Cysteine-rich 61 mediates inflammation by the NF-κB/cyclooxygenase-2 pathway in RF/6A cells
PURPOSE: Cysteine-rich 61 (Cyr61) may enhance angiogenesis and inflammation in diabetic retinopathy (DR). Cyclooxygenase-2 (COX-2) is an immediate-early gene product of inflammation and it also plays an important role in developing DR. We aim to investigate the effects of Cyr61 on COX-2 expression i...
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| Format: | Article |
| Language: | English |
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Wolters Kluwer Medknow Publications
2025-04-01
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| Series: | Taiwan Journal of Ophthalmology |
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| Online Access: | https://journals.lww.com/10.4103/tjo.TJO-D-24-00086 |
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| author | Po-Ting Yeh Jian-Jang You Chang-Hao Yang |
| author_facet | Po-Ting Yeh Jian-Jang You Chang-Hao Yang |
| author_sort | Po-Ting Yeh |
| collection | DOAJ |
| description | PURPOSE:
Cysteine-rich 61 (Cyr61) may enhance angiogenesis and inflammation in diabetic retinopathy (DR). Cyclooxygenase-2 (COX-2) is an immediate-early gene product of inflammation and it also plays an important role in developing DR. We aim to investigate the effects of Cyr61 on COX-2 expression in chorioretinal vascular endothelial (RF/6A) cells and to study the possible signal transduction pathway and the transcriptional mechanisms.
MATERIALS AND METHODS:
The effects of Cyr61 on COX-2 expression were evaluated via determining the mRNA, protein, and prostaglandin (PG) E2 levels of RF/6A cells. To examine the pathway in this process, RF/6A cells were pretreated with integrin ανβ3-blocking antibodies, a FAK inhibitor (PF573228), a PI3K inhibitor (LY294002), and an Akt inhibitor (A6730), respectively. Electrophoretic mobility shift assays (EMSAs) and luciferase reporter assays were applied to assess if NF-κB was involved in this response.
RESULTS:
Cyr61 stimulated the expression of COX-2 at the mRNA, protein, and PGE2 levels in a dose-dependent and time-dependent manner. Both COX-2 and PGE2 levels were attenuated during the response to Cyr61 stimulation by pretreatment with integrin ανβ3-blocking antibodies, PF573228, LY294002, and A6730 respectively. EMSA revealed that all of the aforementioned inhibitors suppressed NF-κB activation. Luciferase reporter assays further indicated that the mutation of the NF-κB-binding element in the COX-2 gene promoter reduced its gene expression.
CONCLUSIONS:
Induction of COX-2 by Cyr61 is mediated through the activation of the integrin ανβ3, FAK, PI3K/Akt, and NF-κB pathways in RF/6A cells. |
| format | Article |
| id | doaj-art-a99c1ac7f04e4017a0e9f0e22d31ae63 |
| institution | DOAJ |
| issn | 2211-5056 2211-5072 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Wolters Kluwer Medknow Publications |
| record_format | Article |
| series | Taiwan Journal of Ophthalmology |
| spelling | doaj-art-a99c1ac7f04e4017a0e9f0e22d31ae632025-08-20T02:44:09ZengWolters Kluwer Medknow PublicationsTaiwan Journal of Ophthalmology2211-50562211-50722025-04-0115229730710.4103/tjo.TJO-D-24-00086Cysteine-rich 61 mediates inflammation by the NF-κB/cyclooxygenase-2 pathway in RF/6A cellsPo-Ting YehJian-Jang YouChang-Hao YangPURPOSE: Cysteine-rich 61 (Cyr61) may enhance angiogenesis and inflammation in diabetic retinopathy (DR). Cyclooxygenase-2 (COX-2) is an immediate-early gene product of inflammation and it also plays an important role in developing DR. We aim to investigate the effects of Cyr61 on COX-2 expression in chorioretinal vascular endothelial (RF/6A) cells and to study the possible signal transduction pathway and the transcriptional mechanisms. MATERIALS AND METHODS: The effects of Cyr61 on COX-2 expression were evaluated via determining the mRNA, protein, and prostaglandin (PG) E2 levels of RF/6A cells. To examine the pathway in this process, RF/6A cells were pretreated with integrin ανβ3-blocking antibodies, a FAK inhibitor (PF573228), a PI3K inhibitor (LY294002), and an Akt inhibitor (A6730), respectively. Electrophoretic mobility shift assays (EMSAs) and luciferase reporter assays were applied to assess if NF-κB was involved in this response. RESULTS: Cyr61 stimulated the expression of COX-2 at the mRNA, protein, and PGE2 levels in a dose-dependent and time-dependent manner. Both COX-2 and PGE2 levels were attenuated during the response to Cyr61 stimulation by pretreatment with integrin ανβ3-blocking antibodies, PF573228, LY294002, and A6730 respectively. EMSA revealed that all of the aforementioned inhibitors suppressed NF-κB activation. Luciferase reporter assays further indicated that the mutation of the NF-κB-binding element in the COX-2 gene promoter reduced its gene expression. CONCLUSIONS: Induction of COX-2 by Cyr61 is mediated through the activation of the integrin ανβ3, FAK, PI3K/Akt, and NF-κB pathways in RF/6A cells.https://journals.lww.com/10.4103/tjo.TJO-D-24-00086aktcyclooxygenase-2cysteine-rich 61integrinnf-κbsignal transduction |
| spellingShingle | Po-Ting Yeh Jian-Jang You Chang-Hao Yang Cysteine-rich 61 mediates inflammation by the NF-κB/cyclooxygenase-2 pathway in RF/6A cells Taiwan Journal of Ophthalmology akt cyclooxygenase-2 cysteine-rich 61 integrin nf-κb signal transduction |
| title | Cysteine-rich 61 mediates inflammation by the NF-κB/cyclooxygenase-2 pathway in RF/6A cells |
| title_full | Cysteine-rich 61 mediates inflammation by the NF-κB/cyclooxygenase-2 pathway in RF/6A cells |
| title_fullStr | Cysteine-rich 61 mediates inflammation by the NF-κB/cyclooxygenase-2 pathway in RF/6A cells |
| title_full_unstemmed | Cysteine-rich 61 mediates inflammation by the NF-κB/cyclooxygenase-2 pathway in RF/6A cells |
| title_short | Cysteine-rich 61 mediates inflammation by the NF-κB/cyclooxygenase-2 pathway in RF/6A cells |
| title_sort | cysteine rich 61 mediates inflammation by the nf κb cyclooxygenase 2 pathway in rf 6a cells |
| topic | akt cyclooxygenase-2 cysteine-rich 61 integrin nf-κb signal transduction |
| url | https://journals.lww.com/10.4103/tjo.TJO-D-24-00086 |
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