Division of Oral Biotechnology, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Hugstetter Str. 55, 79106 Freiburg, Germany; Faculty of Engineering, University of Freiburg, Georges-Köhler-Allee 101, 79110 Freiburg, Germany.
Department of Orthodontics, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, Hugstetter Str. 55, 79106 Freiburg, Germany; Faculty of Biology, University of Freiburg, Schaenzlestr. 1, 79104 Freiburg, Germany.
Biochim Biophys Acta Mol Cell Res. 2018 Jan;1865(1):209-219. doi: 10.1016/j.bbamcr.2017.10.015. Epub 2017 Nov 2.
Accumulating evidences indicate that alcohol might play a causative in oral cancer. Unfortunately, in vitro cell systems, uncovering the molecular background of the underlying cell transformation process, are rare. Therefore, this study was conducted, to identify molecular changes and characterize their putative cell behavioral consequences in epitheloid (EPI) and fibroblastoid (FIB) oral keratinocyte phenotypes, arising from chronical alcohol treatment. Concerning adherens junctions (AJs), both EPI and FIB showed membrane-bound β-catenin, but exhibited differences for E-cadherin and zyxin. While EPI revealed E-cadherin/β-catenin membrane co-localization, which in parts also applied for zyxin, FIB membranes were devoid of E-cadherin and exhibited marginal zyxin expression. Fetal calf serum (FCS) administration in starved cells promoted proliferation in both keratinocyte phenotypes, whereat EPI and FIB yielded a strikingly modified FCS sensitivity on the temporal scale. Impedance measurement-based cell index detection yielded proliferation stimulation occurring much earlier in FIB (<20h) compared to EPI (>45h). Nuclear preference of the proliferation-associated YAP co-transcription factor in FIB was FCS independent, while it required FCS in EPI. Taken together, the lack of membrane-inherent E-cadherin/β-catenin co-localization together with low zyxin - reveals perturbation of AJ integrity in FIB. Regarding cell behavior, perturbed AJs in FIB correlate with temporal proliferation sensitivity towards FCS. CYF of 5.6 strongly suggests involvement of chromatin-bound YAP in FIB's proliferation temperosensitivity. These molecular differences detected for EPI and FIB are part of the underlying cell transformation process of alcohol-induced oral carcinogenesis, and indicate FIB being in a more advanced transformation stage.
越来越多的证据表明，酒精可能是口腔癌的致病因素。不幸的是，很少有体外细胞系统能够揭示潜在的细胞转化过程的分子背景。因此，本研究旨在鉴定分子变化，并在慢性酒精处理引起的上皮（EPI）和纤维母细胞样（FIB）口腔角质形成细胞表型中描述其潜在的细胞行为后果。关于黏附连接（AJs），EPI 和 FIB 均显示膜结合的β-连环蛋白，但 E-钙黏蛋白和 zyxin 存在差异。虽然 EPI 显示 E-钙黏蛋白/β-连环蛋白膜共定位，部分也适用于 zyxin，但 FIB 膜缺乏 E-钙黏蛋白，并且表达边缘化的 zyxin。在饥饿细胞中添加胎牛血清（FCS）可促进两种角质形成细胞表型的增殖，其中 EPI 和 FIB 在时间尺度上表现出明显不同的 FCS 敏感性。基于阻抗测量的细胞指数检测表明，FIB 中的增殖刺激发生得更早（<20h），而 EPI 则发生得更晚（>45h）。FIB 中与增殖相关的 YAP 共转录因子的核偏好与 FCS 无关，而在 EPI 中则需要 FCS。总之，FIB 中膜固有 E-钙黏蛋白/β-连环蛋白共定位的缺乏以及 zyxin 表达水平低表明 AJ 完整性受到干扰。就细胞行为而言，FIB 中 AJ 的紊乱与 FCS 对增殖的时间敏感性相关。CYF 为 5.6 强烈表明，染色质结合的 YAP 参与了 FIB 的增殖温度敏感性。EPI 和 FIB 检测到的这些分子差异是酒精诱导口腔癌变中细胞转化过程的一部分，表明 FIB 处于更高级的转化阶段。
