MiR-199a-3p Induces Mesenchymal to Epithelial Transition of Keratinocytes by Targeting RAP2B
Cutaneous squamous cell carcinoma (CSCC) is an epidermal skin cancer that evolves from normal epidermis along several pre-malignant stages. Previously we found specific miRNAs alterations in each step along these stages. miR-199a-3p expression decreases at the transition to later stages. A crucial s...
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MDPI AG
2022-12-01
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| Series: | International Journal of Molecular Sciences |
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| author | Moamen Masalha Tal Meningher Adi Mizrahi Aviv Barzilai Hilla Tabibian-Keissar Devorah Gur-Wahnon Iddo Z. Ben-Dov Joshua Kapenhas Jasmine Jacob-Hirsch Raya Leibowitz Yechezkel Sidi Dror Avni |
| author_facet | Moamen Masalha Tal Meningher Adi Mizrahi Aviv Barzilai Hilla Tabibian-Keissar Devorah Gur-Wahnon Iddo Z. Ben-Dov Joshua Kapenhas Jasmine Jacob-Hirsch Raya Leibowitz Yechezkel Sidi Dror Avni |
| author_sort | Moamen Masalha |
| collection | DOAJ |
| description | Cutaneous squamous cell carcinoma (CSCC) is an epidermal skin cancer that evolves from normal epidermis along several pre-malignant stages. Previously we found specific miRNAs alterations in each step along these stages. miR-199a-3p expression decreases at the transition to later stages. A crucial step for epithelial carcinoma cells to acquire invasive capacity is the disruption of cell–cell contacts and the gain of mesenchymal motile phenotype, a process known as epithelial-to-mesenchymal transition (EMT). This study aims to study the role of decreased expression of miR-199a-3p in keratinocytes’ EMT towards carcinogenesis. First, we measured miR-199a-3p in different stages of epidermal carcinogenesis. Then, we applied Photoactivatable Ribonucleoside-Enhanced Crosslinking and Immunoprecipitation (PAR-CLIP) assay to search for possible biochemical targets of miR-199a-3p and verified that Ras-associated protein B2 (RAP2B) is a bona-fide target of miR-199a-3p. Next, we analyzed RAP2B expression, in CSCC biopsies. Last, we evaluated possible mechanisms leading to decreased miR-199a-3p expression. miR-199a-3p induces a mesenchymal to epithelial transition (MET) in CSSC cells. Many of the under-expressed genes in CSCC overexpressing miR-199a-3p, are possible targets of miR-199a-3p and play roles in EMT. RAP2B is a biochemical target of miR-199a-3p. Overexpression of miR-199a-3p in CSCC results in decreased phosphorylated focal adhesion kinase (FAK). In addition, inhibiting FAK phosphorylation inhibits EMT marker genes’ expression. In addition, we proved that DNA methylation is part of the mechanism by which miR-199a-3p expression is inhibited. However, it is not by the methylation of miR-199a putative promoter. These findings suggest that miR-199a-3p inhibits the EMT process by targeting RAP2B. Inhibitors of RAP2B or FAK may be effective therapeutic agents for CSCC. |
| first_indexed | 2024-03-09T17:44:09Z |
| format | Article |
| id | doaj.art-067e2d2069e44876adaf505edbde5d0e |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T17:44:09Z |
| publishDate | 2022-12-01 |
| publisher | MDPI AG |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-067e2d2069e44876adaf505edbde5d0e2023-11-24T11:18:28ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-12-0123231540110.3390/ijms232315401MiR-199a-3p Induces Mesenchymal to Epithelial Transition of Keratinocytes by Targeting RAP2BMoamen Masalha0Tal Meningher1Adi Mizrahi2Aviv Barzilai3Hilla Tabibian-Keissar4Devorah Gur-Wahnon5Iddo Z. Ben-Dov6Joshua Kapenhas7Jasmine Jacob-Hirsch8Raya Leibowitz9Yechezkel Sidi10Dror Avni11Laboratory of Molecular Cell Biology, Center for Cancer Research, Department of Medicine C, Sheba Medical Center, Tel Hashomer 52621, IsraelLaboratory of Molecular Cell Biology, Center for Cancer Research, Department of Medicine C, Sheba Medical Center, Tel Hashomer 52621, IsraelLaboratory of Molecular Cell Biology, Center for Cancer Research, Department of Medicine C, Sheba Medical Center, Tel Hashomer 52621, IsraelDepartment of Dermatology, Institute of Pathology Sheba Medical Center, Tel Hashomer 52621, IsraelDepartment of Pathology, Sheba Medical