| 刘真1 贾赤宇1 肖斌2 唐彬2 黎伯胜2 毛旭虎2 邹全明2△.幽门螺杆菌感染细胞模型的建立及感染相关microRNAs 的筛选鉴定[J].现代生物医学进展英文版,2011,11(1):18-21. |
| 幽门螺杆菌感染细胞模型的建立及感染相关microRNAs 的筛选鉴定 |
| Establishment of helicobacter pylori infection cell model and identificationof infection-associated microRNAs |
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| DOI： |
| 中文关键词: 幽门螺杆菌 胃上皮细胞 microRNAs 感染 |
| 英文关键词: Helicobacter pylori Gastric epithelium cells MicroRNAs Infection |
| 基金项目:国家自然科学基金（30770113） |
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| 中文摘要: |
| 目的：建立稳定的幽门螺杆菌（H. pylori）感染人胃上皮细胞模型；筛选并鉴定H. pylori 感染相关microRNAs（miRNAs）的表
达，为深入研究感染相关miRNAs 的调控作用机制奠定基础。方法：将H. pylori 标准株按MOI=100:1 感染人胃上皮细胞，通过检
测炎性细胞因子及炎症反应关键酶的表达综合评价感染模型；采用博奥公司miRNAs V3.0 芯片分析细胞感染前后miRNAs 表达
谱变化，运用实时定量PCR 技术和Northern 杂交对表达显著差异的miRNAs 进行分析鉴定。结果：H. pylori 感染细胞24 h 后，细
胞分泌促炎细胞因子IL-8 显著升高(P<0.01)；启动炎症反应的关键酶COX-2 的表达明显增加。芯片数据显示：H. pylori 感染引起
超过2 倍显著差异表达的miRNAs 包括：表达上调的PREDICTED-MIR191、miR-155、miR-92b、miR-30b、miR-146a、miR-16 等，和
表达降低的miR-181b、miR-324。实时定量PCR 和Northern 杂交结果显示感染相关miR-155 和miR-146a 在H. pylori 感染细胞模
型中表达均显著增加(P<0.01)。结论：miR-155 和miR-146a 在感染细胞模型中的表达增加提示二者可能参与H. pylori 感染的免疫
调控过程。 |
| 英文摘要: |
| Objective: To establish stable gastric epithelium cells model infected by Helicobacter pylori (H. pylori), and to screen
and identify H. pylori infection- related microRNAs (miRNAs) so as to investigate the regulatory mechanism of miRNAs. Methods: H.
pylori were added to cells at a multiplicity of infection of 100:1. The infection model was monitored by the release of interleukin-8(IL-8)
and the expression of COX-2. The expression profile of cellular miRNAs during H. pylori infection were analyzed by microarray (CapitalBio,
Beijing). The significantly altered miRNAs were identified by the quantitative RT-PCR and Northern blot assays. Results: After
the incubation of H. pylori with the gastric epithelium cells for 24h, IL-8 release and COX-2 protein expression in cells were detected.
The expression of miRNAs could be significantly altered during H. pylori infection by microarray, including the up-regulation of PREDICTED-
MIR191, miR-155, miR-92b, miR-30b, miR-146a, miR-16 and the down-regulation of miR-181b and miR-324. In consensus
with the findings from microarray, miR-155 and miR-146a in infection models increased significantly (P<0.01). Conclusion: The upregulation
of miR-155 and miR-146a in infection models implied that they may be involved in the immune regulation of H. pylori infection. |
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