摘要
目的分析幽门螺杆菌分泌的细胞毒素相关蛋白A(CagA)对胃黏膜Runx3抑制作用的分子机制。方法构建CagA表达载体pWT-CagA,突变载体PR-CagA(B-)、PR-Cag(D-),利用上述载体及空白载体转染人胃黏膜上皮永生化细胞GES-1,分别标记为pWT-CagA质粒组、PR-CagA(B-)质粒组、5 PR-CagA(D-)质粒组、空载体组,另设空白对照组。采用免疫印迹方法检测Runx3蛋白表达水平。应用BAY11-7082、PP1、PD98059、SB203580预孵育GES-1细胞,再利用pWT-CagA质粒及空转染质粒转染GES-1细胞,探讨CagA对Runx3表达影响的分子机制。结果pWT-CagA转染后,GES-1细胞Runx3蛋白表达量明显低于空白对照组、空转染组及PR-CagA(B-)转染组、PR-Cag(D-)转染组。应用PP1、PD98059、SB203580预孵育GES-1细胞后,pWT-CagA组和空载体组Runx3表达水平对比,差异无统计学意义(P>0.05)。应用BAY11-7082预孵育GES-1细胞后,pWT-CagA组Runx3表达水平仍明显低于空载体组,差异有统计学意义(P<0.05)。结论幽门螺杆菌CagA能够抑制胃黏膜Runx3的表达,且这种抑制作用可能与MAPK/MEK/ERK信号通路有关。
Objective To analyze the molecular mechanism of cytotoxin-associated protein A(CagA)secreted by Helicobacter pylori in inhibiting Runx3 in gastric mucosa.Methods CagA expression vectors pWT-CagA,CagA mutation vectors PR-CagA(B-),PR-Cag(D-)were constructed.The above vectors and blank vectors were used to transfect GES-1,respectively.They were labelled as PWT CagA plasmid group,PR CagA(B-)plasmid group,5 PR CagA(D-)plasmid group,empty body group,another blank control group was established,too.The expression of Runx3 protein was detected by immunoblotting.GES-1 cells were pre-incubated with BAY11-7082,PP1,PD98059 or SB203580,and then transfected with pWT-CagA plasmid and empty transfection plasmid to explore the molecular mechanism of the effect of CagA on Runx3 expression.Results After pWT-CagA transfection,the expression of Runx3 protein in GES-1 cells was significantly lower than that in blank control group,blank transfection group,PR-CagA(B-)transfection group and PR-Cag(D-)transfection group.After incubation with PP1,PD98059 and SB203580,there was no significant difference in Runx3 expression between pWT-CagA group and empty vector group(P>0.05).After pre-incubation with BAY11-7082,the expression of Runx3 in pWT-CagA group was significantly lower than that in empty vector group(P<0.05).Conclusion Helicobacter pylori CagA can inhibit the expression of Runx3 in gastric mucosa,and this inhibition may be related to the MAPK/MEK/ERK signaling pathway.
作者
许燕川
符丽珍
何利沉
XU Yan-chuan;FU Li-zhen;HE Li-zhen(Hainan Blood Center,Haikou,Hainan,570311,China;Department of Gastroenterology,the Second Affiliated Hospital of Hainan Medical College,Haikou,Hainan,570216,China)
出处
《现代消化及介入诊疗》
2019年第11期1270-1274,共5页
Modern Interventional Diagnosis and Treatment in Gastroenterology
