Rabbit Anti-Human PAK1 Polyclonal antibody

Background: MicroRNA-146a (miR-146a) is reportedly implicated in the pathogenesis of cancer cells. Its role in the human coronary artery endothelial cells (HCAECs) is however unclear. In this study, the effects of miR-146a on the proliferation and angiogenesis of HCAECs were investigated. Methods: MiR-146a was up- or down-regulated by transfection of mimic or inhibitor. BrdU assay was used to evaluate the proliferation of HCAECs. Wound healing assay and Transwell penetrating experiment were performed to investigate the migration and penetrating ability. We investigated whether miR-146a influences the secretion of vascular endothelial growth factor (VEGF) and angiogenesis of HCAECs. Regulatory mechanism of miR-146a was investigated meanwhile. Results: MiR-146a mimic promote the proliferation, migration, penetrating, angiogenesis ability of HCAECs. Overexpression of miR-146a decreased the expression of neurofibromin 2, but increased the expression of PAK1 and VEGF followed with the enhancement of angiogenesis. The up-regulation effect of VEGF by miR-146a can be blocked by overexpression of NF2. Conclusions: MiR-146a can increase the proliferation, migration and penetrating ability of HCAECs, and promote angiogenesis by up-regulating expression of VEGF via NF2/PAKJ pathway. Upregulation of miR-146a may be a new therapeutic target to improve the blood supply for acute myocardial infarction.

Atypical RhoV GTPase (Chp/Wrch-2) is a member of the human Rho GTPase family, which belongs to the superfamily of Ras-related small GTPases. The biological functions of RhoV, regulation of its activity, and mechanisms of its action remain largely unexplored. Rho GTPases regulate a wide range of cellular processes by interacting with protein targets called effectors. Several putative RhoV effectors have been identified, including protein kinases of the Pak (p21-activated kinase) family: Pak1, Pak2, Pak4, and Pak6. RhoV GTPase activates Pak1 protein kinase and simultaneously induces its ubiquitin-dependent degradation. Pak1 regulates E-cadherin localization at adherens junctions downstream of RhoV during gastrulation in fish. The effector domain of RhoV mediates its binding to the CRIB (Cdc42/Rac1 interactive binding) motif in the N-terminal p21-binding domain (PBD) of Pak6 protein kinase. The role of the RhoV effector domain in mediating interaction with Pak1 has not been studied. This study has identified mutations in the effector domain of RhoV GTPase (Y60K, T63A, L65A, and D66A) that impair its interaction with Pak1 in the GST-PAK-PBD pull-down assay and coimmunoprecipitation. Our results suggest that the effector domain of RhoV mediates its binding to Pak1, complementing the current view of the molecular basics of RhoV binding to effectors of the Pak family. These data lay the basis for further studies on the role of Pak1 in RhoV-activated signaling pathways and cellular processes.