截短型eIF3e基因治疗促皮肤创面愈合及分子机制探究

eIF3e plays an important role in the initiation and regulation of eukaryotic translation. Downregulation of its expression can enhance the stability of HIF-2α and induce EMT to promote wound healing. In the study of murine breast cancer, it was found that after MMTV insertion into eIF3e gene, a variety of negative truncated mutants can be produced. However, whether these mutants can promote wound healing has not been reported yet. Our preliminary experiments showed that truncated eIF3e-C and eIF3e-D promote wound healing through promoting epithelial and granulation tissue formation, but no obvious enhancement of vascularization and extracellular matrix deposition were observed. The mechanism was unknown. This project intends to explore its effect on the biological behavior of cell proliferation, migration, apoptosis, EMT, and the expression of eIF4E phosphorylation and the related proteins transfected by different truncated eIF3e mutants, eIF3d and c-Jun. Mice full-thickness skin defect model was constructed to evaluate eIF3e, its truncated mutants, eIF3d and c-Jun in wound healing. In order to explore the molecular mechanism of truncated eIF3e in wound healing, and promote the establishment of efficient new strategies to provide experimental basis for wound healing.

eIF3e在真核翻译起始和调控蛋白水平中发挥重要作用，下调其表达能通过提高HIF-2α稳定性及诱发EMT促进创面愈合。在对小鼠乳腺癌研究中发现MMTV插入eIF3e基因后可产生多种显负性的截短型突变体，然而这些突变体是否能促进创面愈合尚无报道。我们的前期研究表明，截短型eIF3e-C和eIF3e-D能够通过促再上皮化和肉芽组织形成促进创面愈合，但并未观察到明显的血管化和细胞外基质沉积增强，其机制不明。本项目拟通过构建不同截短型eIF3e突变体、eIF3d和c-Jun真核表达载体系统探究其对细胞增殖、迁移、凋亡和EMT等生物学行为的影响，同时检测其对相关蛋白表达水平和eIF4E磷酸化的影响，在此基础上使用小鼠皮肤全层缺损模型探究eIF3e及其截短型突变体在创面愈合中的作用，并初步揭示其促创面愈合的分子机制，从而为建立高效促创面愈合的新策略提供实验依据。

皮肤作为人体最大的器官，不仅是有机体抵御外界刺激和病原微生物侵袭的第一道防线，而且具有感知冷热和压力等功能。每年，急性和慢性创面影响着数以千万人，据估计，每年有超过250亿美元用在治疗慢性创面上。随着人口老龄化、全球性糖尿病及肥胖发病率的增加，该项治疗费用急剧上涨。因此研发促进创面愈合的新靶点和新策略仍是该领域亟需解决的关键问题。eIF3e在真核翻译起始和调控蛋白水平中发挥重要作用，下调其表达能通过提高HIF-2α稳定性及诱发EMT促进创面愈合。在对小鼠乳腺癌研究中发现MMTV插入eIF3e基因后可产生多种显负性的截短型突变体，然而这些突变体是否能促进创面愈合尚无报道。本项目拟通过构建不同截短型eIF3e突变体、eIF3d和c-Jun表达载体系统探究其对细胞增殖、迁移、凋亡和EMT等生物学行为及相关蛋白表达水平的影响，在此基础上使用小鼠皮肤全层缺损模型探究eIF3e及其截短型突变体在创面愈合中的作用，并初步揭示其促创面愈合的分子机制。研究结果表明，eIF3e突变体促进细胞增殖，通过促进再上皮化和肉芽组织形成促进创面愈合。通过本研究，我们构建了一种可实现在体局部转染促创面愈合基因的治疗方法，为建立高效促创面愈合的新策略提供了实验依据。