Prolidase Stimulates Proliferation and Migration through Activation of the PI3K/Akt/mTOR Signaling Pathway in Human Keratinocytes
Recent reports have indicated prolidase (PEPD) as a ligand of the epidermal growth factor receptor (EGFR). Since this receptor is involved in the promotion of cell proliferation, growth, and migration, we aimed to investigate whether prolidase may participate in wound healing in vitro. All experimen...
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| Format: | Article |
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MDPI AG
2020-12-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/21/23/9243 |
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| author | Magdalena Misiura Weronika Baszanowska Ilona Ościłowska Jerzy Pałka Wojciech Miltyk |
| author_facet | Magdalena Misiura Weronika Baszanowska Ilona Ościłowska Jerzy Pałka Wojciech Miltyk |
| author_sort | Magdalena Misiura |
| collection | DOAJ |
| description | Recent reports have indicated prolidase (PEPD) as a ligand of the epidermal growth factor receptor (EGFR). Since this receptor is involved in the promotion of cell proliferation, growth, and migration, we aimed to investigate whether prolidase may participate in wound healing in vitro. All experiments were performed in prolidase-treated human keratinocytes assessing cell vitality, proliferation, and migration. The expression of downstream signaling proteins induced by EGFR, insulin-like growth factor 1 (IGF-1), transforming growth factor β<sub>1</sub> (TGF-β<sub>1</sub>), and β<sub>1</sub>-integrin receptors were evaluated by Western immunoblotting and immunocytochemical staining. To determine collagen biosynthesis and prolidase activity radiometric and colorimetric methods were used, respectively. Proline content was determined by applying the liquid chromatography coupled with mass spectrometry. We found that prolidase promoted the proliferation and migration of keratinocytes through stimulation of EGFR-downstream signaling pathways in which the PI3K/Akt/mTOR axis was involved. Moreover, PEPD upregulated the expression of β<sub>1</sub>-integrin and IGF-1 receptors and their downstream proteins. Proline concentration and collagen biosynthesis were increased in HaCaT cells under prolidase treatment. Since extracellular prolidase as a ligand of EGFR induced cell growth, migration, and collagen biosynthesis in keratinocytes, it may represent a potential therapeutic approach for the treatment of skin wounds. |
| first_indexed | 2024-03-10T14:20:27Z |
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| id | doaj.art-5e8198b4bae846d6a3fa1e277d11a261 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T14:20:27Z |
| publishDate | 2020-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-5e8198b4bae846d6a3fa1e277d11a2612023-11-20T23:26:29ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-12-012123924310.3390/ijms21239243Prolidase Stimulates Proliferation and Migration through Activation of the PI3K/Akt/mTOR Signaling Pathway in Human KeratinocytesMagdalena Misiura0Weronika Baszanowska1Ilona Ościłowska2Jerzy Pałka3Wojciech Miltyk4Department of Analysis and Bioanalysis of Medicines, Medical University of Bialystok, Jana Kilińskiego 1, 15-089 Bialystok, PolandDepartment of Medicinal Chemistry, Medical University of Bialystok, Jana Kilińskiego 1, 15-089 Bialystok, PolandDepartment of Medicinal Chemistry, Medical University of Bialystok, Jana Kilińskiego 1, 15-089 Bialystok, PolandDepartment of Medicinal Chemistry, Medical University of Bialystok, Jana Kilińskiego 1, 15-089 Bialystok, PolandDepartment of Analysis and Bioanalysis of Medicines, Medical University of Bialystok, Jana Kilińskiego 1, 15-089 Bialystok, PolandRecent reports have indicated prolidase (PEPD) as a ligand of the epidermal growth factor receptor (EGFR). Since this receptor is involved in the promotion of cell proliferation, growth, and migration, we aimed to investigate whether prolidase may participate in wound healing in vitro. All experiments were performed in prolidase-treated human keratinocytes assessing cell vitality, proliferation, and migration. The expression of downstream signaling proteins induced by EGFR, insulin-like growth factor 1 (IGF-1), transforming growth factor β<sub>1</sub> (TGF-β<sub>1</sub>), and β<sub>1</sub>-integrin receptors were evaluated by Western immunoblotting and immunocytochemical staining. To determine collagen biosynthesis and prolidase activity radiometric and colorimetric methods were used, respectively. Proline content was determined by applying the liquid chromatography coupled with mass spectrometry. We found that prolidase promoted the proliferation and migration of keratinocytes through stimulation of EGFR-downstream signaling pathways in which the PI3K/Akt/mTOR axis was involved. Moreover, PEPD upregulated the expression of β<sub>1</sub>-integrin and IGF-1 receptors and their downstream proteins. Proline concentration and collagen biosynthesis were increased in HaCaT cells under prolidase treatment. Since extracellular prolidase as a ligand of EGFR induced cell growth, migration, and collagen biosynthesis in keratinocytes, it may represent a potential therapeutic approach for the treatment of skin wounds.https://www.mdpi.com/1422-0067/21/23/9243prolidasePEPDEGFRkeratinocyteswound healing |
| spellingShingle | Magdalena Misiura Weronika Baszanowska Ilona Ościłowska Jerzy Pałka Wojciech Miltyk Prolidase Stimulates Proliferation and Migration through Activation of the PI3K/Akt/mTOR Signaling Pathway in Human Keratinocytes International Journal of Molecular Sciences prolidase PEPD EGFR keratinocytes wound healing |
| title | Prolidase Stimulates Proliferation and Migration through Activation of the PI3K/Akt/mTOR Signaling Pathway in Human Keratinocytes |
| title_full | Prolidase Stimulates Proliferation and Migration through Activation of the PI3K/Akt/mTOR Signaling Pathway in Human Keratinocytes |
| title_fullStr | Prolidase Stimulates Proliferation and Migration through Activation of the PI3K/Akt/mTOR Signaling Pathway in Human Keratinocytes |
| title_full_unstemmed | Prolidase Stimulates Proliferation and Migration through Activation of the PI3K/Akt/mTOR Signaling Pathway in Human Keratinocytes |
| title_short | Prolidase Stimulates Proliferation and Migration through Activation of the PI3K/Akt/mTOR Signaling Pathway in Human Keratinocytes |
| title_sort | prolidase stimulates proliferation and migration through activation of the pi3k akt mtor signaling pathway in human keratinocytes |
| topic | prolidase PEPD EGFR keratinocytes wound healing |
| url | https://www.mdpi.com/1422-0067/21/23/9243 |
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