Grimm Martin, Renz Cornelius, Munz Adelheid, Hoefert Sebastian, Krimmel Michael, Reinert Siegmar
Department of Oral and Maxillofacial Surgery, University Hospital Tübingen, Osianderstrasse 2-8, 72076, Tübingen, Germany,
Odontology. 2015 Jan;103(1):36-49. doi: 10.1007/s10266-013-0133-2. Epub 2013 Aug 25.
Receptor activator of nuclear factor-kappa (RANK)/receptor activator of nuclear factor-kappa B ligand (RANKL) signaling helps putative cancer stem cells (CSC) to maintain their stemness. Expression of CD44 and RANKL was analyzed in oral squamous cell carcinoma specimen (n = 191). Moreover, RANKL expression was measured in cancer cell lines (BICR3, BICR56) by immunohistochemistry and western blot analysis. Scanned images were digitally analyzed using ImageJ and the immunomembrane plug-in. CD44 and RANKL expression on protein level was correlated with clinical characteristics and impact on survival. RANKL was co-labeled with CD44 in immunohistochemical and immunofluorescence double labeling experiments. Although high CD44+/RANKL+ co-expression was significantly associated with clinicopathological factors and worse survival, multivariate analysis did not demonstrate high CD44+/RANKL+ co-expression as independent prognostic factor. Immunohistochemical and immunofluorescence double labeling experiments revealed RANKL expression by CD44+ cancer cells. RANKL specificity was confirmed by western blot analysis. For the first time, this study provides evidence that RANKL expression in OSCC might be associated with disease recurrence and a cell compartment measured by CD44+/RANKL+ co-expression within the mucosal epithelial basal layer cells.
核因子κB受体激活剂（RANK）/核因子κB受体激活剂配体（RANKL）信号通路有助于假定的癌症干细胞（CSC）维持其干性。分析了191例口腔鳞状细胞癌标本中CD44和RANKL的表达。此外，通过免疫组织化学和蛋白质印迹分析测定了癌细胞系（BICR3、BICR56）中的RANKL表达。使用ImageJ和免疫膜插件对扫描图像进行数字分析。蛋白质水平上的CD44和RANKL表达与临床特征及对生存的影响相关。在免疫组织化学和免疫荧光双重标记实验中，RANKL与CD44共同标记。尽管高CD44+/RANKL+共表达与临床病理因素及较差的生存率显著相关，但多因素分析未显示高CD44+/RANKL+共表达为独立的预后因素。免疫组织化学和免疫荧光双重标记实验显示CD44+癌细胞表达RANKL。蛋白质印迹分析证实了RANKL的特异性。本研究首次提供证据表明，口腔鳞状细胞癌中RANKL的表达可能与疾病复发以及黏膜上皮基底层细胞中通过CD44+/RANKL+共表达测量的细胞区室有关。
