Caveolin-1 Regulates P2Y<sub>2</sub> Receptor Signaling during Mechanical Injury in Human 1321N1 Astrocytoma
Caveolae-associated protein caveolin-1 (Cav-1) plays key roles in cellular processes such as mechanosensing, receptor coupling to signaling pathways, cell growth, apoptosis, and cancer. In 1321N1 astrocytoma cells Cav-1 interacts with the P2Y<sub>2</sub> receptor (P2Y<sub>2</sub...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2019-10-01
|
| Series: | Biomolecules |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2218-273X/9/10/622 |
| _version_ | 1818541102230142976 |
|---|---|
| author | Magdiel Martínez Namyr A. Martínez Jorge D. Miranda Héctor M. Maldonado Walter I. Silva Ortiz |
| author_facet | Magdiel Martínez Namyr A. Martínez Jorge D. Miranda Héctor M. Maldonado Walter I. Silva Ortiz |
| author_sort | Magdiel Martínez |
| collection | DOAJ |
| description | Caveolae-associated protein caveolin-1 (Cav-1) plays key roles in cellular processes such as mechanosensing, receptor coupling to signaling pathways, cell growth, apoptosis, and cancer. In 1321N1 astrocytoma cells Cav-1 interacts with the P2Y<sub>2</sub> receptor (P2Y<sub>2</sub>R) to modulate its downstream signaling. P2Y<sub>2</sub>R and its signaling machinery also mediate pro-survival actions after mechanical injury. This study determines if Cav-1 knockdown (KD) affects P2Y<sub>2</sub>R signaling and its pro-survival actions in the 1321N1 astrocytoma cells mechanical injury model system. KD of Cav-1 decreased its expression in 1321N1 cells devoid of or expressing hHAP2Y<sub>2</sub>R by ~88% and ~85%, respectively. Cav-1 KD had no significant impact on P2Y<sub>2</sub>R expression. Post-injury densitometric analysis of pERK1/2 and Akt activities in Cav-1-positive 1321N1 cells (devoid of or expressing a hHAP2Y<sub>2</sub>R) revealed a P2Y2R-dependent temporal increase in both kinases. These temporal increases in pERK1/2 and pAkt were significantly decreased in Cav-1 KD 1321N1 (devoid of or expressing a hHAP2Y<sub>2</sub>R). Cav-1 KD led to an ~2.0-fold and ~2.4-fold decrease in the magnitude of the hHAP2Y<sub>2</sub>R-mediated pERK1/2 and pAkt kinases’ activity, respectively. These early-onset hHAP2Y<sub>2</sub>R-mediated signaling responses in Cav-1-expressing and Cav-1 KD 1321N1 correlated with changes in cell viability (via a resazurin-based method) and apoptosis (via caspase-9 expression). In Cav-1-positive 1321N1 cells, expression of hHAP2Y<sub>2</sub>R led to a significant increase in cell viability and decreased apoptotic (caspase-9) activity after mechanical injury. In contrast, hHAP2Y<sub>2</sub>R-elicited changes in viability and apoptotic (caspase-9) activity were decreased after mechanical injury in Cav-1 KD 1321N1 cells expressing hHAP2Y<sub>2</sub>R. These findings support the importance of Cav-1 in modulating P2Y<sub>2</sub>R signaling during mechanical injury and its protective actions in a human astrocytoma cell line, whilst shedding light on potential new venues for brain injury or trauma interventions. |
| first_indexed | 2024-12-11T22:04:09Z |
| format | Article |
| id | doaj.art-b7e08fab00984ba29ae40335d0364650 |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-12-11T22:04:09Z |
| publishDate | 2019-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-b7e08fab00984ba29ae40335d03646502022-12-22T00:49:00ZengMDPI AGBiomolecules2218-273X2019-10-0191062210.3390/biom9100622biom9100622Caveolin-1 Regulates P2Y<sub>2</sub> Receptor Signaling during Mechanical Injury in Human 1321N1 AstrocytomaMagdiel Martínez0Namyr A. Martínez1Jorge D. Miranda2Héctor M. Maldonado3Walter I. Silva Ortiz4Physiology Department, University of Puerto Rico, School of Medicine, San Juan 00936-5067, Puerto RicoPhysiology Department, University of Puerto Rico, School of Medicine, San Juan 00936-5067, Puerto RicoPhysiology Department, University of Puerto Rico, School of Medicine, San Juan 00936-5067, Puerto RicoPharmacology Department, Universidad Central del Caribe, Bayamón 00960-6032, Puerto RicoPhysiology Department, University of