Role of 3-Mercaptopyruvate Sulfurtransferase in the Regulation of Proliferation, Migration, and Bioenergetics in Murine Colon Cancer Cells
3-mercaptopyruvate sulfurtransferase (3-MST) has emerged as one of the significant sources of biologically active sulfur species in various mammalian cells. The current study was designed to investigate the functional role of 3-MST’s catalytic activity in the murine colon cancer cell line...
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| Format: | Article |
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MDPI AG
2020-03-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/10/3/447 |
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| author | Fiona Augsburger Elisa B. Randi Mathieu Jendly Kelly Ascencao Nahzli Dilek Csaba Szabo |
| author_facet | Fiona Augsburger Elisa B. Randi Mathieu Jendly Kelly Ascencao Nahzli Dilek Csaba Szabo |
| author_sort | Fiona Augsburger |
| collection | DOAJ |
| description | 3-mercaptopyruvate sulfurtransferase (3-MST) has emerged as one of the significant sources of biologically active sulfur species in various mammalian cells. The current study was designed to investigate the functional role of 3-MST’s catalytic activity in the murine colon cancer cell line CT26. The novel pharmacological 3-MST inhibitor HMPSNE was used to assess cancer cell proliferation, migration and bioenergetics in vitro. Methods included measurements of cell viability (MTT and LDH assays), cell proliferation and in vitro wound healing (IncuCyte) and cellular bioenergetics (Seahorse extracellular flux analysis). 3-MST expression was detected by Western blotting; H<sub>2</sub>S production was measured by the fluorescent dye AzMC. The results show that CT26 cells express 3-MST protein and mRNA, as well as several enzymes involved in H<sub>2</sub>S degradation (TST, ETHE1). Pharmacological inhibition of 3-MST concentration-dependently suppressed H<sub>2</sub>S production and, at 100 and 300 µM, attenuated CT26 proliferation and migration. HMPSNE exerted a bell-shaped effect on several cellular bioenergetic parameters related to oxidative phosphorylation, while other bioenergetic parameters were either unaffected or inhibited at the highest concentration of the inhibitor tested (300 µM). In contrast to 3-MST, the expression of CBS (another H<sub>2</sub>S producing enzyme which has been previously implicated in the regulation of various biological parameters in other tumor cells) was not detectable in CT26 cells and pharmacological inhibition of CBS exerted no significant effects on CT26 proliferation or bioenergetics. In summary, 3-MST catalytic activity significantly contributes to the regulation of cellular proliferation, migration and bioenergetics in CT26 murine colon cancer cells. The current studies identify 3-MST as the principal source of biologically active H<sub>2</sub>S in this cell line. |
| first_indexed | 2024-04-14T06:26:03Z |
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| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-04-14T06:26:03Z |
| publishDate | 2020-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-4f2941cc022b455cae9aeb773d17ceb12022-12-22T02:07:47ZengMDPI AGBiomolecules2218-273X2020-03-0110344710.3390/biom10030447biom10030447Role of 3-Mercaptopyruvate Sulfurtransferase in the Regulation of Proliferation, Migration, and Bioenergetics in Murine Colon Cancer CellsFiona Augsburger0Elisa B. Randi1Mathieu Jendly2Kelly Ascencao3Nahzli Dilek4Csaba Szabo5Chair of Pharmacology, Section of Medicine, University of Fribourg, 1700 Fribourg, SwitzerlandChair of Pharmacology, Section of Medicine, University of Fribourg, 1700 Fribourg, SwitzerlandChair of Pharmacology, Section of Medicine, University of Fribourg, 1700 Fribourg, SwitzerlandChair of Pharmacology, Section of Medicine, University of Fribourg, 1700 Fribourg, SwitzerlandChair of Pharmacology, Section of Medicine, University of Fribourg, 1700 Fribourg, SwitzerlandChair of Pharmacology, Section of Medicine, University of Fribourg, 1700 Fribourg, Switzerland3-mercaptopyruvate sulfurtransferase (3-MST) has emerged as one of the significant sources of biologically active sulfur species in various mammalian cells. The current study was designed to investigate the functional role of 3-MST’s catalytic activity in the murine colon cancer cell line CT26. The novel pharmacological 3-MST inhibitor HMPSNE was used to assess cancer cell proliferation, migration and bioenergetics in vitro. Methods included measurements of cell viability (MTT and LDH assays), cell proliferation and in vitro wound healing (IncuCyte) and cellular bioenergetics (Seahorse extracellular flux analysis). 3-MST expression was detected by Western blotting; H<sub>2</sub>S production was measured by the fluorescent dye AzMC. The results show that CT26 cells express 3-MST protein and mRNA, as well as several enzymes involved in H<sub>2</sub>S degradation (TST, ETHE1). Pharmacological inhibition of 3-MST concentration-dependently suppressed H<sub>2</sub>S production and, at 100 and 300 µM, attenuated CT26 proliferation and migration. HMPSNE exerted a bell-shaped effect on several cellular bioenergetic parameters related to oxidative phosphorylation, while other bioenergetic parameters were either unaffected or inhibited at the highest concentration of the inhibitor tested (300 µM). In contrast to 3-MST, the expression of CBS (another H<sub>2</sub>S producing enzyme which has been previously implicated in the regulation of various biological parameters in other tumor cells) was not detectable in CT26 cells and pharmacological inhibition of CBS exerted no significant effects on CT26 proliferation or bioenergetics. In summary, 3-MST catalytic activity significantly contributes to the regulation of cellular proliferation, migration and bioenergetics in CT26 murine colon cancer cells. The current studies identify 3-MST as the principal source of biologically active H<sub>2</sub>S in this cell line.https://www.mdpi.com/2218-273X/10/3/447hydrogen sulfidebioenergeticsgasotransmittersnitric oxideproliferationmigrationmitochondria |
| spellingShingle | Fiona Augsburger Elisa B. Randi Mathieu Jendly Kelly Ascencao Nahzli Dilek Csaba Szabo Role of 3-Mercaptopyruvate Sulfurtransferase in the Regulation of Proliferation, Migration, and Bioenergetics in Murine Colon Cancer Cells Biomolecules hydrogen sulfide bioenergetics gasotransmitters nitric oxide proliferation migration mitochondria |
| title | Role of 3-Mercaptopyruvate Sulfurtransferase in the Regulation of Proliferation, Migration, and Bioenergetics in Murine Colon Cancer Cells |
| title_full | Role of 3-Mercaptopyruvate Sulfurtransferase in the Regulation of Proliferation, Migration, and Bioenergetics in Murine Colon Cancer Cells |
| title_fullStr | Role of 3-Mercaptopyruvate Sulfurtransferase in the Regulation of Proliferation, Migration, and Bioenergetics in Murine Colon Cancer Cells |
| title_full_unstemmed | Role of 3-Mercaptopyruvate Sulfurtransferase in the Regulation of Proliferation, Migration, and Bioenergetics in Murine Colon Cancer Cells |
| title_short | Role of 3-Mercaptopyruvate Sulfurtransferase in the Regulation of Proliferation, Migration, and Bioenergetics in Murine Colon Cancer Cells |
| title_sort | role of 3 mercaptopyruvate sulfurtransferase in the regulation of proliferation migration and bioenergetics in murine colon cancer cells |
| topic | hydrogen sulfide bioenergetics gasotransmitters nitric oxide proliferation migration mitochondria |
| url | https://www.mdpi.com/2218-273X/10/3/447 |
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