细胞外表吗啡会影响 HaCaT 角质细胞中 profilaggrin 的表达和处理。

IF 2 4区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Cytotechnology Pub Date : 2023-04-01 Epub Date: 2022-12-15 DOI:10.1007/s10616-022-00566-8
Haruna Hori, Ayaka Kotani, Junya Abe, Shuji Matsuguchi, Yohei Hirai
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引用次数: 0

摘要

丝胶蛋白是皮肤表皮中的一种丝状相关蛋白,它的表达和加工与角质细胞的粟粒化密切相关。大型前体丝胶蛋白(Pro-FLG)最初在颗粒层的角质透明质颗粒中被检测到,随后被加工成 10 到 12 个丝胶蛋白单体(mFLGs)用于角质组装,最终降解成较小的肽,在表皮最外层作为天然保湿因子(NMF)发挥作用。我们以前曾报道过,在外部刺激下挤出的表吗啡(EPM)会严重干扰表皮的末端分化。我们利用具有诱导表达和重组 EPM 及 FLG 的 HaCaT 角质细胞,研究了细胞外 EPM 对丝胶蛋白表达谱的影响。由于原代角朊细胞中 Pro-FLG 的表达和处理伴随着细胞凋亡,我们采用了在标准培养基中生长和表达丝胶蛋白 mRNA 的 HaCaT 角朊细胞。在细胞外 EPM 的异位刺激下,Pro-FLG 的表达随着细胞中角质透明蛋白颗粒的消失而减少,而 filaggrin mRNA 保持不变,profilaggrin 的加工也没有加速。此外,我们使用一种重组形式的 mFLG 进行细胞内定位,发现细胞外的 EPM 会阻碍 mFLG 蛋白质分解以产生 NMF。综上所述,细胞外挤出的EPM是一种表皮粟粒化阻滞剂,它不仅能降低Pro-FLG的表达,还能减少HaCaT角质细胞中NMF的产生:在线版本包含补充材料,可查阅 10.1007/s10616-022-00566-8。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Extracellular epimorphin impairs expression and processing of profilaggrin in HaCaT keratinocytes.

Extracellular epimorphin impairs expression and processing of profilaggrin in HaCaT keratinocytes.

The expression and processing of filaggrin, a filament-associated protein in the skin epidermis, is closely associated with keratinocyte cornification. The large precursor profilaggrin (Pro-FLG) is initially detected at the granular layer in keratohyalin granules, subsequently processed into 10 to 12 filaggrin monomers (mFLGs) for keratin assembly, and ultimately degraded into smaller peptides that behave as natural moisturizing factor (NMF) at the outermost epidermis. We previously reported that epimorphin (EPM) extruded upon external stimuli severely perturbs epidermal terminal differentiation. Using HaCaT keratinocytes with inducible expression and recombinant EPM and FLG, we investigated the effect of extracellular EPM on the expression profile of filaggrin. As expression and processing of Pro-FLG in primary keratinocytes are accompanied with apoptotic cell death, we employed HaCaT keratinocytes that grow and express filaggrin mRNA in standard culture medium. In response to ectopic stimulation with extracellular EPM, Pro-FLG expression decreased with elimination of keratohyalin granules in the cells, with filaggrin mRNA remained constant and profilaggrin processing was not accelerated. Additionally, using a recombinant form of mFLG engineered for intracellular localization, we found that extracellular EPM hindered proteolytic cleavage of mFLG for production of NMF. Taken together, extracellularly extruded EPM, an epidermal cornification blocker, not only decreases Pro-FLG expression but also reduces the production of NMF in HaCaT keratinocytes.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-022-00566-8.

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来源期刊
Cytotechnology
Cytotechnology 生物-生物工程与应用微生物
CiteScore
4.10
自引率
0.00%
发文量
49
审稿时长
6-12 weeks
期刊介绍: The scope of the Journal includes: 1. The derivation, genetic modification and characterization of cell lines, genetic and phenotypic regulation, control of cellular metabolism, cell physiology and biochemistry related to cell function, performance and expression of cell products. 2. Cell culture techniques, substrates, environmental requirements and optimization, cloning, hybridization and molecular biology, including genomic and proteomic tools. 3. Cell culture systems, processes, reactors, scale-up, and industrial production. Descriptions of the design or construction of equipment, media or quality control procedures, that are ancillary to cellular research. 4. The application of animal/human cells in research in the field of stem cell research including maintenance of stemness, differentiation, genetics, and senescence, cancer research, research in immunology, as well as applications in tissue engineering and gene therapy. 5. The use of cell cultures as a substrate for bioassays, biomedical applications and in particular as a replacement for animal models.
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