Transcriptome Profiles of Human Lung Epithelial Cells A549 Interacting with Aspergillus fumigatus by RNA-Seq.
Transcriptome Profiles of Human Lung Epithelial Cells A549 Interacting with Aspergillus fumigatus by RNA-Seq.
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通过 RNA-Seq 分析人肺上皮细胞 A549 与烟曲霉相互作用的转录组图谱
DOI：
10.1371/journal.pone.0135720
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Han L
中科院分区：
文献类型：
--
作者：
Chen F;Zhang C;Jia X;Wang S;Wang J;Chen Y;Zhao J;Tian S;Han X;Han L
肺上皮细胞构成了宿主抵抗烟曲霉吸入的第一道防线，但人肺泡II型上皮细胞的转录应答机制尚不清楚。在这里，我们使用RNA-Seq技术来评估A549细胞与A.烟熏。鉴定的基因总数为19118个。与未感染的A549细胞相比，分生孢子共培养8 h的细胞中有459个基因表达差异，其中上调基因302个，下调基因157个。GO和KEGG途径富集分析显示，上调表达的基因大多与免疫反应、趋化性和炎症反应有关，并在嘌呤-细胞因子受体相互作用、JAK-STAT和MAPK信号通路中富集。下调的基因主要集中在与发育、造血和离子转运相关的基因。其中，EGR 4和HIST 1H 4J基因分别是上调和下调基因中倍数变化最大的。进一步验证了14个上调基因和3个下调基因，qRT-PCR证实A549细胞与A.烟熏。Western blot结果显示，在A549细胞与A.烟曲霉分生孢子培养8h。干扰A549细胞内源性ARC或EGR 1蛋白表达可降低A.烟熏。这些结果为了解肺上皮细胞基因表达的动态变化，特别是肺上皮细胞对A.烟曲霉感染
Lung epithelial cells constitute the first defense line of host against the inhaled Aspergillus fumigatus; however, the transcriptional response of human alveolar type II epithelial cells was still unclear. Here we used RNA-Seq technology to assess the transcriptome profiles of A549 cells following direct interaction with conidia of A. fumigatus. The total number of identified genes was 19118. Compared with uninfected A549 cells, 459 genes were differentially expressed in cells co-incubated with conidia for 8 h, including 302 up-regulated genes and 157 down-regulated genes. GO and KEGG pathway enrichment analysis showed that most of the up-regulated genes were related to immune response, chemotaxis and inflammatory response and enriched in cytokine-cytokine receptor interaction, JAK-STAT and MAPK signaling pathways. The down-regulated genes were mainly enriched for terms associated with development, hemopoiesis and ion transport. Among them, EGR4 and HIST1H4J gene had the maximum of fold change in up-regulated and down-regulated genes, respectively. Fourteen up-regulated genes and three down-regulated genes were further validated and significant increase on expression of IL-6, IL-8 and TNF-α in A549 cells were confirmed by qRT-PCR during the interaction of A549 cells with A. fumigatus. Besides, western blot showed that expression of two proteins (ARC, EGR1) significantly increased in A549 cells during interaction with A. fumigatus conidia for 8h. Interference of endogenous expression of ARC or EGR1 protein in A549 cells reduced the internalization of A. fumigatus. These results provided important insights into dynamic changes of gene expression in lung epithelial cells, especially its strong immunological response against A. fumigatus infection.