Claudin在幽门螺杆菌和AGS细胞相互作用不同时间点表达分析

The expression of Claudins in gastric epithelial cells induced by Helicobacter pylori strains at different time-points

目的研究Claudin家族成员在Helicobacter pylori(H.pylori)和AGS相互作用的不同时间点表达谱变化。方法TRIzol方法提取H.pylori26695攻击AGS细胞0、0.5、2、4、6 h时间点的细胞总RNA样品,对样品mRNA进行线性扩增后,应用IlluminaHuman-6芯片检测基因表达量,以Detection Score≥0.99;|DiffScore|≥13;方差分析(ANOVA)P≤0.05作为筛选差异基因的标准;比较Claudin家族成员的表达量。结果Claudin家族有4个成员存在差异表达,其中在早期Claudin 15下调,4 h后无差异,Claudin 2、23早期上调后于4 h和6 h分别恢复至无差异水平,而Claudin 6在0.5 h后持续上调。结论Claudin蛋白家族2、6、15、23成员参与H.pylori感染过程,并且呈现时序性变化,为H.pylori感染能引发胃癌机制研究提供线索。

Objective To investigate the expression of the Claudin family in AGS coculture with Helicobacter pylori （H. pylori）at different time-points. Methods To elucidate the sequential cross-talk between H. pylori and gastric epithelial cells, the bacteria gastric epithelial cells （AGS） were co-cultured with H. pylori 26695 strain and total RNAs were extracted with Trizol from AGS cells on 0 h, 0.5 h, 1 h, 2 h, 4 h, 6 h and used for detection of genes represented in the Illumina＇s Human-6 v2 BeadChip. The differentially expressed genes （ Dttcstion Score≥0.99 and I Diffcore ≥ 13, which equated to a nominal P-value of ≤0.05） of each time point. Results Four members of the Claudin family were differentially expressed genes, which Claudinl5 was down-regulation, and the early stage of infection Claudin 2 and Claudin 23 were up-regulation, and Claudin 6 continued upward. Conclusion The tight junction proteins called Claudins involved in infection process. The sequential gene expression profiles of Claudin 2,6,15 and 23 in gastric epithelial cells were induced by H. pylori strains-implication of mechanism for gastric carcinogenesis by H. pylori infection.