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Augmented osteogenesis of mesenchymal stem cells using a fragmented Runx2 mixed with cell-penetrating, dimeric a-helical peptide.
European Journal of Pharmaceutical Sciences ( IF 4.3 ) Pub Date : 2019-12-30 , DOI: 10.1016/j.ejps.2019.105210 So Hee Nam 1 , Yan Lee 1 , Joon Hyung Ahn 1 , Chun Kee Chung 2 , Hee-Jin Yang 3 , Sung Bae Park 3 , Sangmok Jang 4
European Journal of Pharmaceutical Sciences ( IF 4.3 ) Pub Date : 2019-12-30 , DOI: 10.1016/j.ejps.2019.105210 So Hee Nam 1 , Yan Lee 1 , Joon Hyung Ahn 1 , Chun Kee Chung 2 , Hee-Jin Yang 3 , Sung Bae Park 3 , Sangmok Jang 4
Affiliation
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The intracellular delivery of transcription factor/cofactor using cell penetrating peptide (CPP) can lead to selective osteogenesis. The present work investigates the cell-penetrating potential of the a cyclic, α-helical cell-penetrating peptide based on leucine and lysine residues (cLK) for intracellular delivery in MC3T3 cells and the osteogenic effects of a C-terminal proline‑serine‑threonine-rich (PST) domain of Runx2 using cLK in rat mesenchymal stem cells (MSCs). We confirmed that the combination of cLK and fluorescein 5-isothiocyanate (FITC)-fragmented-Runx2 (fRunx2) showed an enhanced cell-penetrating activity of FITC-fRunx2 compared with FITC-fRunx2 alone. In addition, the fRunx2-cLK group showed strong staining with alizarin red compared with other groups and the degree of alizarin red staining in the fRunx2-cLK group was also 1.2-fold higher than that in the fRunx2-Tat group. The ALP and osteocalcin gene expression levels in the fRunx2-cLK group were higher than those in the other groups. The fRunx2 transferred effectively into the cytoplasm aided by the cLK peptide and augmented the osteogenic differentiation of MSCs.
中文翻译：
使用片段Runx2与穿透细胞的二聚a-螺旋肽混合，增强间充质干细胞的成骨作用。
使用细胞穿透肽（CPP）在细胞内传递转录因子/辅因子可以导致选择性成骨。本工作研究了基于亮氨酸和赖氨酸残基（cLK）的环状α螺旋细胞穿透肽在MC3T3细胞中向细胞内传递的细胞穿透潜力以及C末端脯氨酸-丝氨酸-苏氨酸的成骨作用大鼠间充质干细胞（MSCs）中使用cLK实现Runx2的富含（PST）域。我们确认，与单独的FITC-fRunx2相比，cLK和5异硫氰酸荧光素（FITC）片段化Runx2（fRunx2）的组合显示出增强的FITC-fRunx2细胞穿透活性。另外，与其他组相比，fRunx2-cLK组的茜素红染色强烈，fRunx2-cLK组的茜素红染色程度也为1。比fRunx2-Tat组高2倍。fRunx2-cLK组的ALP和骨钙素基因表达水平高于其他组。fRunx2在cLK肽的辅助下有效转移到细胞质中，并增强了MSC的成骨分化。
更新日期：2019-12-31
中文翻译：
使用片段Runx2与穿透细胞的二聚a-螺旋肽混合，增强间充质干细胞的成骨作用。
使用细胞穿透肽（CPP）在细胞内传递转录因子/辅因子可以导致选择性成骨。本工作研究了基于亮氨酸和赖氨酸残基（cLK）的环状α螺旋细胞穿透肽在MC3T3细胞中向细胞内传递的细胞穿透潜力以及C末端脯氨酸-丝氨酸-苏氨酸的成骨作用大鼠间充质干细胞（MSCs）中使用cLK实现Runx2的富含（PST）域。我们确认，与单独的FITC-fRunx2相比，cLK和5异硫氰酸荧光素（FITC）片段化Runx2（fRunx2）的组合显示出增强的FITC-fRunx2细胞穿透活性。另外，与其他组相比，fRunx2-cLK组的茜素红染色强烈，fRunx2-cLK组的茜素红染色程度也为1。比fRunx2-Tat组高2倍。fRunx2-cLK组的ALP和骨钙素基因表达水平高于其他组。fRunx2在cLK肽的辅助下有效转移到细胞质中，并增强了MSC的成骨分化。
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