Laboratory of Biochemistry, Scientific Affairs, APIVITA SA, Industrial Park of Markopoulo Mesogaias, 19003, Markopoulo Attikis, Athens, Greece.
Comparative Endocrinology and Integrative Biology Group (CEIB), Centro de Ciências Do Mar (CCMAR), Universidade Do Algarve, Campus de Gambelas, 8005-139, Faro, Portugal.
Biochimie. 2020 Apr-May;171-172:72-78. doi: 10.1016/j.biochi.2020.02.008. Epub 2020 Feb 18.
Cartilage acidic protein 1 (CRTAC1) is an extracellular matrix protein of human chondrogenic tissue that is also present in other vertebrates, non-vertebrate eukaryotes and in some prokaryotes. The function of CRTAC1 remains unknown but the protein's structure indicates a role in cell-cell or cell-matrix interactions and calcium-binding. The aim of the present study was to evaluate the in vitro effects of hCRTAC1-A on normal human dermal fibroblasts (NHDF). A battery of in vitro assays (biochemical and PCR), immunofluorescence and a biosensor approach were used to characterize the protein's biological activities on NHDF cells in a scratch assay. Gene expression analysis revealed that hCRTAC1-A protein is associated with altered levels of expression for genes involved in the processes of cell proliferation (CXCL12 and NOS2), cell migration (AQP3 and TNC), and extracellular matrix-ECM regeneration and remodeling (FMOD, TIMP1, FN1) indicating a role for hCRTAC1-A in promoting these activities in a scratch assay. In parallel, the candidate processes identified by differential gene transcription were substantiated and extended using Electric cell-substrate impedance sensing (ECIS) technology, immunofluorescence and cell viability assays. Our findings indicate that hCRTAC1-A stimulated cell proliferation, migration and ECM production in primary human fibroblasts in vitro.
软骨酸性蛋白 1(CRTAC1)是人类软骨组织的细胞外基质蛋白,也存在于其他脊椎动物、非脊椎动物真核生物和一些原核生物中。CRTAC1 的功能尚不清楚,但该蛋白的结构表明其在细胞-细胞或细胞-基质相互作用和钙结合中起作用。本研究旨在评估 hCRTAC1-A 对正常人真皮成纤维细胞(NHDF)的体外影响。使用一系列体外测定(生化和 PCR)、免疫荧光和生物传感器方法,在划痕实验中对 NHDF 细胞中蛋白的生物学活性进行了表征。基因表达分析表明,hCRTAC1-A 蛋白与参与细胞增殖(CXCL12 和 NOS2)、细胞迁移(AQP3 和 TNC)以及细胞外基质-ECM 再生和重塑(FMOD、TIMP1、FN1)过程的基因表达水平改变相关,这表明 hCRTAC1-A 在促进划痕实验中的这些活性中起作用。平行地,使用电动细胞-底物阻抗感应(ECIS)技术、免疫荧光和细胞活力测定,对差异基因转录鉴定的候选过程进行了证实和扩展。我们的研究结果表明,hCRTAC1-A 在体外刺激原代人成纤维细胞的增殖、迁移和 ECM 产生。