溶血磷脂酸对上皮细胞整合素β_6表达的影响  

Effect of lysophosphatidic acid on expression of integrin β_6 in epithelial cells

作　　者：许铭炎[1] 邓小玲[2] 陈锡和[3] 杨利和[1] 姚小武[1] 傅玉才[3] 

机构地区：[1]汕头大学医学院第二附属医院口腔科,广东汕头515041 [2]汕头大学医学院细胞生物与遗传学教研室,广东汕头515041 [3]汕头大学医学院细胞衰老实验室,广东汕头515041

出　　处：《中国病理生理杂志》2012年第3期488-491,共4页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81072208);广东省自然科学基金资助项目(No.S2011010005106);广东省医学科研基金资助项目(No.A2010407);汕头大学医学院基础与临床科研基金资助项目

摘　　要：目的:探讨溶血磷脂酸(LPA)在体外对整合素β_6(ITGB6)表达的影响,并进一步研究LPA诱导的活化转化生长因子β(TGF-β)在此过程中的作用。方法:原代培养的正常人支气管上皮(NHBE)细胞接种于6孔板中,经LPA诱导,收集细胞,分别利用RT-PCR和流式细胞术检测ITGB6 mRNA及细胞表面蛋白的表达;利用转染有TGF-β应答性纤溶酶原激活物抑制剂1(PAI-1)启动子片段并融合萤火虫荧光报告基因片段的转化貂肺上皮细胞(TMLC)作为TGF-β活性报告细胞检测活化的TGF-β。结果:(1)10μmol/L LPA诱导2 h后,ITGB6mRNA在上皮细胞中表达显著增加,具有明显的时间依赖性。(2)10μmmol/L LPA诱导4 h后,上皮细胞表面ITGB6蛋白表达明显增加。(3)抗α_vβ_6抗体可阻断LPA诱导的活化TGF-β,但不能阻断LPA诱导的ITGB6 mRNA的表达。结论:(1)LPA可诱导上皮细胞ITGB6 mRNA和细胞表面蛋白的表达。(2)LPA诱导的ITGB6 mRNA表达不依赖LPA诱导的TGF-β活化。AIM：To investigate the effect of lysophosphatidic acid（LPA） on the expression of integrin B_6（ITGB6） for determining the role of transforming growth factor B（TGF-B） activation induced by LPA in this process. METHODS：Normal human bronchial epithelial（NHBE） cells were primarily cultured in 6 well plate and stimulated with LPA.The mRNA expression of ITGB6 and the level of cell surface ITGB6 protein were detected by RT-PCR and flow cytometry, respectively.The activity of active TGF -βinduced by LPA was measured by the method of transformed mink lung epithelial cells（TMLC） transfected with TGF -βresponsive plasminogen activator inhibitor 1（PAI-1 ） promoter fused with firefly luciferase reporter gene.RESULTS：After stimulated with LPA at concentration of 10μmol/L for 2 h,the mRNA expression of ITGB6 in epithelial cells was significantly increased in a time-dependent manner.The results of flow cytometry showed that the protein level of ITGB6 on cell surface was obviously increased after treated with LPA at concentration of 10μmol/L for 4 h.The active TGF -βinduced by LPA in epithelial cells was blocked by anαvB6 blocking antibody. However,α_vP_6 blocking antibody failed to inhibit the mRNA expression of ITGB6 induced by LPA.CONCLUSION： LPA induces the mRNA and cell surface protein of ITGB6 in epithelial cells.The up-regulated ITGB6 expression by LPA is independent on LPA-induced TGF -βactivation.

关 键 词：溶血磷脂酸 整合素β6 上皮细胞 转化生长因子Β 

分 类 号：R735.35[医药卫生—肿瘤]