Comparison of gene expression profiles in human keratinocyte mono-layer cultures, reconstituted epidermis and normal human skin; transcriptional effects of retinoid treatments in reconstituted human epidermis
Comparison of gene expression profiles in human keratinocyte mono-layer cultures, reconstituted epidermis and normal human skin; transcriptional effects of retinoid treatments in reconstituted human epidermis
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DOI：
10.1034/j.1600-0625.2002.110107.x
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发表时间：
2002-02-01
影响因子：
3.6
通讯作者：
Deguercy, A
中科院分区：
文献类型：
--
作者：
Bernard, FX;Pedretti, N;Deguercy, A
为了验证用于皮肤病药物预测筛选的模型，我们使用定制的包含475个皮肤相关基因的cDNA宏阵列系统来分析来自不同来源的人角质形成细胞的基因表达模式：(1)正常人表皮角质形成细胞单层培养，(2)商品化的Skinethic重组人表皮模型，(3)正常人表皮的活检组织。那些在角质形成细胞单层或重组表皮中显着检测到的标记物，在正常表皮活检组织中很少未检测到或检测到极低水平。100多个标记物在正常表皮和重组表皮中均有表达，但仅有90%在角质形成细胞单层中表达；几种终末分化标记物，如微丝蛋白、氯霉素和新冠桥连蛋白在正常表皮和重组表皮中表达较强，但在角质形成细胞单层中表达不明显。在本文描述的实验条件下，重组的人类表皮模型被发现显著地再现了正常人类表皮的基因表达谱。然后，使用相同的方法，我们研究了全反式维甲酸、9-顺式维甲酸、全反式视黄醇和商业化的维甲酸乳膏(Retacnyl(TM))对重组人表皮基因表达谱的影响。根据处理的性质和长度，发现有40多个基因被显著修改。在表达下调的基因中，我们发现了细胞角蛋白1、10、2E和6B，几个角化的包膜前体，整合素α3、α6、β1、β4，桥粒、半桥粒和表皮基底膜的一些成分。角蛋白18和19，自分泌和旁分泌生长因子，如HB-EGF，IGF-1，PDGFA，钙粒蛋白A和B，IL-1α和其他IL-1相关标志物，II型IL-1受体和I型IL-1受体拮抗剂的转录上调。我们的结果证实了大多数已知的维甲酸对人类表皮的影响，但也为了解它们在皮肤上的复杂药理活性提供了新的见解。使用的重组人表皮被证明是一个高度预测的模型，用于评估皮肤靶向化合物的疗效，如维甲酸。
In order to validate a model for predictive screening of dermatological drugs, we used a customized cDNA macro-array system containing 475 skin-related genes to analyze the gene expression patterns in human keratinocytes from different origins: (1) normal human epidermal keratinocyte mono-layer cultures, (2) the commercially available SkinEthic reconstituted human epidermis model, and (3) biopsies of normal human epidermis. Few markers of those that were detected significantly in keratinocyte mono-layers or in reconstituted epidermis were undetected or detected at very low level in the normal epidermis biopsies. A comparative expression of more than 100 markers could be evidenced in both normal epidermis and reconstituted epidermis samples; however, only 90% of these were detected in keratinocyte monolayers: expression of several terminal differentiation markers, such as filaggrin, loricrin, and corneodesmosin were strongly detected in normal epidermis and reconstituted epidermis, but were not significantly expressed in keratinocyte mono-layers. Under the experimental conditions described herein, the reconstituted human epidermis model was found to significantly reproduce the gene expression profile of normal human epidermis. Using the same methodology, we then investigated the effects of all-trans retinoic acid, 9-cis retinoic acid, all-trans retinol and a commercialized tretinoin-containing cream (Retacnyl(TM)) on the gene expression profiles of reconstituted human epidermis. According to the nature and the length of the treatments, more than 40 genes were found significantly modified. Among the genes whose expression was decreased, we found cytokeratins 1, 10, 2E, and 6B, several cornified envelope precursors, integrins alpha3, alpha6, beta1, beta4, some components of desmosomes, of hemi-desmosomes and of the epidermal basement membrane. Transcriptional upregulation was observed for keratins 18 and 19, autocrine and paracrine growth factors such as HB-EGF, IGF 1, PDGFA, calgranulins A and B, interleukin-1alpha and the other IL-1-related markers, type II IL-1 receptor and type I IL-1-receptor antagonist. Our results confirm most of the known effects of retinoids on human epidermis, but also give new insights into their complex pharmacological activity on skin. The reconstituted human epidermis used proves to be a highly predictive model for efficacy evaluation of skin-targeted compounds, such as retinoids.