Gene and Protein Expression Is Altered by Ascorbate Availability in Murine Macrophages Cultured under Tumour-Like Conditions
Tumour-associated macrophages (TAMs) are ubiquitously present in tumours and commonly associated with poor prognosis. In immune cells, ascorbate affects epigenetic regulation, differentiation and phenotype via its co-factor activity for the 2-oxoglutarate dependent dioxygenase enzymes. Here, we dete...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-03-01
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| Series: | Antioxidants |
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| Online Access: | https://www.mdpi.com/2076-3921/10/3/430 |
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| author | Abel D. Ang Margreet C. M. Vissers Eleanor R. Burgess Margaret J. Currie Gabi U. Dachs |
| author_facet | Abel D. Ang Margreet C. M. Vissers Eleanor R. Burgess Margaret J. Currie Gabi U. Dachs |
| author_sort | Abel D. Ang |
| collection | DOAJ |
| description | Tumour-associated macrophages (TAMs) are ubiquitously present in tumours and commonly associated with poor prognosis. In immune cells, ascorbate affects epigenetic regulation, differentiation and phenotype via its co-factor activity for the 2-oxoglutarate dependent dioxygenase enzymes. Here, we determined the effect of ascorbate on TAM development in response to tumour microenvironmental cues. Naïve murine bone marrow monocytes were cultured with Lewis Lung Carcinoma conditioned media (LLCM) or macrophage colony-stimulating factor (MCSF) to encourage the development into tumour-associated macrophages. Cells were stimulated with hypoxia (1% O<sub>2</sub>), with or without ascorbate (500 µM) supplementation. Cells and media were harvested for gene, cell surface marker and protein analyses. LLCM supported bone marrow monocyte growth with >90% of cells staining CD11b<sup>+</sup>F4/80<sup>+</sup>, indicative of monocytes/macrophages. LLCM-grown cells showed increased expression of M2-like and TAM genes compared to MCSF-grown cells, which further increased with hypoxia. In LLCM-grown cells, ascorbate supplementation was associated with increased F4/80 cell surface expression, and altered gene expression and protein secretion. Our study shows that ascorbate modifies monocyte phenotype when grown under tumour microenvironmental conditions, but this was not clearly associated with either a pro- or anti-tumour phenotype, and reflects a complex and nuanced response of macrophages to ascorbate. Overall, ascorbate supplementation clearly has molecular consequences for TAMs, but functional and clinical consequences remain unknown. |
| first_indexed | 2024-03-10T13:20:00Z |
| format | Article |
| id | doaj.art-198dd28567054dac9734b28ac02b5e5a |
| institution | Directory Open Access Journal |
| issn | 2076-3921 |
| language | English |
| last_indexed | 2024-03-10T13:20:00Z |
| publishDate | 2021-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antioxidants |
| spelling | doaj.art-198dd28567054dac9734b28ac02b5e5a2023-11-21T10:05:18ZengMDPI AGAntioxidants2076-39212021-03-0110343010.3390/antiox10030430Gene and Protein Expression Is Altered by Ascorbate Availability in Murine Macrophages Cultured under Tumour-Like ConditionsAbel D. Ang0Margreet C. M. Vissers1Eleanor R. Burgess2Margaret J. Currie3Gabi U. Dachs4Mackenzie Cancer Research Group, Department of Pathology & Biomedical Science, University of Otago Christchurch, Christchurch 8140, New ZealandCentre for Free Radical Research, Department of Pathology & Biomedical Science, University of Otago Christchurch, Christchurch 8140, New ZealandMackenzie Cancer Research Group, Department of Pathology & Biomedical Science, University of Otago Christchurch, Christchurch 8140, New ZealandMackenzie Cancer Research Group, Department of Pathology & Biomedical Science, University of Otago Christchurch, Christchurch 8140, New ZealandMackenzie Cancer Research Group, Department of Pathology & Biomedical Science, University of Otago Christchurch, Christchurch 8140, New ZealandTumour-associated macrophages (TAMs) are ubiquitously present in tumours and commonly associated with poor prognosis. In immune cells, ascorbate affects epigenetic regulation, differentiation and phenotype via its co-factor activity for the 2-oxoglutarate dependent dioxygenase enzymes. Here, we determined the effect of ascorbate on TAM development in response to tumour microenvironmental cues. Naïve murine bone marrow monocytes were cultured with Lewis Lung Carcinoma conditioned media (LLCM) or macrophage colony-stimulating factor (MCSF) to encourage the development into tumour-associated macrophages. Cells were stimulated with hypoxia (1% O<sub>2</sub>), with or without ascorbate (500 µM) supplementation. Cells and media were harvested for gene, cell surface marker and protein analyses. LLCM supported bone marrow monocyte growth with >90% of cells staining CD11b<sup>+</sup>F4/80<sup>+</sup>, indicative of monocytes/macrophages. LLCM-grown cells showed increased expression of M2-like and TAM genes compared to MCSF-grown cells, which further increased with hypoxia. In LLCM-grown cells, ascorbate supplementation was associated with increased F4/80 cell surface expression, and altered gene expression and protein secretion. Our study shows that ascorbate modifies monocyte phenotype when grown under tumour microenvironmental conditions, but this was not clearly associated with either a pro- or anti-tumour phenotype, and reflects a complex and nuanced response of macrophages to ascorbate. Overall, ascorbate supplementation clearly has molecular consequences for TAMs, but functional and clinical consequences remain unknown.https://www.mdpi.com/2076-3921/10/3/430ascorbatetumourtumour-associated macrophagesgene expressionmicroenvironmenthypoxia |
| spellingShingle | Abel D. Ang Margreet C. M. Vissers Eleanor R. Burgess Margaret J. Currie Gabi U. Dachs Gene and Protein Expression Is Altered by Ascorbate Availability in Murine Macrophages Cultured under Tumour-Like Conditions Antioxidants ascorbate tumour tumour-associated macrophages gene expression microenvironment hypoxia |
| title | Gene and Protein Expression Is Altered by Ascorbate Availability in Murine Macrophages Cultured under Tumour-Like Conditions |
| title_full | Gene and Protein Expression Is Altered by Ascorbate Availability in Murine Macrophages Cultured under Tumour-Like Conditions |
| title_fullStr | Gene and Protein Expression Is Altered by Ascorbate Availability in Murine Macrophages Cultured under Tumour-Like Conditions |
| title_full_unstemmed | Gene and Protein Expression Is Altered by Ascorbate Availability in Murine Macrophages Cultured under Tumour-Like Conditions |
| title_short | Gene and Protein Expression Is Altered by Ascorbate Availability in Murine Macrophages Cultured under Tumour-Like Conditions |
| title_sort | gene and protein expression is altered by ascorbate availability in murine macrophages cultured under tumour like conditions |
| topic | ascorbate tumour tumour-associated macrophages gene expression microenvironment hypoxia |
| url | https://www.mdpi.com/2076-3921/10/3/430 |
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