背景:尽管对间充质干细胞(MSC)感兴趣,他们治疗异常疤痕的潜力,尤其是瘢痕疙瘩,还有待描述。本研究旨在研究源自人骨髓MSCs(hBMSC-Exos)的外泌体在减轻瘢痕疙瘩形成中的治疗潜力。
方法:从hBMSC中分离外泌体,和瘢痕疙瘩成纤维细胞(KF)用hBMSC-Exos处理。细胞计数试剂盒-8,伤口愈合,Transwell入侵,免疫荧光,并进行蛋白质印迹分析以研究KFs的恶性表型。用瘢痕疙瘩诱导小鼠并用hBMSC-Exos处理。通过苏木精和伊红染色评估hBMSC-Exos对体内瘢痕疙瘩形成的影响,Masson染色,免疫组织化学,和西方印迹。针对瘢痕疙瘩形成期间差异表达的长非编码RNA,筛选GSE182192数据集。接下来,在hBMSC中敲低母体表达的基因3(MEG3)以获得hBMSC-Exossh-MEG3。通过生物信息学筛选研究了MEG3的分子机制,并验证了MEG3与TP53或MCM5的关系。
结果:hBMSC-Exos抑制恶性增殖,迁移,和KFs的侵袭,同时促进其凋亡,此外,hBMSC-Exos降低了细胞中纤维化和胶原相关蛋白的表达以及由KFs引起的瘢痕疙瘩的形成。hBMSC-Exos中MEG3富集的减少削弱了hBMSC-Exos对KF活性的抑制作用。hBMSC-Exos通过TP53递送MEG3以促进KF中的MCM5转录。MCM5在KF中的过表达逆转了hBMSC-Exossh-MEG3的作用,导致KF活性降低。
结论:hBMSC-Exos递送MEG3以促进TP53的蛋白质稳定性,从而激活MCM5并促进KF活性。
BACKGROUND: Despite the interest in mesenchymal stem cells (MSC), their potential to treat abnormal scarring, especially keloids, is yet to be described. The present study aimed to investigate the therapeutic potential of exosomes derived from human bone marrow MSCs (hBMSC-Exos) in alleviating keloid formation.
METHODS: Exosomes were isolated from hBMSC, and keloid fibroblasts (KFs) were treated with hBMSC-Exos. Cell counting kit-8, wound healing, transwell invasion, immunofluorescence, and western blot assays were conducted to study the malignant phenotype of KFs. Mice were induced with keloids and treated with hBMSC-Exos. The effect of hBMSC-Exos on keloid formation in vivo was evaluated by hematoxylin and eosin staining, Masson staining, immunohistochemistry, and western blotting. The GSE182192 dataset was screened for differentially expressed long non-coding RNA during keloid formation. Next, maternally expressed gene 3 (
MEG3) was knocked down in hBMSC to obtain hBMSC-Exossh-
MEG3. The molecular mechanism of
MEG3 was investigated by bioinformatic screening, and the relationship between
MEG3 and TP53 or MCM5 was verified.
RESULTS: hBMSC-Exos inhibited the malignant proliferation, migration, and invasion of KFs at same time as promoting their apoptosis, Moreover, hBMSC-Exos reduced the expression of fibrosis- and collagen-related proteins in the cells and the formation of keloids caused by KFs. The reduction in MEG3 enrichment in hBMSC-Exos weakened the inhibitory effect of hBMSC-Exos on KF activity. hBMSC-Exos delivered
MEG3 to promote MCM5 transcription by TP53 in KFs. Overexpression of MCM5 in KFs reversed the effects of hBMSC-Exossh-
MEG3, leading to reduced KF activity.
CONCLUSIONS: hBMSC-Exos delivered MEG3 to promote the protein stability of TP53, thereby activating MCM5 and promoting KF activity.