Department of Biochemistry and Molecular Biology, Nihon University School of Dentistry at Matsudo, Matsudo, Japan.
J Oral Pathol Med. 2012 Nov;41(10):769-78. doi: 10.1111/j.1600-0714.2012.01149.x. Epub 2012 May 3.
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.
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.
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.
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.
口腔上皮细胞在口腔念珠菌病中显著影响宿主对白念珠菌的炎症反应。促炎细胞因子在口腔上皮细胞对白念珠菌感染中作为早期固有免疫系统介质发挥作用。我们试图阐明调控人牙龈上皮细胞（HGEC）对念珠菌感染的分子机制模式，可能涉及多个趋同的信号转导途径。
用 C. albicans ATCC90029 培养原代 HGEC。感染 8 小时后提取总 RNA，并使用 Affymetrix GeneChip（Human Genome U133 plus 2.0 Array，48000 个基因）监测 mRNA 水平。GeneSpring 软件和 Ingenuity Pathway Analysis 系统分析 GeneChip 数据。逆转录聚合酶链反应（RT-PCR）、实时 RT-PCR 和免疫组织化学用于研究基因表达变化。
差异表达的基因代表了免疫反应和炎症性疾病等多种功能。与对照细胞相比，IL-8、ICAM-1 和 Cox-2 的表达变化大于两倍。基因芯片分析中的改变的 mRNA 水平通过 RT-PCR 和实时 RT-PCR 得到证实。在白色念珠菌感染大鼠牙龈上皮组织中也观察到 ICAM-1 和 Cox-2 的免疫反应性增强。我们已经确定了差异基因表达，与念珠菌处理细胞中 IL-8 的上调相关的基因表达上调或下调。
这些发现表明，HGEC 对念珠菌感染的 IL-8 反应的分子机制可能涉及多个趋同的信号转导途径。
