Hori Y, Nishida K, Yamato M, Sugiyama H, Soma T, Inoue T, Maeda N, Okano T, Tano Y
Department of Ophthalmology, Osaka University Medical School, Suita, Osaka 565-0871, Japan.
Exp Eye Res. 2008 Sep;87(3):191-6. doi: 10.1016/j.exer.2008.05.014. Epub 2008 Jul 21.
Cultivated oral mucosal epithelial sheet transplantation is a new surgical strategy to treat severe ocular surface disorders such as chemical burns, ocular cicatricial pemphigoid, and Stevens-Johnson syndrome. MUC16 is thought to be the most important membrane-associated mucin on the ocular surface because it forms a protective barrier on the epithelial cell surface. In this study, we studied MUC16 expression in mRNA and protein levels and compared the expression patterns between cultivated oral mucosal epithelial cell sheet and oral mucosal tissue. Specimens (5x5 mm) of oral mucosal tissue harvested from healthy volunteers were used. The oral mucosal epithelial cells were cultured on temperature-responsive culture dishes to generate stratified cell sheets. Cultivated oral mucosal epithelial cells formed three- to five-cell thick stratified sheets for 2 weeks. Scanning electron micrographs revealed that the apical surfaces of the oral mucosal tissue and the oral mucosal sheets were covered with dense microvilli/microplicae. Real-time PCR showed significantly more MUC16 transcripts in the cultivated oral mucosal sheets and corneal epithelial sheets than in the oral mucosal tissue (P=0.023 and 0.008, respectively, Mann-Whitney rank sum test). These findings were confirmed by immunohistochemical examination using an MUC16 antibody to the protein. MUC16 protein was localized to the apical cells of the oral mucosal sheets, but the human oral mucosal tissue did not express MUC16 protein in any cell layers. In this study, interestingly, the expression of membrane-associated mucin MUC16 differs between human oral mucosal epithelia and cultivated epithelial sheets. MUC16 expressed in the oral mucosal sheets may contribute to ocular surface reconstruction after oral mucosal sheet transplantation.
培养的口腔黏膜上皮片移植是一种治疗严重眼表疾病(如化学烧伤、瘢痕性类天疱疮和史蒂文斯-约翰逊综合征)的新手术策略。MUC16被认为是眼表最重要的膜相关黏蛋白,因为它在上皮细胞表面形成保护屏障。在本研究中,我们研究了MUC16在mRNA和蛋白质水平的表达,并比较了培养的口腔黏膜上皮细胞片和口腔黏膜组织之间的表达模式。使用从健康志愿者获取的口腔黏膜组织标本(5×5毫米)。将口腔黏膜上皮细胞培养在温度响应培养皿上以生成分层细胞片。培养的口腔黏膜上皮细胞在2周内形成三到五层厚的分层片。扫描电子显微镜照片显示,口腔黏膜组织和口腔黏膜片的顶端表面覆盖着密集的微绒毛/微皱襞。实时PCR显示,培养的口腔黏膜片和角膜上皮片中的MUC16转录本明显多于口腔黏膜组织(分别为P=0.023和0.008,曼-惠特尼秩和检验)。使用针对该蛋白的MUC16抗体进行免疫组织化学检查证实了这些发现。MUC16蛋白定位于口腔黏膜片的顶端细胞,但人类口腔黏膜组织在任何细胞层中均未表达MUC16蛋白。在本研究中,有趣的是,膜相关黏蛋白MUC16在人类口腔黏膜上皮和培养的上皮片之间的表达有所不同。口腔黏膜片中表达的MUC16可能有助于口腔黏膜片移植后的眼表重建。