{"title":"ATP2C1 基因敲除诱导细胞骨架和紧密连接蛋白的异常表达,模拟海利-海利病","authors":"Mingzhu Zhou, Shiran Kang, Yumin Xia, Dingwei Zhang, Wenwen Chen","doi":"10.25259/ijdvl_853_2023","DOIUrl":null,"url":null,"abstract":"\n\nHailey–Hailey disease (HHD) is a rare, autosomal dominant, hereditary skin disorder characterised by epidermal acantholysis. The HHD-associated gene ATPase calcium-transporting type 2C member 1 (ATP2C1) encodes the protein secretory pathway Ca2+ ATPase1 (SPCA1), playing a critical role in HHD pathogenesis.\n\n\n\nWe aimed to investigate the effect of ATP2C1 knockdown on keratinocytes that mimicked acantholysis in HHD. \n\n\n\nImmunohistochemistry (IHC) was employed to evaluate the levels of cytoskeletal and tight junction proteins such as SPCA1, P-cofilin, F-actin, claudins, occludin, and zonula occludens 1 in the skin biopsies of patients with HHD. Subsequently, the expression of these proteins in cultured ATP2C1 knockdown keratinocytes was analysed using Western blotting and immunofluorescence. Furthermore, we assessed the proliferation, apoptosis, and intracellular Ca2+ concentrations in the ATP2C1-knocked keratinocytes.\n\n\n\nThe results showed decreased levels of these proteins (SPCA1, P-cofilin, F-actin, claudins, occluding, and zonula occludens 1) in HHD skin lesions. Moreover, their levels decreased in human keratinocytes transfected with ATP2C1 short hairpin RNA, accompanied by morphological acantholysis. Furthermore, the proliferation and apoptosis of the keratinocytes, as well as intracellular calcium concentrations in these cells, were not affected.\n\n\n\nThe limitations of this study are the absence of animal experiments and the failure to explore the relationship between skeletal and tight junction proteins.\n\n\n\nThe present study indicated that ATP2C1 inhibition led to abnormal levels of the cytoskeletal and tight junction proteins in the keratinocytes. Therefore, keratinocytes can mimic HHD-like acantholysis and serve as an in vitro model, helping develop treatment strategies against HHD.\n","PeriodicalId":513160,"journal":{"name":"Indian Journal of Dermatology, Venereology and Leprology","volume":"44 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2024-05-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"ATP2C1 knockdown induces abnormal expressions of cytoskeletal and tight junction proteins mimicking Hailey–Hailey disease\",\"authors\":\"Mingzhu Zhou, Shiran Kang, Yumin Xia, Dingwei Zhang, Wenwen Chen\",\"doi\":\"10.25259/ijdvl_853_2023\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"\\n\\nHailey–Hailey disease (HHD) is a rare, autosomal dominant, hereditary skin disorder characterised by epidermal acantholysis. The HHD-associated gene ATPase calcium-transporting type 2C member 1 (ATP2C1) encodes the protein secretory pathway Ca2+ ATPase1 (SPCA1), playing a critical role in HHD pathogenesis.\\n\\n\\n\\nWe aimed to investigate the effect of ATP2C1 knockdown on keratinocytes that mimicked acantholysis in HHD. \\n\\n\\n\\nImmunohistochemistry (IHC) was employed to evaluate the levels of cytoskeletal and tight junction proteins such as SPCA1, P-cofilin, F-actin, claudins, occludin, and zonula occludens 1 in the skin biopsies of patients with HHD. Subsequently, the expression of these proteins in cultured ATP2C1 knockdown keratinocytes was analysed using Western blotting and immunofluorescence. Furthermore, we assessed the proliferation, apoptosis, and intracellular Ca2+ concentrations in the ATP2C1-knocked keratinocytes.\\n\\n\\n\\nThe results showed decreased levels of these proteins (SPCA1, P-cofilin, F-actin, claudins, occluding, and zonula occludens 1) in HHD skin lesions. Moreover, their levels decreased in human keratinocytes transfected with ATP2C1 short hairpin RNA, accompanied by morphological acantholysis. Furthermore, the proliferation and apoptosis of the keratinocytes, as well as intracellular calcium concentrations in these cells, were not affected.