The Adaptation of MCF-7 Breast Cancer Spheroids to the Chemotherapeutic Doxorubicin: The Dynamic Role of Phase I Drug Metabolizing Enzymes

<b>Background/Objectives:</b> Drug resistance (DR) is a major challenge in cancer therapy, contributing to approximately 90% of cancer-related deaths. While alterations in drug metabolism are known to be key drivers of DR, their role—particularly in the early stages of acquired chemoresi...

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Main Authors: Daniel Crispim, Carolina Ramos, Francisco Esteves, Michel Kranendonk
Format: Article
Language:English
Published: MDPI AG 2025-02-01
Series:Metabolites
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Online Access:https://www.mdpi.com/2218-1989/15/2/136
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author Daniel Crispim
Carolina Ramos
Francisco Esteves
Michel Kranendonk
author_facet Daniel Crispim
Carolina Ramos
Francisco Esteves
Michel Kranendonk
author_sort Daniel Crispim
collection DOAJ
description <b>Background/Objectives:</b> Drug resistance (DR) is a major challenge in cancer therapy, contributing to approximately 90% of cancer-related deaths. While alterations in drug metabolism are known to be key drivers of DR, their role—particularly in the early stages of acquired chemoresistance—remains understudied. Phase I drug-metabolizing enzymes (DMEs), especially cytochrome P450s (CYPs), significantly influence the metabolic fate of chemotherapeutic agents, directly affecting drug response. This study aimed to investigate the role of Phase I DMEs in the early metabolic adaptation of breast cancer (BC) MCF-7 cells to doxorubicin (DOX). <b>Methods:</b> Four types of spheroids were generated from MCF-7 cells that were either DOX-sensitive (DOX<sup>S</sup>) or adapted to low concentrations of the chemotherapeutic agent (DOX<sup>A</sup> 25, 35, and 45 nM). The expression levels of 92 Phase I DMEs and the activities of specific CYP isoforms were assessed in both DOX<sup>S</sup> and DOX<sup>A</sup> spheroids. <b>Results:</b> A total of twenty-four DMEs, including fifteen CYPs and nine oxidoreductases, were found to be differentially expressed in DOX<sup>A</sup> spheroids. Pathway analysis identified key roles for the differentially expressed DMEs in physiologically relevant pathways, including the metabolism of drugs, arachidonic acid, retinoic acid, and vitamin D. <b>Conclusions:</b> The deconvolution of these pathways highlights a highly dynamic process driving early-stage DOX resistance, with a prominent role of CYP3A-dependent metabolism in DOX adaptation. Our findings provide valuable insights into the underlying molecular mechanisms driving the early adaptation of MCF-7 cells to DOX exposure.
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spelling doaj-art-ffb5ecc09eb649e4ae6d048a530b16982025-08-20T03:12:04ZengMDPI AGMetabolites2218-19892025-02-0115213610.3390/metabo15020136The Adaptation of MCF-7 Breast Cancer Spheroids to the Chemotherapeutic Doxorubicin: The Dynamic Role of Phase I Drug Metabolizing EnzymesDaniel Crispim0Carolina Ramos1Francisco Esteves2Michel Kranendonk3Comprehensive Health Research Centre (CHRC) NOVA Medical School | Faculty of Medical Sciences, Universidade NOVA de Lisboa, 1169-056 Lisboa, PortugalComprehensive Health Research Centre (CHRC) NOVA Medical School | Faculty of Medical Sciences, Universidade NOVA de Lisboa, 1169-056 Lisboa, PortugalComprehensive Health Research Centre (CHRC) NOVA Medical School | Faculty of Medical Sciences, Universidade NOVA de Lisboa, 1169-056 Lisboa, PortugalComprehensive Health Research Centre (CHRC) NOVA Medical School | Faculty of Medical Sciences, Universidade NOVA de Lisboa, 1169-056 Lisboa, Portugal<b>Background/Objectives:</b> Drug resistance (DR) is a major challenge in cancer therapy, contributing to approximately 90% of cancer-related deaths. While alterations in drug metabolism are known to be key drivers of DR, their role—particularly in the early stages of acquired chemoresistance—remains understudied. Phase I drug-metabolizing enzymes (DMEs), especially cytochrome P450s (CYPs), significantly influence the metabolic fate of chemotherapeutic agents, directly affecting drug response. This study aimed to investigate the role of Phase I DMEs in the early metabolic adaptation of breast cancer (BC) MCF-7 cells to doxorubicin (DOX). <b>Methods:</b> Four types of spheroids were generated from MCF-7 cells that were either DOX-sensitive (DOX<sup>S</sup>) or adapted to low concentrations of the chemotherapeutic agent (DOX<sup>A</sup> 25, 35, and 45 nM). The expression levels of 92 Phase I DMEs and the activities of specific CYP isoforms were assessed in both DOX<sup>S</sup> and DOX<sup>A</sup> spheroids. <b>Results:</b> A total of twenty-four DMEs, including fifteen CYPs and nine oxidoreductases, were found to be differentially expressed in DOX<sup>A</sup> spheroids. Pathway analysis identified key roles for the differentially expressed DMEs in physiologically relevant pathways, including the metabolism of drugs, arachidonic acid, retinoic acid, and vitamin D. <b>Conclusions:</b> The deconvolution of these pathways highlights a highly dynamic process driving early-stage DOX resistance, with a prominent role of CYP3A-dependent metabolism in DOX adaptation. Our findings provide valuable insights into the underlying molecular mechanisms driving the early adaptation of MCF-7 cells to DOX exposure.https://www.mdpi.com/2218-1989/15/2/136breast cancer (BC)drug-metabolizing enzymes (DMEs)3D models/spheroidsdoxorubicin (DOX)cytochrome P450s (CYPs)oxidoreductases
spellingShingle Daniel Crispim
Carolina Ramos
Francisco Esteves
Michel Kranendonk
The Adaptation of MCF-7 Breast Cancer Spheroids to the Chemotherapeutic Doxorubicin: The Dynamic Role of Phase I Drug Metabolizing Enzymes
Metabolites
breast cancer (BC)
drug-metabolizing enzymes (DMEs)
3D models/spheroids
doxorubicin (DOX)
cytochrome P450s (CYPs)
oxidoreductases
title The Adaptation of MCF-7 Breast Cancer Spheroids to the Chemotherapeutic Doxorubicin: The Dynamic Role of Phase I Drug Metabolizing Enzymes
title_full The Adaptation of MCF-7 Breast Cancer Spheroids to the Chemotherapeutic Doxorubicin: The Dynamic Role of Phase I Drug Metabolizing Enzymes
title_fullStr The Adaptation of MCF-7 Breast Cancer Spheroids to the Chemotherapeutic Doxorubicin: The Dynamic Role of Phase I Drug Metabolizing Enzymes
title_full_unstemmed The Adaptation of MCF-7 Breast Cancer Spheroids to the Chemotherapeutic Doxorubicin: The Dynamic Role of Phase I Drug Metabolizing Enzymes
title_short The Adaptation of MCF-7 Breast Cancer Spheroids to the Chemotherapeutic Doxorubicin: The Dynamic Role of Phase I Drug Metabolizing Enzymes
title_sort adaptation of mcf 7 breast cancer spheroids to the chemotherapeutic doxorubicin the dynamic role of phase i drug metabolizing enzymes
topic breast cancer (BC)
drug-metabolizing enzymes (DMEs)
3D models/spheroids
doxorubicin (DOX)
cytochrome P450s (CYPs)
oxidoreductases
url https://www.mdpi.com/2218-1989/15/2/136
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