Center, Tel Hashomer 52621, IsraelLaboratory of Medical Transcriptomics, Nephrology and Hypertension Services, Hadassah-Hebrew University Medical Center, Jerusalem 91120, IsraelLaboratory of Medical Transcriptomics, Nephrology and Hypertension Services, Hadassah-Hebrew University Medical Center, Jerusalem 91120, IsraelLaboratory of Molecular Cell Biology, Center for Cancer Research, Department of Medicine C, Sheba Medical Center, Tel Hashomer 52621, IsraelCenter for Cancer Research Sheba Medical Center, Tel Hashomer 52621, IsraelFaculty of Medicine, Sackler School of Medicine, Tel Aviv University, Tel Aviv 69978, IsraelLaboratory of Molecular Cell Biology, Center for Cancer Research, Department of Medicine C, Sheba Medical Center, Tel Hashomer 52621, IsraelLaboratory of Molecular Cell Biology, Center for Cancer Research, Department of Medicine C, Sheba Medical Center, Tel Hashomer 52621, IsraelCutaneous squamous cell carcinoma (CSCC) is an epidermal skin cancer that evolves from normal epidermis along several pre-malignant stages. Previously we found specific miRNAs alterations in each step along these stages. miR-199a-3p expression decreases at the transition to later stages. A crucial step for epithelial carcinoma cells to acquire invasive capacity is the disruption of cell–cell contacts and the gain of mesenchymal motile phenotype, a process known as epithelial-to-mesenchymal transition (EMT). This study aims to study the role of decreased expression of miR-199a-3p in keratinocytes’ EMT towards carcinogenesis. First, we measured miR-199a-3p in different stages of epidermal carcinogenesis. Then, we applied Photoactivatable Ribonucleoside-Enhanced Crosslinking and Immunoprecipitation (PAR-CLIP) assay to search for possible biochemical targets of miR-199a-3p and verified that Ras-associated protein B2 (RAP2B) is a bona-fide target of miR-199a-3p. Next, we analyzed RAP2B expression, in CSCC biopsies. Last, we evaluated possible mechanisms leading to decreased miR-199a-3p expression. miR-199a-3p induces a mesenchymal to epithelial transition (MET) in CSSC cells. Many of the under-expressed genes in CSCC overexpressing miR-199a-3p, are possible targets of miR-199a-3p and play roles in EMT. RAP2B is a biochemical target of miR-199a-3p. Overexpression of miR-199a-3p in CSCC results in decreased phosphorylated focal adhesion kinase (FAK). In addition, inhibiting FAK phosphorylation inhibits EMT marker genes’ expression. In addition, we proved that DNA methylation is part of the mechanism by which miR-199a-3p expression is inhibited. However, it is not by the methylation of miR-199a putative promoter. These findings suggest that miR-199a-3p inhibits the EMT process by targeting RAP2B. Inhibitors of RAP2B or FAK may be effective therapeutic agents for CSCC.https://www.mdpi.com/1422-0067/23/23/15401cutaneous squamous cell carcinoma (CSCC)miR-199a-3pepithelial-mesenchymal transition (EMT)RAP2B |
| spellingShingle | Moamen Masalha Tal Meningher Adi Mizrahi Aviv Barzilai Hilla Tabibian-Keissar Devorah Gur-Wahnon Iddo Z. Ben-Dov Joshua Kapenhas Jasmine Jacob-Hirsch Raya Leibowitz Yechezkel Sidi Dror Avni MiR-199a-3p Induces Mesenchymal to Epithelial Transition of Keratinocytes by Targeting RAP2B International Journal of Molecular Sciences cutaneous squamous cell carcinoma (CSCC) miR-199a-3p epithelial-mesenchymal transition (EMT) RAP2B |
| title | MiR-199a-3p Induces Mesenchymal to Epithelial Transition of Keratinocytes by Targeting RAP2B |
| title_full | MiR-199a-3p Induces Mesenchymal to Epithelial Transition of Keratinocytes by Targeting RAP2B |
| title_fullStr | MiR-199a-3p Induces Mesenchymal to Epithelial Transition of Keratinocytes by Targeting RAP2B |
| title_full_unstemmed | MiR-199a-3p Induces Mesenchymal to Epithelial Transition of Keratinocytes by Targeting RAP2B |
| title_short | MiR-199a-3p Induces Mesenchymal to Epithelial Transition of Keratinocytes by Targeting RAP2B |
| title_sort | mir 199a 3p induces mesenchymal to epithelial transition of keratinocytes by targeting rap2b |
| topic | cutaneous squamous cell carcinoma (CSCC) miR-199a-3p epithelial-mesenchymal transition (EMT) RAP2B |
| url | https://www.mdpi.com/1422-0067/23/23/15401 |
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