具核梭杆菌通过调控miR-181b在结肠癌细胞中形成炎性微环境的机制
Chinese Journal of Gastroenterology(2017)
Abstract
背景:具核梭杆菌(Fn)是口腔常见的致病菌,研究发现Fn与结直肠癌的发生、发展密切相关,尤其是炎症相关性结直肠癌.目的:探讨Fn感染在结肠癌细胞中形成炎性微环境的机制.方法:构建Fn感染Caco-2细胞的炎症模型,行miRNA测序.将miR-181b mimics或inhibitor转染Fn感染的Caco-2细胞.以qRT-PCR和蛋白质印迹法分别检测TNF-αmRNA和蛋白表达,ELISA法检测上清液中TNF-α 含量,Transwell小室法检测穿膜淋巴细胞数.结果:Fn组TNF-αmRNA和蛋白表达均显著高于对照组(P<0.05),上清液中TNF-α含量显著升高(P<0.05),穿膜淋巴细胞数量明显增多(P<0.05).miRNA测序和qRT-PCR结果均显示,Fn组miR-181b表达较对照组显著降低(P<0.05).与对照组相比,miR-181b mimics+Fn组TNF-αmRNA和蛋白表达显著降低(P<0.05);而miR-181b inhibitor组TNF-αmRNA和蛋白表达均显著升高(P<0.05).生物信息学工具和双荧光素酶检测证实TNF-α可能为Caco-2细胞中miR-181b的靶基因.结论:Fn通过抑制Caco-2细胞中miR-181b表达而上调TNF-α表达,募集淋巴细胞形成炎性微环境.
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Key words
Fusobacterium nucleatum,miR-181b,Tumor Necrosis Factor-alpha,Colorectal Neoplasms
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