論文 - 詳細
| RRC ID | 54033 |
|---|---|
| 著者 | Gillies PJ, Richardson NA, Walshe J, Stephenson SA, Dawson RA, Harkin DG. |
| タイトル | Demonstration of P-selectin expression and potential function in human corneal epithelial cells. |
| ジャーナル | Exp Eye Res |
| Abstract |
In response to an unexpected observation of apparent localisation by immunocytochemistry, we have investigated the potential expression and function of P-selectin (CD62P) in human corneal epithelial cells. The SV40 immortalised cell line, HCE-T (validated by STR profiling), along with multiple donor corneal-limbal tissue samples, were examined for P-selectin expression using a combination of immunocytochemistry, Western blotting, RT-PCR and immunohistochemistry. Potential expression of the major ligand for P-selectin (P-selectin glycoprotein ligand-1; PSGL-1; CD162) was also examined by immunocytochemistry and RT-PCR. A selective inhibitor of P-selectin-PSGL-1 binding (KF38789) was subsequently tested for effects on HCE-T cells using a cell culture gap-closure assay. HCE-T cells as well as primary epithelial cultures derived from donor corneal-limbal tissue, displayed positive immunostaining for P-selectin. Staining was particularly evident at cell-cell boundaries and at the outer edge of expanding epithelial islands. P-selectin expression was confirmed by Western blotting and RT-PCR (validated by product sequencing), as well as by immunohistochemistry performed on serial sections of corneal-limbal tissue stained for P-selectin, keratin 3 and p63. PSGL-1 was detected by RT-PCR and immunocytochemistry in both corneal epithelial cells as well as human limbal fibroblasts (HLF). KF38789 (5 μM) significantly reduced closure of a 500-μm gap between confluent sheets of HCE-T cells over an 8-hr period (by ∼40%, p < 0.01; paired two-tailed T test), but had no effect on culture gap-closure by either HLF or murine 3T3 fibroblasts. These results provide evidence of P-selectin expression in human corneal epithelial cells and suggest a potential role for this glycoprotein in facilitating the net movement of confluent sheets of human corneal epithelial cells. |
| 巻・号 | 176 |
| ページ | 196-206 |
| 公開日 | 2018-11-1 |
| DOI | 10.1016/j.exer.2018.07.018 |
| PII | S0014-4835(18)30174-X |
| PMID | 30025918 |
| MeSH | Biomarkers / metabolism Blotting, Western Cell Separation / methods Cells, Cultured Epithelium, Corneal / metabolism* Fibroblasts / metabolism Gene Expression / physiology Humans Immunohistochemistry Limbus Corneae / cytology Membrane Glycoproteins / metabolism P-Selectin / genetics* P-Selectin / metabolism* Real-Time Polymerase Chain Reaction |
| IF | 3.011 |
| 引用数 | 0 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 8 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | HCE-T(RCB2280) |