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作 者:杨五小[1] 林敬阳[1] 刘正湘[1] 秦瑾[1] 左后娟[1] 费宇杰[1] 彭丹[1]
机构地区:[1]华中科技大学同济医学院附属同济医院心内科,邮政编码武汉430030
出 处:《微循环学杂志》2009年第4期5-8,共4页Chinese Journal of Microcirculation
基 金:国家自然科学基金资助项目(NO:30670856)
摘 要:目的:研究CD151-整合素α3/α6复合体对人脐静脉内皮细胞(HUVEC)增殖的影响及机制。方法:将重组腺相关病毒介导CD151基因(rAAV-CD151)和整合素α3/α6结合缺陷的突变CD151基因(rAAV-CD151-AAA194-196)转染HUVEC。免疫共沉淀检测CD151-整合素α3/α6复合体;Western blot检测CD151、磷脂酰肌醇-3激酶(PI3K)、丝/苏氨酸蛋白激酶B(Akt)、磷酸化Akt(p-Akt)的表达;MTT法检测CD151对HUVEC增殖的影响。结果:CD151组和CD151突变组的CD151蛋白表达水平较对照组、绿色萤光蛋白组(GFP组)明显增加(P<0.05)。CD151组的CD151-整合素α3/α6复合体、PI3K和p-Akt表达水平明显高于对照组和GFP组(P<0.05);CD151突变组的CD151-整合素α3/α6复合体、PI3K和p-Akt表达水平明显低于对照组和GFP组(P<0.05)。CD151组的细胞增殖明显高于对照组和GFP组,CD151突变组的细胞增殖能力最低。结论:过度表达CD151蛋白可以促进HUVEC的增殖,其机制是过度表达的CD151分子与整合素形成复合体增加,使PI3K和p-Akt表达增加。Objective:To investigate the the effect of CD151 integrin α3/α6 complex on proliferation of human umbilical vein endothelial cells (HUVEC) and the mechanism involved. Method: Recombinant adeno-associated virus mediated normal CD151 gene (rAAV-CD151) or mutant CD151 gene (rAAV-CD151-AAA194-196) which ability of combinat with integrin α3/α6 was weaken,delivery HUVEC. Then HUVEC were assigned to control group,GFP group,CD151 group,mutant CD151 group. After gene delivery HUVEC,the expression of CD151and PI3K,Akt were measured by Western blot. To study the capacity of CD151 form complex with the integrin α3/α6 through the method of co-immunoprecipitation. Cell proliferation assay was evaluated by MTT. Results: The expression of CD151 protein was increased in mutant CD151 group and CD151 group,compared with the group of GFP and control. Phosphorylation of Akt,PI3K,CD151-integrin α3/α6 complex in CD151 group were much higher,compared with the control group and GFP group. The expression of PI3K,phosphorylation of Akt,CD151-integrin α3/α6 complex of mutant CD151 group were markedly reduced,compared with the control group and the GFP group. The abilty of cell proliferation was much higher in CD151 group,compared with the control group and the GFP group (P<0.05). The abilty of cell proliferation in mutant CD151 group was the minimum of the all four groups. Conclusion: Normal CD151 gene delivery promotes the proliferation of HUVEC. The molecular mechanism involved may be that over-expression of CD151 promotes CD151 interact with integrin α3/α6 and increase the expression of CD151-integrin α3/α6 complex and result to up-regulated the PI3K and phosphorylation of Akt.
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