Paper For Master Academic Degree The Effects And Mechanisms Of Tetraspanin CD151-ARSA Mutant In Angiogenesis

Purpose: The purpose of this study was to assess whether CD151-ARSA by PI3K/AKt,ERK signaling pathways the effects and mechanisms angiogenesis in human umbilical vein endothelial cells (HUVEC) and in vitro.Methods: 1.CD151 and CD151-ARSA mutants (interfering vesicular trafficking) were inserted into recombinant adeno-associated virus (rAAV) vectors and used to transfect HUVECs. After transfection, the expression of CD151-ARSA was measured. 2.Proliferation was assessed using the Cell Counting Kit-8 assay, 3.cell migration was evaluated by wound healing assay, 4.the tube formation assay was performed on Matrigel. 5.The potential involvement of important signaling pathways was explored by the PI3K inhibitor (LY294002, 15μM) and MAPK inhibitor (apigenin, 25μM) and further explore the CD151-ARSA effects and mechanisms angiogenesis in vitro.Results: 1.CD151-ARSA gene delivery increased the expression of CD151 at the protein level. 2.CD151-ARSA compared with CD151 decreased cell proliferation, migration and tube formation . 3. CD151-ARSA affect angiogenesis involved by the PI3K /AKt,ERK signaling pathways. Conclusions: This study demonstrates;1.Confirmed CD151-ARSA mutant affected angiogenesis in HUVECs .2.Confirmed that the CD151-ARSA affected angiogenesis,but the CD151-ARSA compared with CD151 decreasd effects of activity .3.PI3K and ERK inhibtor could inhibit CD151-ARSA effects in angiogenesis,proved that it is through the PI3K/AKt,ERK signaling pathways to affect angiogenesis. Our observations provide a new insight in the mechanism of CD151 as a novel therapeutic strategy for regulating angiogenesis.