山东医药2017，Vol.57Issue(23)：1-3,3.DOI:10.3969/j.issn.1002-266X.2017.23.001
对头颈鳞状细胞癌中高表达基质金属蛋白酶基因具有激活作用的人类膜蛋白基因筛选
Screening of human membrane protein genes activating MMPs highly expressed in squamous-cell carcinoma of head and neck
摘要
Abstract
Objective To screen out the human membrane protein genes activating matrix metalloproteinases (MMPs) (MMP1, MMP2, MMP3, MMP9, MMP10, MMP12, and MMP13) highly expressed in squamous-cell carcinoma of head and neck.Methods Seven alternative MMP promoter reporter gene plasmids were constructed.PMA, a kind of protein kinase C activator, was used to stimulate them;Dual luciferase assay system was used to detect the basic and stimulated activities of MMP promoter reporters.The relative luciferase activity (RLA) values were measured as the ratio of the basic divided by the stimulated ones;the highest RLA was selected MMP promoter reporter gene plasmid.A total of 722 candidate genes encoding potential human transmembrane proteins were screened in a dual luciferase based assay system.The candidate genes with RLA greater than 10 were considered as the activator to MMP promoter reporters in the first round, and the second and third rounds followed the same way.Finally, the efficacious human membrane protein genes were ascertained.Results The RLAs of MMP1, MMP2, MMP3, MMP9, MMP10, MMP12, and MMP13 promoter reporters stimulated by PMA were 4.53±0.72, 1.47±0.28, 23.28±1.96, 2.37±0.46, 8.81±1.35, 2.15±0.51, and 3.47±0.99.The MMP3 reporter was selected for a high through-put screening for the sensitivity (P<0.05).G protein-coupled receptor 65 (GPR65), AXL receptor tyrosine kinase (AXL), and tumor necrosis factor receptor superfamily 10b (TNFRSF10B) were confirmed as significant activators to MMPs by screening all 722 plasmids in 3 rounds.Conclusion GPR65, AXL, and TNFRSF10B could activate MMPs highly expressed in head and neck squamous-cell carcinoma.关键词
头颈部鳞状细胞癌/基质金属蛋白酶/人类膜蛋白/G蛋白耦联受体65/AXL受体酪氨酸激酶/肿瘤坏死因子受体超家族成员10bKey words
head and neck squamous-cell carcinoma/matrix metalloproteinases/human membrane protein/G protein-coupled receptor 65/AXL receptor tyrosine kinase/tumor necrosis factor receptor superfamily 10b分类
医药卫生引用本文复制引用
许洪波,米光熙,黄俊伟,刘泽阳,尹高菲,黄志刚..对头颈鳞状细胞癌中高表达基质金属蛋白酶基因具有激活作用的人类膜蛋白基因筛选[J].山东医药,2017,57(23):1-3,3.基金项目
国家自然科学基金项目(81670946) （81670946）
北京市医管局临床医学发展专项扬帆计划项目(XMLX201507) （XMLX201507）
首都医科大学附属北京同仁医院科研骨干基金项目(2015-YJJ-GGL-002). （2015-YJJ-GGL-002）