Puerto Rico, School of Medicine, San Juan 00936-5067, Puerto RicoCaveolae-associated protein caveolin-1 (Cav-1) plays key roles in cellular processes such as mechanosensing, receptor coupling to signaling pathways, cell growth, apoptosis, and cancer. In 1321N1 astrocytoma cells Cav-1 interacts with the P2Y<sub>2</sub> receptor (P2Y<sub>2</sub>R) to modulate its downstream signaling. P2Y<sub>2</sub>R and its signaling machinery also mediate pro-survival actions after mechanical injury. This study determines if Cav-1 knockdown (KD) affects P2Y<sub>2</sub>R signaling and its pro-survival actions in the 1321N1 astrocytoma cells mechanical injury model system. KD of Cav-1 decreased its expression in 1321N1 cells devoid of or expressing hHAP2Y<sub>2</sub>R by ~88% and ~85%, respectively. Cav-1 KD had no significant impact on P2Y<sub>2</sub>R expression. Post-injury densitometric analysis of pERK1/2 and Akt activities in Cav-1-positive 1321N1 cells (devoid of or expressing a hHAP2Y<sub>2</sub>R) revealed a P2Y2R-dependent temporal increase in both kinases. These temporal increases in pERK1/2 and pAkt were significantly decreased in Cav-1 KD 1321N1 (devoid of or expressing a hHAP2Y<sub>2</sub>R). Cav-1 KD led to an ~2.0-fold and ~2.4-fold decrease in the magnitude of the hHAP2Y<sub>2</sub>R-mediated pERK1/2 and pAkt kinases’ activity, respectively. These early-onset hHAP2Y<sub>2</sub>R-mediated signaling responses in Cav-1-expressing and Cav-1 KD 1321N1 correlated with changes in cell viability (via a resazurin-based method) and apoptosis (via caspase-9 expression). In Cav-1-positive 1321N1 cells, expression of hHAP2Y<sub>2</sub>R led to a significant increase in cell viability and decreased apoptotic (caspase-9) activity after mechanical injury. In contrast, hHAP2Y<sub>2</sub>R-elicited changes in viability and apoptotic (caspase-9) activity were decreased after mechanical injury in Cav-1 KD 1321N1 cells expressing hHAP2Y<sub>2</sub>R. These findings support the importance of Cav-1 in modulating P2Y<sub>2</sub>R signaling during mechanical injury and its protective actions in a human astrocytoma cell line, whilst shedding light on potential new venues for brain injury or trauma interventions.https://www.mdpi.com/2218-273X/9/10/622caveolin-1p2y2signalingcaveolaebrain injuryastrocytoma |
| spellingShingle | Magdiel Martínez Namyr A. Martínez Jorge D. Miranda Héctor M. Maldonado Walter I. Silva Ortiz Caveolin-1 Regulates P2Y<sub>2</sub> Receptor Signaling during Mechanical Injury in Human 1321N1 Astrocytoma Biomolecules caveolin-1 p2y2 signaling caveolae brain injury astrocytoma |
| title | Caveolin-1 Regulates P2Y<sub>2</sub> Receptor Signaling during Mechanical Injury in Human 1321N1 Astrocytoma |
| title_full | Caveolin-1 Regulates P2Y<sub>2</sub> Receptor Signaling during Mechanical Injury in Human 1321N1 Astrocytoma |
| title_fullStr | Caveolin-1 Regulates P2Y<sub>2</sub> Receptor Signaling during Mechanical Injury in Human 1321N1 Astrocytoma |
| title_full_unstemmed | Caveolin-1 Regulates P2Y<sub>2</sub> Receptor Signaling during Mechanical Injury in Human 1321N1 Astrocytoma |
| title_short | Caveolin-1 Regulates P2Y<sub>2</sub> Receptor Signaling during Mechanical Injury in Human 1321N1 Astrocytoma |
| title_sort | caveolin 1 regulates p2y sub 2 sub receptor signaling during mechanical injury in human 1321n1 astrocytoma |
| topic | caveolin-1 p2y2 signaling caveolae brain injury astrocytoma |
| url | https://www.mdpi.com/2218-273X/9/10/622 |
| work_keys_str_mv | AT magdielmartinez caveolin1regulatesp2ysub2subreceptorsignalingduringmechanicalinjuryinhuman1321n1astrocytoma AT namyramartinez caveolin1regulatesp2ysub2subreceptorsignalingduringmechanicalinjuryinhuman1321n1astrocytoma AT jorgedmiranda caveolin1regulatesp2ysub2subreceptorsignalingduringmechanicalinjuryinhuman1321n1astrocytoma AT hectormmaldonado caveolin1regulatesp2ysub2subreceptorsignalingduringmechanicalinjuryinhuman1321n1astrocytoma AT walterisilvaortiz caveolin1regulatesp2ysub2subreceptorsignalingduringmechanicalinjuryinhuman1321n1astrocytoma |