\\n\\n\\n\\nThe limitations of this study are the absence of animal experiments and the failure to explore the relationship between skeletal and tight junction proteins.\\n\\n\\n\\nThe present study indicated that ATP2C1 inhibition led to abnormal levels of the cytoskeletal and tight junction proteins in the keratinocytes. Therefore, keratinocytes can mimic HHD-like acantholysis and serve as an in vitro model, helping develop treatment strategies against HHD.\\n\",\"PeriodicalId\":513160,\"journal\":{\"name\":\"Indian Journal of Dermatology, Venereology and Leprology\",\"volume\":\"44 1\",\"pages\":\"\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2024-05-04\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Indian Journal of Dermatology, Venereology and Leprology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.25259/ijdvl_853_2023\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Indian Journal of Dermatology, Venereology and Leprology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.25259/ijdvl_853_2023","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
摘要
海利-海利病(Hailey-Hailey disease,HHD)是一种罕见的常染色体显性遗传性皮肤病,以表皮棘层溶解为特征。HHD相关基因ATPase calcium-transporting type 2C member 1(ATP2C1)编码蛋白分泌途径Ca2+ ATPase1(SPCA1),在HHD发病机制中起着关键作用。我们采用免疫组织化学(IHC)方法评估了HHD患者皮肤活检组织中细胞骨架和紧密连接蛋白的水平,如SPCA1、P-cofilin、F-actin、claudins、occludin和zonula occludens 1。随后,我们使用 Western 印迹法和免疫荧光法分析了这些蛋白在培养的 ATP2C1 敲除角质形成细胞中的表达情况。结果显示,这些蛋白质(SPCA1、P-cofilin、F-actin、claudins、occluding 和 zonula occludens 1)的水平在 HHD 皮肤病变中有所下降。此外,在转染了 ATP2C1 短发夹核糖核酸的人类角质细胞中,这些蛋白的水平也有所下降,并伴有形态学上的棘层溶解。本研究的局限性在于没有进行动物实验,也没有探讨骨骼蛋白和紧密连接蛋白之间的关系。因此,角朊细胞可以模拟HHD样棘层溶解,并作为体外模型,帮助开发针对HHD的治疗策略。
ATP2C1 knockdown induces abnormal expressions of cytoskeletal and tight junction proteins mimicking Hailey–Hailey disease
Hailey–Hailey disease (HHD) is a rare, autosomal dominant, hereditary skin disorder characterised by epidermal acantholysis. The HHD-associated gene ATPase calcium-transporting type 2C member 1 (ATP2C1) encodes the protein secretory pathway Ca2+ ATPase1 (SPCA1), playing a critical role in HHD pathogenesis.
We aimed to investigate the effect of ATP2C1 knockdown on keratinocytes that mimicked acantholysis in HHD.
Immunohistochemistry (IHC) was employed to evaluate the levels of cytoskeletal and tight junction proteins such as SPCA1, P-cofilin, F-actin, claudins, occludin, and zonula occludens 1 in the skin biopsies of patients with HHD. Subsequently, the expression of these proteins in cultured ATP2C1 knockdown keratinocytes was analysed using Western blotting and immunofluorescence. Furthermore, we assessed the proliferation, apoptosis, and intracellular Ca2+ concentrations in the ATP2C1-knocked keratinocytes.
The results showed decreased levels of these proteins (SPCA1, P-cofilin, F-actin, claudins, occluding, and zonula occludens 1) in HHD skin lesions. Moreover, their levels decreased in human keratinocytes transfected with ATP2C1 short hairpin RNA, accompanied by morphological acantholysis. Furthermore, the proliferation and apoptosis of the keratinocytes, as well as intracellular calcium concentrations in these cells, were not affected.
The limitations of this study are the absence of animal experiments and the failure to explore the relationship between skeletal and tight junction proteins.
The present study indicated that ATP2C1 inhibition led to abnormal levels of the cytoskeletal and tight junction proteins in the keratinocytes. Therefore, keratinocytes can mimic HHD-like acantholysis and serve as an in vitro model, helping develop treatment strategies against HHD.