Metabolomics Analysis Reveals Altered Metabolic Pathways and Response to Doxorubicin in Drug-Resistant Triple-Negative Breast Cancer Cells

Abstract

This study aimed to investigate metabolic changes following the acquisition of resistance to doxorubicin in the triple-negative breast cancer (TNBC) cell line MDA-MB-231. Two drug-resistant cell lines, DOX-RES-50 and DOX-RES-100, were generated by treating MDA-MB-231 cells with doxorubicin for 24 h and allowing them to recover for six weeks. Both drug-resistant cell lines demonstrated an increase in doxorubicin IC50 values, indicating acquired drug resistance. Metabolomics analysis showed clear separation between the parental MDA-MB-231 cell line and the drug-resistant cell lines. Pathway analysis revealed that arginine and proline metabolism, glutathione metabolism, and beta-alanine metabolism were significantly perturbed in the drug-resistant cell lines compared to the parental cell line. After matching signals to an in-house library of reference standards, significant decreases in short- and medium-chain acylcarnitines and significant increases in long-chain acylcarnitines, 5-oxoproline, and 7-ketodeoxycholic acid were observed in the resistant cell lines as compared to the parental MDA-MB-231 cell line. In addition to baseline metabolic differences, we also investigated differences in metabolic responses in resistant cell lines upon a second exposure at multiple concentrations. Results indicate that whereas the parental MDA-MB-231 cell line had many metabolites that responded to doxorubicin in a dose-dependent manner, the two resistant cell lines lost a dose-dependent response for the majority of these metabolites. The study’s findings provide insight into how metabolism is altered during the acquisition of resistance in TNBC cells and how the metabolic response to doxorubicin changes upon repeated treatment. This information can potentially identify novel targets to prevent or reverse multi-drug resistance in TNBC, and also demonstrate the usefulness of metabolomics technology in identifying new mechanisms of drug resistance in cancer and potential drug targets.

Unlocking the Molecular Secrets of Antifolate Drug Resistance: A Multi-Omics Investigation of the NCI-60 Cell Line Panel

“…Interestingly, the current study did not identify DHFR or TS as major differentiators of sensitive and resistant cells, but these have been primarily studied in acquired resistance settings. As our previous work has shown with alkylating agents, intrinsic and acquired resistance seem to be defined by distinct metabolic mechanisms, which may explain this difference [ 26 , 27 , 28 ].…”

MAPK14/p38α Shapes the Molecular Landscape of Endometrial Cancer and promotes Tumorigenic Characteristics

“…Peaks were matched to metabolites by retention time (RT ± 0.5 min, in-house matches only), exact mass (MS, <5 ppm), and MS/MS similarity (>30%). To communicate matching evidence an ontology system was used as described previously 122, 123 . Multivariate analysis was performed using SIMCA 16 (Sartorius Stedim Data Analytics AB, Umeå, Sweden) using the normalized, filtered data.…”

Cytotoxic, antioxidant, and cytoprotective properties of polyphenol‐enriched extracts from pecan nutshells in MDA‐MB‐231 breast cancer cells

“…The human MDA-MB-231 cell line used in this study allowed us to extend the research in breast cancer and is sensitive to chemotherapeutic drugs currently used for treatment this cancer, such as doxorubicin HCl. 10,19,20,34,35 The cytotoxicity of PCEE (10-200 mg/L) and doxorubicin HCl According to the US NCI plant screening program, a crude extract is generally considered to have in vitro cytotoxic activity if the IC 50 value (concentration that causes a reduction in cell viability to 50%) is less than 30 mg/L. 38 Thus, the PCEE evaluated in this study could be a source of antitumoral therapeutic agents against breast cancer.…”

“…One of these is the search for natural compounds with chemo-preventive or anticancer properties that could be used in combination with doxorubicin. [16][17][18][19][20] In the present work, we aimed to study the antitumor and the antioxidant effects of a purified phenolic compound-enriched extract (PCEE) on breast cancer and normal cell lines (MDA-MB-231 and CHO-K1, respectively) by using different in vitro assays and doxorubicin as a control.…”