摘要
目的:探讨S100A9在具核梭杆菌(Fusobacterium nucleatum,Fn)促结肠癌HCT116和SW480细胞增殖与迁移中的作用。方法:Fn感染HCT116和SW480细胞后,分别采用CCK8实验及Transwell实验检测结肠癌细胞的增殖及迁移能力的变化,采用Real time PCR及Western blot分别检测S100A9基因及蛋白质水平的变化;用S100A9 siRNA转染HCT116和SW480细胞后感染Fn,采用CCK8实验及Transwell实验分别检测两株细胞增殖及迁移能力的变化;利用NF-κB抑制剂BAY 11-7082预处理HCT116和SW480细胞后再感染Fn,采用Real time PCR及Western blot分别检测S100A9基因及蛋白质水平的变化。结果:(1)Fn可促进HCT116和SW480细胞增殖及迁移(P<0.001);(2)Fn感染的HCT116和SW480细胞中S100A9基因及蛋白质水平均较相应对照组明显升高,差异均具有高度显著性(P<0.01或P<0.001),即Fn上调S100A9的表达;(3)用S100A9 siRNA下调HCT116和SW480细胞中S100A9表达后,Fn促进这两株细胞增殖及迁移的作用则明显减弱,差异均具有高度显著性(P<0.001),表明S100A9参与介导Fn的促HCT116和SW480细胞增殖及迁移的作用;(4)在NF-κB通路抑制剂BAY 11-7082预处理的HCT116和SW480细胞组,Fn上调S100A9基因及蛋白质水平的作用明显被逆转,差异均具有高度显著性(P<0.01或P<0.001),表明激活NF-κB转录活性是Fn上调S100A9的机制之一。结论:Fn可上调S100A9表达且与NF-κB活化有关,上调S100A9是Fn促结肠癌细胞增殖及迁移的机制之一。
Objective:To investigate the effect of S100A9 in proliferation and migration in Fusobacterium nucleatum(Fn)-induced colon cancer HCT116 and SW480 cells.Method:CCK8 assay and Transwell assays were used to detect the proliferation and migration of colon cancer HCT116 and SW480 cells infected with Fn.Real time PCR and Western blot were used to detect expression of S100A9 mRNA and protein in HCT116 and SW480 cells infected with Fn,respectively.After transfecting S100A9 siRNA into cells for silencing S100A9 expression,CCK8 assay and Transwell assays were used to detect the changes of proliferation and migration of HCT116 and SW480 cells infected with Fn.After pre-treatment with NF-κB inhibitor BAY 11-7082,Real time PCR and Western blot were used to detect the changes of S100A9 mRNA and protein in HCT116 and SW480 cells infected with Fn.Result:(1)Fn promotes the proliferation and migration of HCT116 and SW480 cells(P<0.001).(2)The levels of S100A9 mRNA and protein in Fn-infected HCT116 and SW480 cells were significantly higher than that in the control group(P<0.01 or P<0.001),suggesting that S100A9 expression is up-regulated by Fn.(3)The proliferation and migration of Fn-infected HCT116 and SW480 cells were e rescued after silencing S100A9 expression(P<0.001),indicating that S100A9 is involved in it.(4)NF-κB inhibitor effectively inhibited the increase of S100A9 expression induced by Fn.Conclusion:Fn can regulate S100A9 expression by affecting NF-κB activation,and S100A9 is involved in Fn-mediated proliferation and migration of colon cancer cells.
作者
户丽君
段亮
黄逸云
林璐
黄茂
陈露
彭棋
胡琴
张彦
周兰
HU Li-jun;DUAN Liang;HUANG Yi-yun;LIN Lu;HUANG Mao;CHEN Lu;PENG Qi;HU Qin;ZHANG Yan;ZHOU Lan(Key Laboratory of Laboratory Medical Diagnostics of Ministry of Education,Chongqing Medical University,Chongqing 400016,China;Department of Laboratory Medicine,The Second Affiliated Hospital of Chongqing Medical University,Chongqing 400010,China)
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2020年第1期84-91,共8页
China Biotechnology
基金
国家自然科学基金(81601837)资助项目.
