A Role for H<sub>2</sub>O<sub>2</sub> and TRPM2 in the Induction of Cell Death: Studies in KGN Cells
Recent studies showed that KGN cells, derived from a human granulosa cell tumor (GCT), express NADPH oxidase 4 (NOX4), an important source of H<sub>2</sub>O<sub>2</sub>. Transient receptor potential melastatin 2 (TRPM2) channel is a Ca<sup>2+</sup> permeable catio...
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| Format: | Article |
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MDPI AG
2019-10-01
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| Series: | Antioxidants |
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| Online Access: | https://www.mdpi.com/2076-3921/8/11/518 |
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| author | Carsten Theo Hack Theresa Buck Konstantin Bagnjuk Katja Eubler Lars Kunz Doris Mayr Artur Mayerhofer |
| author_facet | Carsten Theo Hack Theresa Buck Konstantin Bagnjuk Katja Eubler Lars Kunz Doris Mayr Artur Mayerhofer |
| author_sort | Carsten Theo Hack |
| collection | DOAJ |
| description | Recent studies showed that KGN cells, derived from a human granulosa cell tumor (GCT), express NADPH oxidase 4 (NOX4), an important source of H<sub>2</sub>O<sub>2</sub>. Transient receptor potential melastatin 2 (TRPM2) channel is a Ca<sup>2+</sup> permeable cation channel that can be activated by H<sub>2</sub>O<sub>2</sub> and plays an important role in cellular functions. It is also able to promote susceptibility to cell death. We studied expression and functionality of TRPM2 in KGN cells and examined GCT tissue microarrays (TMAs) to explore in vivo relevance. We employed live cell, calcium and mitochondrial imaging, viability assays, fluorescence activated cell sorting (FACS) analysis, Western blotting and immunohistochemistry. We confirmed that KGN cells produce H<sub>2</sub>O<sub>2</sub> and found that they express functional TRPM2. H<sub>2</sub>O<sub>2</sub> increased intracellular Ca<sup>2+</sup> levels and N-(p-Amylcinnamoyl)anthranilic acid (ACA), a TRPM2 inhibitor, blocked this action. H<sub>2</sub>O<sub>2</sub> caused mitochondrial fragmentation and apoptotic cell death, which could be attenuated by a scavenger (Trolox). Immunohistochemistry showed parallel expression of NOX4 and TRPM2 in all 73 tumor samples examined. The results suggest that GCTs can be endowed with a system that may convey susceptibility to cell death. If so, induction of oxidative stress may be beneficial in GCT therapy. Our results also imply a therapeutic potential for TRPM2 as a drug target in GCTs. |
| first_indexed | 2024-03-12T19:09:49Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 2076-3921 |
| language | English |
| last_indexed | 2024-03-12T19:09:49Z |
| publishDate | 2019-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antioxidants |
| spelling | doaj.art-178f3648638a463d9089de8901590e2c2023-08-02T05:58:37ZengMDPI AGAntioxidants2076-39212019-10-0181151810.3390/antiox8110518antiox8110518A Role for H<sub>2</sub>O<sub>2</sub> and TRPM2 in the Induction of Cell Death: Studies in KGN CellsCarsten Theo Hack0Theresa Buck1Konstantin Bagnjuk2Katja Eubler3Lars Kunz4Doris Mayr5Artur Mayerhofer6Biomedical Center Munich (BMC), Cell Biology, Anatomy III, Ludwig-Maximilian-University (LMU), D-82152 Planegg, GermanyBiomedical Center Munich (BMC), Cell Biology, Anatomy III, Ludwig-Maximilian-University (LMU), D-82152 Planegg, GermanyBiomedical Center Munich (BMC), Cell Biology, Anatomy III, Ludwig-Maximilian-University (LMU), D-82152 Planegg, GermanyBiomedical Center Munich (BMC), Cell Biology, Anatomy III, Ludwig-Maximilian-University (LMU), D-82152 Planegg, GermanyDivision of Neurobiology, Department Biology II, Ludwig-Maximilian-University (LMU), D-82152 Planegg, GermanyInstitute of Pathology, Ludwig-Maximilian-University (LMU), D-80337 München, GermanyBiomedical Center Munich (BMC), Cell Biology, Anatomy III, Ludwig-Maximilian-University (LMU), D-82152 Planegg, GermanyRecent studies showed that KGN cells, derived from a human granulosa cell tumor (GCT), express NADPH oxidase 4 (NOX4), an important source of H<sub>2</sub>O<sub>2</sub>. Transient receptor potential melastatin 2 (TRPM2) channel is a Ca<sup>2+</sup> permeable cation channel that can be activated by H<sub>2</sub>O<sub>2</sub> and plays an important role in cellular functions. It is also able to promote susceptibility to cell death. We studied expression and functionality of TRPM2 in KGN cells and examined GCT tissue microarrays (TMAs) to explore in vivo relevance. We employed live cell, calcium and mitochondrial imaging, viability assays, fluorescence activated cell sorting (FACS) analysis, Western blotting and immunohistochemistry. We confirmed that KGN cells produce H<sub>2</sub>O<sub>2</sub> and found that they express functional TRPM2. H<sub>2</sub>O<sub>2</sub> increased intracellular Ca<sup>2+</sup> levels and N-(p-Amylcinnamoyl)anthranilic acid (ACA), a TRPM2 inhibitor, blocked this action. H<sub>2</sub>O<sub>2</sub> caused mitochondrial fragmentation and apoptotic cell death, which could be attenuated by a scavenger (Trolox). Immunohistochemistry showed parallel expression of NOX4 and TRPM2 in all 73 tumor samples examined. The results suggest that GCTs can be endowed with a system that may convey susceptibility to cell death. If so, induction of oxidative stress may be beneficial in GCT therapy. Our results also imply a therapeutic potential for TRPM2 as a drug target in GCTs.https://www.mdpi.com/2076-3921/8/11/518ovarycalcium channeltroloxgranulosa cell tumorcell deathmitochondria |
| spellingShingle | Carsten Theo Hack Theresa Buck Konstantin Bagnjuk Katja Eubler Lars Kunz Doris Mayr Artur Mayerhofer A Role for H<sub>2</sub>O<sub>2</sub> and TRPM2 in the Induction of Cell Death: Studies in KGN Cells Antioxidants ovary calcium channel trolox granulosa cell tumor cell death mitochondria |
| title | A Role for H<sub>2</sub>O<sub>2</sub> and TRPM2 in the Induction of Cell Death: Studies in KGN Cells |
| title_full | A Role for H<sub>2</sub>O<sub>2</sub> and TRPM2 in the Induction of Cell Death: Studies in KGN Cells |
| title_fullStr | A Role for H<sub>2</sub>O<sub>2</sub> and TRPM2 in the Induction of Cell Death: Studies in KGN Cells |
| title_full_unstemmed | A Role for H<sub>2</sub>O<sub>2</sub> and TRPM2 in the Induction of Cell Death: Studies in KGN Cells |
| title_short | A Role for H<sub>2</sub>O<sub>2</sub> and TRPM2 in the Induction of Cell Death: Studies in KGN Cells |
| title_sort | role for h sub 2 sub o sub 2 sub and trpm2 in the induction of cell death studies in kgn cells |
| topic | ovary calcium channel trolox granulosa cell tumor cell death mitochondria |
| url | https://www.mdpi.com/2076-3921/8/11/518 |
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