Expression of wild-type, but not mutant, loricrin causes programmed cell death in HaCaT keratinocytes
Expression of wild-type, but not mutant, loricrin causes programmed cell death in HaCaT keratinocytes
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野生型（而非突变型）兜甲素的表达导致 HaCaT 角质形成细胞程序性细胞死亡
DOI：
10.1111/j.1346-8138.2010.00932.x
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发表时间：
2010
期刊：
影响因子：
3.1
通讯作者：
Kubota Y
中科院分区：
文献类型：
--
作者：
Yoneda K;Demitsu T;Manabe M;Igarashi J;Kosaka H;Inagaki N;Takahashi H;Kon A;Kakurai M;Kubota Y
表皮角化细胞包膜是一种复杂的蛋白质-脂质复合物，它取代角质细胞的质膜，对表皮屏障功能至关重要。 Loricrin 是表皮角化细胞包膜的主要成分，约占质量的 70%。为了探索除表皮角化细胞包膜主要成分之外的野生型（WT）兜甲蛋白的新功能，我们在HaCaT角质形成细胞中瞬时表达编码人WT和突变体兜甲蛋白（730insG）的构建体。转染WT或突变型loricrin的HaCaT细胞处于分化水平。转染细胞中WT兜甲蛋白弥漫于细胞质和细胞核中。在 WT loricrin 转染的 HaCaT 角质形成细胞的细胞核中观察到阳性转移酶脱氧苷基尿苷末端标记染色。 DNA 片段化测定的数据表明，与突变型兜甲蛋白相比，仅野生型兜甲蛋白诱导 DNA 梯子。 WT 兜甲蛋白转染的 HaCaT 角质形成细胞对程序性细胞死亡 (PCD) 敏感。还观察到 caspase-14 的激活。相比之下，突变型兜甲素转染的 HaCaT 细胞中没有发生 PCD 或 caspase-14 的激活。这些结果表明，WT loricrin 的表达促进 HaCaT 角质形成细胞中 PCD 的诱导。
The epidermal cornified cell envelope is a complex protein-lipid composite that replaces the plasma membrane of corneocytes and is crucial for epidermal barrier function. Loricrin is a major constituent of the epidermal cornified cell envelope, contributing approximately 70% by mass. In order to explore novel function of wild-type (WT) loricrin other than the major component of the epidermal cornified cell envelope, we transiently expressed construct encoding human WT and mutant loricrin (730insG) in HaCaT keratinocytes. HaCaT cells transfected with WT or mutant loricrin were at differentiation level. WT loricrin in the transfected cells was seen diffusely in the cytoplasm and nuclei. Positive transferase deoxytidyl uridine end labeling staining was observed in the nuclei of WT loricrin-transfected HaCaT keratinocytes. Data from the DNA fragmentation assay showed that only WT loricrin induced DNA ladders compared with that of mutant loricrin. WT loricrin-transfected HaCaT keratinocytes were susceptible to programmed cell death (PCD). Activation of caspase-14 was also seen. In contrast, PCD or activation of caspase-14 did not occur in mutant loricrin-transfected HaCaT cells. These results suggest that the expression of WT loricrin facilitates induction of PCD in HaCaT keratinocytes.