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To investigate the effect of WWC3 on vascular smooth muscle cells (VSMCs) after injury and its possible mechanism.
We used platelet-derived growth factor BB (PDGF-BB) as cell injury, rats with balloon injury to make model of rat carotid artery intima injury. Detect the expression of WWC3 in A10 cells (VSMCs) and arteries after injury, and the influence on the proliferation and migration of VSMCs by MTT, Transwell, Western blot, immunohistochemical, and other methods. By regulating WWC3 expression, with luciferase activity assay, immunoprecipitation and other methods, to investigate the molecular mechanism of WWC3 on Hippo signaling pathway.
After injury, the expression of WWC3 decreased in A10 cells and rat carotid artery, Hippo signaling pathway activity down-regulated significantly, the expression of YAP (main effector of Hippo signaling pathway), and its main negative downstream target genes such as CTGF increased, enhancing the proliferation WWC3. Overexpressing WWC3, meanwhile stimulating with PDGF-BB, the proliferation and migration of VSMCs were significantly weakened, compared with stimulating with PDGF-BB alone. It is confirmed that WWC3 could interact with LATS1 by co-immunoprecipitation, induce phosphorylation of LATS1, inhibit the nuclear import of YAP with phosphorylation of YAP, thereby upregulate Hippo pathway activity. However, transfecting WWC3-ΔWW, this effect disappeared.
WWC3 expression is down-regulated in VSMCs and the neointimal hyperplasia after the injury (with PDGF-BB or balloon injury), inhibiting the activity of Hippo signaling pathway, enhancing the ability of proliferation and migration of VSMCs. WWC3 can interact with LATS1, to promote the phosphorylation of YAP and reduce its nuclear transference, upregulating Hippo signaling pathway activity, and inhibiting the proliferation and migration of VSMCs after vascular injury.