Kim Chang-Hoon, Kim Hyun Jik, Song Kyoung Seob, Seong Je-Kyung, Kim Kyung-Su, Lee Jeung-Gweon, Yoon Joo-Heon
Department of Otorhinolaryngology, Yonsei University College of Medicine, Seoul, South Korea.
Acta Otolaryngol. 2005 Jan;125(1):76-81. doi: 10.1080/00016480410015785.
This study indicates that MUC8 protein is expressed in ciliated cells from human nasal epithelial cells and is upregulated by IL-1beta treatment. These results suggest that MUC8 gene and protein expression levels could be used as a ciliated cell marker in human nasal epithelium.
To examine MUC8 mRNA expression patterns according to the mucociliary differentiation of normal human nasal epithelial (NHNE) cells, and to investigate the localization of MUC8 proteins in nasal polyps.
Passage-2 NHNE cells were cultured using an air-liquid interface technique. On Days 2, 7, 14 and 28 after confluence, ciliated cells were counted by means of cytospin slide immunostaining using H6C5 and beta-tubulin, and MUC8 mRNA levels were determined using real-time quantitative polymerase chain reaction (PCR). After synthesizing polyclonal anti-MUC8 peptide antibodies, MUC8 immunostaining was performed using nasal polyps. MUC8 mRNA and protein levels were determined in NHNE cells treated with IL-1beta (10 ng/ml for 24 h) using reverse transcriptase-PCR and Western blot analysis.
The increases in the number of ciliated cells and the MUC8 gene expression level with increasing culture time in the NHNE cells were quite similar. MUC8 was expressed in the ciliated cells of human nasal polyps. The MUC8 protein level and the mRNA level were upregulated as a result of IL-1beta treatment.
本研究表明,MUC8蛋白在人鼻上皮细胞的纤毛细胞中表达,并在白细胞介素-1β处理后上调。这些结果表明,MUC8基因和蛋白表达水平可作为人鼻上皮中纤毛细胞的标志物。
根据正常人鼻上皮(NHNE)细胞的黏液纤毛分化情况,检测MUC8 mRNA的表达模式,并研究MUC8蛋白在鼻息肉中的定位。
使用气液界面技术培养第2代NHNE细胞。汇合后第2、7、14和28天,通过使用H6C5和β-微管蛋白的细胞涂片免疫染色对纤毛细胞进行计数,并使用实时定量聚合酶链反应(PCR)测定MUC8 mRNA水平。合成多克隆抗MUC8肽抗体后,对鼻息肉进行MUC8免疫染色。使用逆转录PCR和蛋白质印迹分析测定用白细胞介素-1β(10 ng/ml,处理24小时)处理的NHNE细胞中的MUC8 mRNA和蛋白水平。
NHNE细胞中纤毛细胞数量和MUC8基因表达水平随培养时间的增加而增加,二者变化非常相似。MUC8在人鼻息肉的纤毛细胞中表达。白细胞介素-1β处理导致MUC8蛋白水平和mRNA水平上调。