Centrosomal Protein CEP135 Regulates the Migration and Angiogenesis of Endothelial Cells in a Microtubule-Dependent Manner

Abstract

Background: Angiogenesis is the formation of blood vessels by sprouting from mature blood vessels and is regulated by multiple factors; however, the role of centrosome and centrosomal proteins (CEP) in angiogenesis needs further study. centrosomal protein 135 (CEP135) is an important centrosome-associated protein that can affect a variety of cellular processes, such as the cell cycle, but its effect on angiogenesis is still unknown.

Methods: Tube formation and in vivo angiogenesis assays were performed to confirm the effects of CEP135 on endothelial cell (EC) angiogenesis in vitro and in mice. Cell counting kit-8 (CCK-8), 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), flow cytometry (FCM) and immunoblot assays were performed to confirm the effects of CEP135 on the proliferation and cell cycle of endothelial cells. Wound healing, transwell, and fluorescence staining were performed to confirm its effects on EC cell migration, polarization, and spindle orientation, and a tubulin turbidity assay was performed to confirm its effects on microtubule stabilization.

Results: We conducted a series of experiments to explore the potential role of CEP135 in angiogenesis. CEP135 siRNA obviously inhibits angiogenesis in vivo compared with the control. CEP135 could mediate cell proliferation and the cell cycle by mediating spindle orientation. Moreover, we showed that CEP135 is involved in the regulation of angiogenesis by affecting the migration of endothelial cells using wound closure and transwell assays. We further demonstrated that CEP135 promotes endothelial polarization and microtubule stability, thus affecting cell migration.

Conclusions: CEP135 was involved in the polarization of centrosomes, which is important for the migration of human umbilical vein ECs (HUVECs). These findings indicated that CEP135 may promote the polarization of HUVECs and accelerate migration, which in turn promotes angiogenesis.

Keywords: CEP135; angiogenesis; endothelial cells; migration; polarization.