NFATc1 Regulation of Dexamethasone-Induced <i>TGFB2</i> Expression Is Cell Cycle Dependent in Trabecular Meshwork Cells
Although elevated TGFβ2 levels appear to be a causative factor in glaucoma pathogenesis, little is known about how TGFβ2 expression is regulated in the trabecular meshwork (TM). Here, we investigated if activation of the cytokine regulator NFATc1 controlled transcription of TGFβ2 in human TM cells b...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-02-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/12/3/504 |
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| author | Mark S. Filla Jennifer A. Faralli Caleigh R. Dunn Haania Khan Donna M. Peters |
| author_facet | Mark S. Filla Jennifer A. Faralli Caleigh R. Dunn Haania Khan Donna M. Peters |
| author_sort | Mark S. Filla |
| collection | DOAJ |
| description | Although elevated TGFβ2 levels appear to be a causative factor in glaucoma pathogenesis, little is known about how TGFβ2 expression is regulated in the trabecular meshwork (TM). Here, we investigated if activation of the cytokine regulator NFATc1 controlled transcription of TGFβ2 in human TM cells by using dexamethasone (DEX) to induce NFATc1 activity. The study used both proliferating and cell cycle arrested quiescent cells. Cell cycle arrest was achieved by either cell–cell contact inhibition or serum starvation. β-catenin staining and p21 and Ki-67 nuclear labeling were used to verify the formation of cell–cell contacts and activity of the cell cycle. NFATc1 inhibitors cyclosporine A (CsA) or 11R-VIVIT were used to determine the role of NFATc1. mRNA levels were determined by RT-qPCR. DEX increased TGFβ2 mRNA expression by 3.5-fold in proliferating cells but not in quiescent cells or serum-starved cells, and both CsA and 11R-VIVIT inhibited this increase. In contrast, the expression of other DEX/NFATc1-induced mRNAs (myocilin and β3 integrin) occurred regardless of the proliferative state of the cells. These studies show that NAFTc1 regulates TGFβ2 transcription in TM cells and reveals a previously unknown connection between the TM cell cycle and modulation of gene expression by NFATc1 and/or DEX in TM cells. |
| first_indexed | 2024-03-11T09:48:56Z |
| format | Article |
| id | doaj.art-41d56406c9ce4999bd255fd8657921f7 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-11T09:48:56Z |
| publishDate | 2023-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-41d56406c9ce4999bd255fd8657921f72023-11-16T16:22:35ZengMDPI AGCells2073-44092023-02-0112350410.3390/cells12030504NFATc1 Regulation of Dexamethasone-Induced <i>TGFB2</i> Expression Is Cell Cycle Dependent in Trabecular Meshwork CellsMark S. Filla0Jennifer A. Faralli1Caleigh R. Dunn2Haania Khan3Donna M. Peters4Pathology & Laboratory Medicine, University of Wisconsin School of Medicine and Public Health, Madison, WI 53705, USAPathology & Laboratory Medicine, University of Wisconsin School of Medicine and Public Health, Madison, WI 53705, USAPathology & Laboratory Medicine, University of Wisconsin School of Medicine and Public Health, Madison, WI 53705, USAPathology & Laboratory Medicine, University of Wisconsin School of Medicine and Public Health, Madison, WI 53705, USAPathology & Laboratory Medicine, University of Wisconsin School of Medicine and Public Health, Madison, WI 53705, USAAlthough elevated TGFβ2 levels appear to be a causative factor in glaucoma pathogenesis, little is known about how TGFβ2 expression is regulated in the trabecular meshwork (TM). Here, we investigated if activation of the cytokine regulator NFATc1 controlled transcription of TGFβ2 in human TM cells by using dexamethasone (DEX) to induce NFATc1 activity. The study used both proliferating and cell cycle arrested quiescent cells. Cell cycle arrest was achieved by either cell–cell contact inhibition or serum starvation. β-catenin staining and p21 and Ki-67 nuclear labeling were used to verify the formation of cell–cell contacts and activity of the cell cycle. NFATc1 inhibitors cyclosporine A (CsA) or 11R-VIVIT were used to determine the role of NFATc1. mRNA levels were determined by RT-qPCR. DEX increased TGFβ2 mRNA expression by 3.5-fold in proliferating cells but not in quiescent cells or serum-starved cells, and both CsA and 11R-VIVIT inhibited this increase. In contrast, the expression of other DEX/NFATc1-induced mRNAs (myocilin and β3 integrin) occurred regardless of the proliferative state of the cells. These studies show that NAFTc1 regulates TGFβ2 transcription in TM cells and reveals a previously unknown connection between the TM cell cycle and modulation of gene expression by NFATc1 and/or DEX in TM cells.https://www.mdpi.com/2073-4409/12/3/504trabecular meshworkTGFβ2NFATc1dexamethasonecell cyclequiescence |
| spellingShingle | Mark S. Filla Jennifer A. Faralli Caleigh R. Dunn Haania Khan Donna M. Peters NFATc1 Regulation of Dexamethasone-Induced <i>TGFB2</i> Expression Is Cell Cycle Dependent in Trabecular Meshwork Cells Cells trabecular meshwork TGFβ2 NFATc1 dexamethasone cell cycle quiescence |
| title | NFATc1 Regulation of Dexamethasone-Induced <i>TGFB2</i> Expression Is Cell Cycle Dependent in Trabecular Meshwork Cells |
| title_full | NFATc1 Regulation of Dexamethasone-Induced <i>TGFB2</i> Expression Is Cell Cycle Dependent in Trabecular Meshwork Cells |
| title_fullStr | NFATc1 Regulation of Dexamethasone-Induced <i>TGFB2</i> Expression Is Cell Cycle Dependent in Trabecular Meshwork Cells |
| title_full_unstemmed | NFATc1 Regulation of Dexamethasone-Induced <i>TGFB2</i> Expression Is Cell Cycle Dependent in Trabecular Meshwork Cells |
| title_short | NFATc1 Regulation of Dexamethasone-Induced <i>TGFB2</i> Expression Is Cell Cycle Dependent in Trabecular Meshwork Cells |
| title_sort | nfatc1 regulation of dexamethasone induced i tgfb2 i expression is cell cycle dependent in trabecular meshwork cells |
| topic | trabecular meshwork TGFβ2 NFATc1 dexamethasone cell cycle quiescence |
| url | https://www.mdpi.com/2073-4409/12/3/504 |
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