弓形虫多态性ROP16效应分子诱导宿主A549细胞基因表达谱的差异研究

目的 探讨弓形虫Ι、Ⅱ和Ⅲ型ROP16对宿主基因表达谱的影响及其意义。方法 以A549细胞株为研究模型,建立弓形虫Ι、Ⅱ和Ⅲ型ROP16稳定转染的A549细胞株,通过表达谱芯片技术筛选差异表达基因,并对差异基因进行GO分析和Pathway通路分析。结果 对差异基因进行聚类,发现ROP16_I转染组776个基因表达上调,494个基因表达下调;ROP16_III转染组842个基因表达上调,520个基因表达下调;而ROP16_II转染组5 063个基因表达上调,5 989个基因表达下调。另外,ROP16_I/III转染组基因表达谱相似。而ROP16_II转染组则不同,表达谱差异显著。对ROP16_II转染组差异基因进行Pathway通路分析发现,显著改变的通路共有74条,其中ROP16_II单独调控的信号通路有19条,主要与NF-κB、Toll样受体,趋化因子等信号通路、炎症反应及免疫调节和代谢等相关。结论 ROP16入侵宿主细胞核后影响宿主细胞基因表达谱。同Ι、Ⅲ型弓形虫相比,Ⅱ型弓形虫在基因组上的差异导致其具有独特的差异基因表达谱和信号通路,了解其单独调控的信号通路对于Ⅱ型弓形虫的防治具有重要意义。

Differential expression profiling of A549 cells induced by Toxoplasma polymorphism effector ROP16

The aim of this study is to explore the changes and significance of gene expression profile induced by ROPl6 of Toxoplasma gondii (T. gondii) type I, II, III. The A549 cell line was used as the research model. And, stable transfected ROPl6 of type I, II, III of T. gondii cell lines were established. Then, we used gene expression microarray to identify genes that were differentially expressed. Gene Ontology (GO) functional classification and pathway analysis were performed on screened differentially expressed genes. According to the results of the gene expression profile, ROP16_I transfer group owned 1 270 different genes, with 776 genes up-regulated and 494 genes down-regulated; ROP16_III transfer group owned 1 362 different genes, with 842 genes up-regulated and 520 genes down-regulated; ROP16_II transfer group owned 11 052 different genes, with 5 063 genes up-regulated and 5 989 genes down-regulated. The ROP16_I/III group has the similar expression spectrum. However, ROP16_II transfer group was very different, which has distinct molecular profiles. Pathway analysis showed 74 pathway changes of the ROP16_II transfer group, and 19 pathways was solely dominated by it. In addition, these pathways were related to NF-kappa B, Toll-like receptor, Chemokine signaling pathway and inflammatory response, immune response and metabolic process. Above all, ROP16 could affect host cell gene expression profiles. And, compared to Toxoplasma gondii type I, III, type II has unique gene expression profiles and signaling pathway. Understanding its unique signaling pathway is very important for the prevention of type II T. gondii.