Knock down of caveolin-1 affects morphological and functional hallmarks of human endothelial cells
Luca Madaro1, Fabrizio Antonangeli, Annarita Favia
1Department of Anatomy, Histology, Forensic Medicine and Orthopedics, Section of Histology and Medical Embryology, Sapienza University of Rome, Rome, Italy.
Abstract:
Caveolin-1 (CAV1) is the principal structural component of caveolae which functions as scaffolding protein for the integration of a variety of signaling pathways. In this study, we investigated the involvement of CAV1 in endothelial cell (EC) functions and show that siRNA-induced CAV1 silencing in the human EC line EA.hy926 induces distinctive morphological changes, such as a marked increase in cell size and formation of stress fibers. Design-based stereology was employed in this work to make unbiased quantification of morphometric properties such as volume, length, and surface of CAV1 silenced versus control cells. In addition, we showed that downregulation of CAV1 affects cell cycle progression at G1/S phase transition most likely by perturbation of AKT signaling. With the aim to assess the contribution of CAV1 to typical biological processes of EC, we report here that CAV1 targeting affects cell migration and matrix metalloproteinases (MMPs) activity, and reduces angiogenesis in response to VEGF, in vitro. Taken together our data suggest that the proper expression of CAV1 is important not only for maintaining the appropriate morphology and size of ECs but it might represent a prospective molecular target for studying key biological mechanisms such as senescence and tumorigenesis.
Insights
Caveolin-1 (CAV1) is crucial for endothelial cell (EC) morphology and function. Its silencing alters cell size, migration, and angiogenesis, suggesting CAV1 as a target for studying senescence and tumorigenesis.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Caveolin-1 (CAV1) is a key structural protein of caveolae.
- CAV1 acts as a scaffolding protein, integrating various signaling pathways.
Purpose of the Study:
- To investigate the role of CAV1 in endothelial cell (EC) functions.
- To analyze the impact of CAV1 silencing on EC morphology, cell cycle, migration, and angiogenesis.
Main Methods:
- Utilized siRNA to silence CAV1 in human EC line EA.hy926.
- Employed design-based stereology for unbiased morphometric quantification.
- Assessed cell cycle progression, AKT signaling, cell migration, MMP activity, and VEGF-induced angiogenesis.
Main Results:
- CAV1 silencing induced significant morphological changes, including increased cell size and stress fiber formation.
- Downregulation of CAV1 perturbed cell cycle progression at G1/S phase, likely via AKT signaling.
- CAV1 targeting reduced EC migration, matrix metalloproteinases (MMPs) activity, and in vitro angiogenesis.
Conclusions:
- Proper CAV1 expression is essential for maintaining EC morphology and size.
- CAV1 plays a significant role in EC migration, MMP activity, and angiogenesis.
- CAV1 may represent a molecular target for studying senescence and tumorigenesis.
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