Discrimination of epithelium-like and fibroblast-like phenotypes derived from ethanol-treated immortalised human gingival keratinocytes in epithelial equivalents
Discrimination of epithelium-like and fibroblast-like phenotypes derived from ethanol-treated immortalised human gingival keratinocytes in epithelial equivalents
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上皮等同物中乙醇处理的永生化人牙龈角质形成细胞衍生的上皮样和成纤维细胞样表型的区分
DOI：
10.1007/s00441-007-0551-y
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发表时间：
2008
影响因子：
3.6
通讯作者：
P. Tomakidi
中科院分区：
文献类型：
--
作者：
E. Müssig;T. Steinberg;A. Kohl;W. Chamulitrat;G. Komposch;P. Tomakidi
乙醇处理永生化人牙龈角化细胞（IHGK）产生上皮样（EPI）和成纤维细胞样（FIB）表型。关于分层的牙龈上皮，迄今为止，尚未在三维组织/上皮环境中分析将细胞分配给这些表型的推定结构和分子差异。因此，我们在IHGK、EPI和FIB细胞共培养1周和2周后生成上皮等效物（epithelial equivalents, EEs），并对上皮生物标志物（包括角蛋白K14、整合素亚基α6和β1、E-cadherin和间充质vimentin）的EEs进行蛋白和基因表达研究。与IHGK和EPI的EEs一样，间接免疫荧光显示FIB细胞EEs中β1整合素持续表达。然而，FIB细胞在第2周表现出K14和整合素α6蛋白的显著下调和E-cadherin的缺失，而vimentin升高。尽管其他基因的转录在所有表型中保持不变，但FIB EEs在两个时间点都缺乏e -钙粘蛋白的转录本。因此，FIB表型表现出较差的上皮结构，与上皮生物标志物的表达紊乱和间充质波形蛋白的持久性一致。转录分析显示FIB细胞中IHGK和EPI以及K14和α6的vimentin转录后调控。我们的研究结果表明，EEs中上皮完整性和分子表达的差异允许EPI和FIB细胞的区分。这表明FIB细胞具有上皮-间质转化的特征，反映了上皮细胞转化的一个更先进的阶段。
Ethanol treatment of immortalised human gingival keratinocytes (IHGK) yields in an epithelium-like (EPI) and fibroblast-like (FIB) phenotype. With respect to the stratified gingival epithelium, putative structural and molecular differences assigning cells to these phenotypes have not, to date, been analysed in a three-dimensional tissue/epithelial context. Therefore, we generated epithelial equivalents (EEs) in organotypic co-cultures of IHGK, EPI and FIB cells for 1 and 2 weeks and conducted protein and gene expression studies on the EEs for epithelial biomarkers including keratin K14, integrin subunits α6 and β1, E-cadherin, and mesenchymal vimentin. As in the EEs of IHGK and EPI, indirect immunofluorescence revealed continuous expression of β1 integrin in EEs of FIB cells. However, FIB cells exhibited a significant down-regulation in K14 and integrin α6 protein and a loss of E-cadherin at week 2, whereas vimentin was increased. FIB EEs were devoid of transcripts for E-cadherin at both time points, although transcription of the other genes remained constant in all phenotypes. Thus, the FIB phenotype exhibited a poor epithelial structure coinciding with disturbances in the expression of epithelial biomarkers and the persistence of mesenchymal vimentin. Transcription analysis revealed post-transcriptional regulation of vimentin in IHGK and EPI and of K14 and α6 in FIB cells. Our findings indicate that differences in the epithelial integrity and expression of molecules in EEs allow for the discrimination of EPI and FIB cells. This suggests that FIB cells share features of epithelial-mesenchymal transition and reflect a more progressive stage in epithelial cell transformation.
DOI：
--
发表时间：
2001
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
N. Bhowmick;R. Zent;M. Ghiassi;M. McDonnell;H. Moses
通讯作者：
N. Bhowmick;R. Zent;M. Ghiassi;M. McDonnell;H. Moses