The effect of RhoA on human umbilical vein endothelial cell migration and angiogenesis in vitro

Abstract

Abstract. The mechanisms that control the morphologic organization of endothelial cells (ECs) into new blood vessels are not well understood. Recent studies revealed that the small G proteins of the Rho family are key regulators of cell migration, involving reorganization of the actin cytoskeleton, cell migration and the regulation of gene transcription. We hypothesized that RhoA GTPase, a member of the Rho family, may play an important role in EC organization during angiogenesis, the process of new vessel formation in preexisting tissues. To test this hypothesis, we investigated the effects of RhoA on human umbilical vein endothelial (HUVE) cell migration and angiogenesis in vitro, by stably transfecting HUVE cells with sense RhoA expression plasmid through the Lipofect-2000 system. Wound assay in vitro and 3-dimensional cell culture were used to detect the migration and angiogenesis capacity of HUVE cells. The morphological changes of transfected cells were revealed under confocal and phase contrast microscopy. Our results demonstrated that the increased expression of RhoA in HUVE cells significantly enhanced the morphogenetic changes and cytoskeletal reorganization of the transfected cells, and also enhanced cell migration and angiogenic capacity in vitro, suggesting that RhoA plays an important role in the process of HUVE cell migration and angiogenesis in vitro.

Inhibition of the PI3K–Akt and mTORC1 signaling pathways promotes the elongation of vascular endothelial cells

“…The Rho-ROCK pathway regulates the actin cytoskeleton of endothelial cells, thereby controlling angiogenesis (Chen et al, 2014;van Nieuw Amerongen et al, 2003;Zhao et al, 2006). In line with the proposed role of the Rho-ROCK pathway, inhibition of ROCK activity by the specific inhibitor, thiazovivin, blocked the elongation of endothelial cells.…”

“…VEGF mediates tumor angiogenesis by activating the phosphatidylinositol 3-kinase (PI3K) to Akt signaling pathway (Cross and Claesson-Welsh, 2001;Gerber et al, 1998). PI3K-Akt signaling also indirectly upregulates tumor angiogenesis through activation of the mammalian target of rapamycin complex 1 (mTORC1) (Jiang et al, 2002;Karar and Maity, 2011;Laughner et al, 2001;Okumura et al, 2012;Zhao et al, 2006). mTORC1 increases the level of hypoxia inducible factor 1α (HIF-1α) in tumor cells, thereby promoting VEGF secretion and the formation of leaky tumor vessels.…”

Fasudil inhibits lung carcinoma-conditioned endothelial cell viability and migration

“…Thus, the function of RhoA is consistent with a possible role in vascular morphogenesis. ROCK may be a key player in ECs during angiogenesis (26). ROCK mediated angiogenesis plays an important role in tumour progression and metastasis, and is an attractive target for anticancer strategies.…”

Glabridin Inhibits Migration, Invasion, and Angiogenesis of Human Non–Small Cell Lung Cancer A549 Cells by Inhibiting the FAK/Rho Signaling Pathway

“…27,28 Upregulation of RhoA in HUVEC significantly enhanced morphogenetic changes and cytoskeletal reorganization, resulting in enhanced cell migration and angiogenic capacity. 29 Aberrant regulation of Rho proteins is associated with metastasis by promoting cancer angiogenesis and tumor cell motility. 29 One of the major downstream effectors of RhoA with regard to the cell migration is Rho kinase (ROCK), which can phosphorylate myosin light chain (MLC) at Thr 18/Ser 19 and subsequently trigger myosin contraction, leading to cell migration.…”

“…29 Aberrant regulation of Rho proteins is associated with metastasis by promoting cancer angiogenesis and tumor cell motility. 29 One of the major downstream effectors of RhoA with regard to the cell migration is Rho kinase (ROCK), which can phosphorylate myosin light chain (MLC) at Thr 18/Ser 19 and subsequently trigger myosin contraction, leading to cell migration. 30 Our results show that glabridin treatment decreases the activation of Akt, followed by a decrease of Rho activity.…”