Identification by DNA microarray of genes involved in Candida albicans-treated gingival epithelial cells
Identification by DNA microarray of genes involved in Candida albicans-treated gingival epithelial cells
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DOI：
10.1111/j.1600-0714.2012.01149.x
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发表时间：
2012-11-01
影响因子：
3.3
通讯作者：
Abiko, Yoshimitsu
中科院分区：
文献类型：
--
作者：
Ikuta, Tonami;Bhawal, Ujjal K.;Abiko, Yoshimitsu
背景：口腔上皮细胞显著影响口咽念珠菌病中宿主对白色念珠菌的炎症反应。在口腔上皮细胞感染白色念珠菌过程中，促炎细胞因子作为早期先天免疫系统的介质起作用。我们试图阐明控制人类牙龈上皮细胞（hgec）感染白色念珠菌的分子机制模式，可能涉及多种会聚信号转导途径。材料与方法：用白色念珠菌ATCC90029培养原代hgec。感染8 h后提取总RNA，使用Affymetrix基因芯片（Human Genome U133 plus 2.0 Array， 48000个基因）监测mRNA水平。GeneChip数据采用genesspring软件和Ingenuity Pathway Analysis系统进行分析。采用逆转录聚合酶链反应（RT-PCR）、实时RT-PCR和免疫组织化学检测基因表达变化。结果：差异表达的基因代表了多种功能，如免疫反应和炎症疾病。与对照细胞相比，IL-8、ICAM-1和Cox-2的表达变化大于2倍。通过RT-PCR和real-time RT-PCR证实基因芯片分析中mRNA水平的改变。感染白色念珠菌的大鼠牙龈上皮细胞对ICAM-1和Cox-2具有较强的免疫反应性。在白色念珠菌处理的细胞中，我们发现差异基因表达随着IL-8的上调而上调或下调。结论：这些研究结果表明，白细胞介素8对白色念珠菌感染反应的分子机制可能涉及多种趋同的信号转导途径。口腔病理杂志，2012,41:769-778
BACKGROUND: Oral epithelial cells significantly influence host inflammatory responses against Candida albicans in oropharyngeal candidiasis. Pro-inflammatory cytokines function as an early innate immune system mediator during C. albicans infection in oral epithelial cells. We sought to elucidate the pattern of the molecular mechanisms governing the human gingival epithelial cells (HGECs) to C. albicans infection likely involve multiple converging signal transduction pathways.MATERIALS AND METHODS: Primary HGECs were cultured with C. albicans ATCC90029. Total RNA was extracted after 8 h of infection and monitored mRNA levels using Affymetrix GeneChip (Human Genome U133 plus 2.0 Array, 48 000 genes). GeneChip data was analyzed by GeneSpring software and Ingenuity Pathway Analysis system. Reverse transcription polymerase chain reaction (RT-PCR), real-time RT-PCR and immunohistochemistry were used to investigate gene expression changes.RESULTS: The differentially expressed genes represented functions as diverse as immune response and inflammatory disease. IL-8, ICAM-1 and Cox-2 showed a greater than two fold change in expression relative to those in control cells. Altered mRNA levels in GeneChip analysis were confirmed by RT-PCR and real-time RT-PCR. Stronger immunoreactivity against ICAM-1 and Cox-2 was also observed in the infection with C. albicans in rat gingival epithelium. We have identified differential gene expression up-regulated or down-regulated with the up-regulation of IL-8 in C. albicans-treated cells.CONCLUSION: These findings indicate that the molecular mechanisms underlying the IL-8 response of HGECs to C. albicans infection likely involve multiple converging signal transduction pathways. J Oral Pathol Med (2012) 41: 769-778