifen Resistant MCF-7 Cell Line Potential Molecular Changes Alhusban, Ala A. Albustanji, Sokiyna Hamadneh, Lama A. Shallan, Aliaa I. Molecules Article Breast cancer is one of the most prevalent cancers worldwide usually treated with Tamoxifen. Tamoxifen resistance development is the most challenging issue in an initially responsive breast tumor, and mechanisms of resistance are still under investigation. The objective of this study is to develop and validate a selective, sensitive, and simultaneous high performance liquid chromatography–tandem mass spectrometry method to explore the changes in substrates and metabolites in supernatant media of developed Tamoxifen resistance MCF-7 cells. We focus on the determination of lactate, pyruvate, and L-glutamine which enables the tracking of changes in metabolic pathways as a result of the resistance process. Chromatographic separation was achieved within 3.5 min. using a HILIC column (4.6 × 100 mm, 3.5 µm particle size) and mobile phase of 0.05 M acetic acid–ammonium acetate buffer solution pH 3.0: Acetonitrile (40:60 v/v). The linear range was 0.11–2.25, 0.012–0.227, and 0.02–0.20 mM for lactate, pyruvate, and L-glutamine, respectively. Within- and between-run accuracy was in the range 98.94-105.50% with precision (CV, %) of ≤0.86%. The results revealed a significant increase in both lactate and pyruvate production after acquiring the resistant. An increase in L-glutamine levels was also observed and could be attributed to its over production or decline in its consumption. Therefore, further tracking of genes responsible of lactate, pyruvate, and glutamine metabolic pathways should be performed in parallel to provide in-depth explanation of resistance mechanism. MDPI 2021-08-10 /pmc/articles/PMC8399909/ /pubmed/34443413 http://dx.doi.org/10.3390/molecules26164824 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 Alhusban, Ala A. Albustanji, Sokiyna Hamadneh, Lama A. Shallan, Aliaa I. High Performance Liquid Chromatography–Tandem Mass Spectrometry Method for Correlating the Metabolic Changes of Lactate, Pyruvate and L-Glutamine with Induced Tamoxifen Resistant MCF-7 Cell Line Potential Molecular Changes |
title | High Performance Liquid Chromatography–Tandem Mass Spectrometry Method for Correlating the Metabolic Changes of Lactate, Pyruvate and L-Glutamine with Induced Tamoxifen Resistant MCF-7 Cell Line Potential Molecular Changes |
title_full | High Performance Liquid Chromatography–Tandem Mass Spectrometry Method for Correlating the Metabolic Changes of Lactate, Pyruvate and L-Glutamine with Induced Tamoxifen Resistant MCF-7 Cell Line Potential Molecular Changes |
title_fullStr | High Performance Liquid Chromatography–Tandem Mass Spectrometry Method for Correlating the Metabolic Changes of Lactate, Pyruvate and L-Glutamine with Induced Tamoxifen Resistant MCF-7 Cell Line Potential Molecular Changes |
title_full_unstemmed | High Performance Liquid Chromatography–Tandem Mass Spectrometry Method for Correlating the Metabolic Changes of Lactate, Pyruvate and L-Glutamine with Induced Tamoxifen Resistant MCF-7 Cell Line Potential Molecular Changes |
title_short | High Performance Liquid Chromatography–Tandem Mass Spectrometry Method for Correlating the Metabolic Changes of Lactate, Pyruvate and L-Glutamine with Induced Tamoxifen Resistant MCF-7 Cell Line Potential Molecular Changes |
title_sort | high performance liquid chromatography–tandem mass spectrometry method for correlating the metabolic changes of lactate, pyruvate and l-glutamine with induced tamoxifen resistant mcf-7 cell line potential molecular changes |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8399909/ https://www.ncbi.nlm.nih.gov/pubmed/34443413 http://dx.doi.org/10.3390/molecules26164824 |
work_keys_str_mv | AT alhusbanalaa highperformanceliquidchromatographytandemmassspectrometrymethodforcorrelatingthemetabolicchangesoflactatepyruvateandlglutaminewithinducedtamoxifenresistantmcf7celllinepotentialmolecularchanges AT albustanjisokiyna highperformanceliquidchromatographytandemmassspectrometrymethodforcorrelatingthemetabolicchangesoflactatepyruvateandlglutaminewithinducedtamoxifenresistantmcf7celllinepotentialmolecularchanges AT hamadnehlamaa highperformanceliquidchromatographytandemmassspectrometrymethodforcorrelatingthemetabolicchangesoflactatepyruvateandlglutaminewithinducedtamoxifenresistantmcf7celllinepotentialmolecularchanges AT shallanaliaai highperformanceliquidchromatographytandemmassspectrometrymethodforcorrelatingthemetabolicchangesoflactatepyruvateandlglutaminewithinducedtamoxifenresistantmcf7celllinepotentialmolecularchanges |