Toll-like receptor 2-mediated gene expression in epithelial cells during Helicobacter pylori infection
Toll-like receptor 2-mediated gene expression in epithelial cells during Helicobacter pylori infection
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DOI：
10.1111/j.1523-5378.2005.00311.x
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发表时间：
2005-06-01
期刊：
影响因子：
4.4
通讯作者：
Goldberg, JB
中科院分区：
文献类型：
--
作者：
Ding, SZ;Torok, AM;Goldberg, JB
背景。幽门螺杆菌是引起慢性胃炎和消化性溃疡病的主要病原菌，与胃恶性肿瘤密切相关。我们之前已经证明，幽门螺杆菌诱导的 NF-κ B 激活和白细胞介素 (IL)-8 分泌是由上皮细胞中的 Toll 样受体 (TLR) 2 介导的。然而，幽门螺杆菌感染期间TLR2介导的上皮细胞整体基因表达谱仍不清楚。本研究的目的是鉴定幽门螺杆菌诱导的上皮细胞中 TLR2 调节基因。材料和方法。 HEK293和HEK-TLR2细胞与幽门螺杆菌26695共培养6小时。提取总 RNA 并与 Affymetrix 人类 U133A 微阵列芯片组杂交，该芯片组包含 22,283 个总探针组，其中包括 14,285 个基因。使用 AFFYMETRIX SUITE 5 软件进行数据分析。通过定量逆转录酶-聚合酶链反应（RT-PCR）监测胃上皮细胞AGS和MKN45中选定基因的表达。结果。 HEK293 细胞感染幽门螺杆菌后，57 个探针组中包含的 46 个基因被诱导 > 2 倍，3 个基因（5 个探针组）减少 > 2 倍。在幽门螺杆菌感染的 HEK-TLR2 细胞中，69 个探针组中包含的 54 个基因被诱导 > 2 倍，而只有 1 个基因被抑制 > 2 倍。对 HEK293 或 HEK-TLR2 细胞中诱导的基因进行比较，确定了 28 个基因，其表达依赖于 TLR2 的存在。选择了 17 个基因，并在幽门螺杆菌感染期间使用定量 RT-PCR 评估了胃上皮细胞中的表达。幽门螺杆菌刺激后，17 个基因中的 8 个在 AGS 和 MKN45 细胞中显示出不同的表达模式。结论。目前的研究调查了上皮细胞系统中幽门螺杆菌刺激后 TLR2 介导的全局基因变化。这种方法将有助于鉴定其表达由特定 TLR 介导的基因，并有助于确定幽门螺杆菌感染期间负责不同信号途径的细胞反应。
Background. Helicobacter pylori is the major pathogen causing chronic gastritis and peptic ulcer disease and is closely linked to gastric malignancy. We have previously shown that H. pylori-induced NF-kappa B activation and interleukin (IL)-8 secretion are mediated by Toll-like receptor (TLR) 2 in epithelial cells. However, the TLR2-mediated global gene expression profile of the epithelial cell during H. pylori infection is still unknown. The goal of this study was to identify TLR2-regulated genes in epithelial cells induced by H. pylori.Materials and methods. The HEK293 and HEK-TLR2 cells were cocultured with H. pylori 26695 for 6 hours. Total RNA was extracted and hybridized to the Affymetrix human U133A microarray chipset, which contains 22,283 total probe sets including 14,285 genes. Data analyses were performed using AFFYMETRIX SUITE 5 software. The expression of selected genes in gastric epithelial cells AGS and MKN45 was monitored by quantitative reverse transcriptase-polymerase chain reaction (RT-PCR).Results. Forty-six genes, contained in 57 probe sets, were induced > 2-fold and three genes (five probe sets) decreased > 2-fold by H. pylori infection of HEK293 cells. Fifty-four genes, contained in 69 probe sets, were induced > 2-fold, whereas only 1 gene was repressed > 2-fold in H. pylori-infected HEK-TLR2 cells. Comparisons of genes induced in HEK293 or HEK-TLR2 cells identified 28 genes whose expression was dependent on the presence of TLR2. Seventeen genes were selected and their expression was assessed using the quantitative RT-PCR in gastric epithelial cells during H. pylori infection. Eight of the 17 genes showed distinct expression patterns in AGS and MKN45 cells after H. pylori stimulation.Conclusions. The current study investigated the TLR2-mediated global gene changes after H. pylori stimulation in the epithelial cell system. This approach will be helpful in identifying genes whose expression is mediated by specific TLRs and in determining the cellular responses that are responsible for diverse signal pathways during H. pylori infection.