Suppression of keratinocyte stratification by a dominant negative JunB mutant without blocking cell proliferation
Suppression of keratinocyte stratification by a dominant negative JunB mutant without blocking cell proliferation
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DOI：
10.1111/j.1365-2443.2007.01043.x
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发表时间：
2007-02-01
期刊：
影响因子：
2.1
通讯作者：
Kato, Mitsuyasu
中科院分区：
文献类型：
--
作者：
Ikebe, Dai;Wang, Bei;Kato, Mitsuyasu
角质形成细胞在气液界面培养时形成分层的表皮样结构。角质形成细胞的三维（3D）培养已成功使用超过25年，但仍不清楚角质形成细胞分层的原因。AP-1蛋白是一种普遍存在的转录因子，调节许多生物学过程，包括细胞增殖、分化和凋亡。我们建立了HaCaT-JunB Delta N，这是一种表达突变型JunB的人角质形成细胞系，该突变型JunB对AP-1活性具有显性负效应。HaCaT-JunB Delta N细胞的分层在3D培养条件下明显受到抑制，其中HaCaT细胞分层类似于分层鳞状上皮。然而，HaCaT-JunB Delta N细胞在二维（2D）和3D培养条件下的增殖活性与HaCaT细胞的增殖活性非常相似。为了筛选负责不同分层能力的候选基因，我们检测了空气暴露前后HaCaT细胞的基因表达谱。几个具有抗氧化功能的基因，如醛酮还原酶和硒蛋白P在HaCaT细胞中的空气暴露后高度表达，但在HaCaT-JunB Delta N细胞中没有表达。我们的研究结果表明，当HaCaT细胞在3D培养条件下形成分层表皮样结构时，AP-1活性存在一种新的作用。
Keratinocytes make a stratified epidermoid structure when cultured at an air-liquid interface. The three-dimensional (3D) culture of keratinocytes has been successfully used for more than 25 years, but it is still unclear why keratinocytes stratify in response to air exposure. AP-1 proteins are ubiquitous transcription factors that regulate many biological processes, including cell proliferation, differentiation and apoptosis. We established HaCaT-JunB Delta N, a human keratinocyte cell line that expressed a mutant JunB with a dominant negative effect on AP-1 activity. Stratification of HaCaT-JunB Delta N cells was markedly suppressed in a 3D culture condition, in which HaCaT cells stratified similarly to stratified squamous epithelia. However, HaCaT-JunB Delta N cells had proliferation activities that were closely equivalent to those of HaCaT cells, under both two-dimensional (2D) and 3D culture conditions. To screen for the candidate gene responsible for the different stratification ability, we examined the gene expression profile of HaCaT cells before and after air exposure. Several genes with an antioxidative function, such as aldo-keto reductase and selenoprotein P were highly expressed after air exposure in HaCaT cells but not in HaCaT-JunB Delta N cells. Our findings indicate the presence of a novel role of AP-1 activity when HaCaT cells make a stratified epidermoid structure under 3D culture conditions.