Proteomic and Bioinformatic Investigation of Altered Pathways in Neuroglobin-Deficient Breast Cancer Cells
Neuroglobin (NGB) is a myoglobin-like monomeric globin that is involved in several processes, displaying a pivotal redox-dependent protective role in neuronal and extra-neuronal cells. NGB remarkably exerts its function upon upregulation by NGB inducers, such as 17β-estradiol (E2) and H<sub>2&...
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MDPI AG
2021-04-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/26/8/2397 |
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| author | Michele Costanzo Marco Fiocchetti Paolo Ascenzi Maria Marino Marianna Caterino Margherita Ruoppolo |
| author_facet | Michele Costanzo Marco Fiocchetti Paolo Ascenzi Maria Marino Marianna Caterino Margherita Ruoppolo |
| author_sort | Michele Costanzo |
| collection | DOAJ |
| description | Neuroglobin (NGB) is a myoglobin-like monomeric globin that is involved in several processes, displaying a pivotal redox-dependent protective role in neuronal and extra-neuronal cells. NGB remarkably exerts its function upon upregulation by NGB inducers, such as 17β-estradiol (E2) and H<sub>2</sub>O<sub>2</sub>. However, the molecular bases of NGB’s functions remain undefined, mainly in non-neuronal cancer cells. Human MCF-7 breast cancer cells with a knocked-out (KO) <i>NGB</i> gene obtained using CRISPR/Cas9 technology were analyzed using shotgun label-free quantitative proteomics in comparison with control cells. The differential proteomics experiments were also performed after treatment with E2, H<sub>2</sub>O<sub>2</sub>, and E2 + H<sub>2</sub>O<sub>2</sub>. All the runs acquired using liquid chromatography–tandem mass spectrometry were elaborated within the same MaxQuant analysis, leading to the quantification of 1872 proteins in the global proteomic dataset. Then, a differentially regulated protein dataset was obtained for each specific treatment. After the proteomic study, multiple bioinformatics analyses were performed to highlight unbalanced pathways and processes. Here, we report the proteomic and bioinformatic investigations concerning the effects on cellular processes of NGB deficiency and cell treatments. Globally, the main processes that were affected were related to the response to stress, cytoskeleton dynamics, apoptosis, and mitochondria-driven pathways. |
| first_indexed | 2024-03-10T12:08:58Z |
| format | Article |
| id | doaj.art-ba72553199f74ecba52f2d9ab4b2d604 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T12:08:58Z |
| publishDate | 2021-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-ba72553199f74ecba52f2d9ab4b2d6042023-11-21T16:23:01ZengMDPI AGMolecules1420-30492021-04-01268239710.3390/molecules26082397Proteomic and Bioinformatic Investigation of Altered Pathways in Neuroglobin-Deficient Breast Cancer CellsMichele Costanzo0Marco Fiocchetti1Paolo Ascenzi2Maria Marino3Marianna Caterino4Margherita Ruoppolo5Department of Molecular Medicine and Medical Biotechnology, School of Medicine, University of Naples Federico II, 80131 Naples, ItalyDepartment of Science, University Roma Tre, 00146 Rome, ItalyDepartment of Science, University Roma Tre, 00146 Rome, ItalyDepartment of Science, University Roma Tre, 00146 Rome, ItalyDepartment of Molecular Medicine and Medical Biotechnology, School of Medicine, University of Naples Federico II, 80131 Naples, ItalyDepartment of Molecular Medicine and Medical Biotechnology, School of Medicine, University of Naples Federico II, 80131 Naples, ItalyNeuroglobin (NGB) is a myoglobin-like monomeric globin that is involved in several processes, displaying a pivotal redox-dependent protective role in neuronal and extra-neuronal cells. NGB remarkably exerts its function upon upregulation by NGB inducers, such as 17β-estradiol (E2) and H<sub>2</sub>O<sub>2</sub>. However, the molecular bases of NGB’s functions remain undefined, mainly in non-neuronal cancer cells. Human MCF-7 breast cancer cells with a knocked-out (KO) <i>NGB</i> gene obtained using CRISPR/Cas9 technology were analyzed using shotgun label-free quantitative proteomics in comparison with control cells. The differential proteomics experiments were also performed after treatment with E2, H<sub>2</sub>O<sub>2</sub>, and E2 + H<sub>2</sub>O<sub>2</sub>. All the runs acquired using liquid chromatography–tandem mass spectrometry were elaborated within the same MaxQuant analysis, leading to the quantification of 1872 proteins in the global proteomic dataset. Then, a differentially regulated protein dataset was obtained for each specific treatment. After the proteomic study, multiple bioinformatics analyses were performed to highlight unbalanced pathways and processes. Here, we report the proteomic and bioinformatic investigations concerning the effects on cellular processes of NGB deficiency and cell treatments. Globally, the main processes that were affected were related to the response to stress, cytoskeleton dynamics, apoptosis, and mitochondria-driven pathways.https://www.mdpi.com/1420-3049/26/8/2397neuroglobin17β-estradiollabel-free proteomicsbioinformaticspathway analysisoxidative stress |
| spellingShingle | Michele Costanzo Marco Fiocchetti Paolo Ascenzi Maria Marino Marianna Caterino Margherita Ruoppolo Proteomic and Bioinformatic Investigation of Altered Pathways in Neuroglobin-Deficient Breast Cancer Cells Molecules neuroglobin 17β-estradiol label-free proteomics bioinformatics pathway analysis oxidative stress |
| title | Proteomic and Bioinformatic Investigation of Altered Pathways in Neuroglobin-Deficient Breast Cancer Cells |
| title_full | Proteomic and Bioinformatic Investigation of Altered Pathways in Neuroglobin-Deficient Breast Cancer Cells |
| title_fullStr | Proteomic and Bioinformatic Investigation of Altered Pathways in Neuroglobin-Deficient Breast Cancer Cells |
| title_full_unstemmed | Proteomic and Bioinformatic Investigation of Altered Pathways in Neuroglobin-Deficient Breast Cancer Cells |
| title_short | Proteomic and Bioinformatic Investigation of Altered Pathways in Neuroglobin-Deficient Breast Cancer Cells |
| title_sort | proteomic and bioinformatic investigation of altered pathways in neuroglobin deficient breast cancer cells |
| topic | neuroglobin 17β-estradiol label-free proteomics bioinformatics pathway analysis oxidative stress |
| url | https://www.mdpi.com/1420-3049/26/8/2397 |
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