Institute for Human Tissue Restoration, Department of Plastic & Reconstructive Surgery, Yonsei University College of Medicine, Seoul, Korea.
Department of Bioengineering, College of Engineering, Hanyang University, 222 Wangsimni-ro, Seongdong-gu, Seoul, 04763, Korea.
Sci Rep. 2017 Oct 11;7(1):12957. doi: 10.1038/s41598-017-13485-y.
Mortalin (Mot) is a mitochondrial chaperone of the heat shock protein 70 family and it's pro-proliferative and anti-apoptosis functions could be associated with keloid pathogenesis, and blocking of mortalin and its interaction with p53 might be a potential novel target for the treatment of keloid. Therefore, we generated mortalin-specific small hairpin (sh) RNAs (dE1-RGD/GFP/shMot) and introduced into keloid spheroids for examination of its apoptotic and anti-fibrotic effect. On keloid tissues, mortalin expression was higher than adjacent normal tissues and it's protein expressions were activated keloid fibroblasts (KFs). After primary keloid spheroid were transduced with dE1-RGD/GFP/shMot for knockdown of mortalin, expression of type I, III collagen, fibronectin, and elastin was significantly reduced and transforming growth factor-β1, epidermal growth factor receptor (EGFR), Extracellular Signal-Regulated Kinases 1 and 2 (Erk 1/2), and Smad 2/3 complex protein expression were decreased. In addition, increased TUNEL activities and cytochrome C were observed. Further, for examine of mortalin and p53 interaction, we performed immunofluorescence analysis. Knockdown of mortalin relocated p53 to the cell nucleus in primary keloid spheroids by dE1-RGD/GFP/shMot transduction. These results support the utility of knockdown of mortalin to induce apoptosis and reduce ECMs expression in keloid spheroid, which may be highly beneficial in treating keloids.
线粒体蛋白（Mot）是热休克蛋白 70 家族的一种线粒体伴侣，其促有丝分裂和抗细胞凋亡功能可能与瘢痕疙瘩的发病机制有关，阻断 Mot 及其与 p53 的相互作用可能是瘢痕疙瘩治疗的一个新靶点。因此，我们生成了 Mot 特异性短发夹（sh）RNA（dE1-RGD/GFP/shMot），并将其导入瘢痕疙瘩球体中，以研究其促凋亡和抗纤维化作用。在瘢痕疙瘩组织中，Mot 的表达高于相邻的正常组织，其蛋白表达在激活的瘢痕疙瘩成纤维细胞（KFs）中被激活。在原代瘢痕疙瘩球体中转导 dE1-RGD/GFP/shMot 以敲低 Mot 后，I 型、III 型胶原、纤维连接蛋白和弹性蛋白的表达显著降低，转化生长因子-β1、表皮生长因子受体（EGFR）、细胞外信号调节激酶 1 和 2（Erk 1/2）和 Smad 2/3 复合物蛋白的表达也降低。此外，还观察到 TUNEL 活性和细胞色素 C 的增加。进一步，为了检测 Mot 和 p53 的相互作用，我们进行了免疫荧光分析。通过 dE1-RGD/GFP/shMot 转导，敲低 Mot 将 p53 重定位到原代瘢痕疙瘩球体的细胞核中。这些结果支持了敲低 Mot 诱导凋亡和减少 ECMs 在瘢痕疙瘩球体中的表达的作用，这可能对治疗瘢痕疙瘩非常有益。
