人骨髓间充质干细胞来源的外泌体运输的 MEG3 通过稳定 TP53 调节瘢痕成纤维细胞中 MCM5 的转录。

MEG3 shuttled by exosomes released from human bone marrow mesenchymal stem cells promotes TP53 stability to regulate MCM5 transcription in keloid fibroblasts.

机构信息

Department of Burn Surgery, Huizhou Central People's Hospital, Huizhou, China.

Hand and Foot Microsurgery & Wound Repair Department, Huizhou First Hospital, Huizhou, China.

出版信息

J Gene Med. 2024 May;26(5):e3688. doi: 10.1002/jgm.3688.

DOI:10.1002/jgm.3688

PMID:38686583

Abstract

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-Exos. 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-Exos, leading to reduced KF activity.

CONCLUSIONS

hBMSC-Exos delivered MEG3 to promote the protein stability of TP53, thereby activating MCM5 and promoting KF activity.

摘要

背景

尽管间充质干细胞（MSC）备受关注，但它们在治疗异常瘢痕，特别是瘢痕疙瘩方面的潜力尚未得到描述。本研究旨在探讨人骨髓间充质干细胞（hBMSC）来源的外泌体（hBMSC-Exos）在缓解瘢痕疙瘩形成中的治疗潜力。

方法

从 hBMSC 中分离外泌体，并将其用于处理瘢痕疙瘩成纤维细胞（KFs）。通过细胞计数试剂盒-8、划痕愈合、Transwell 侵袭、免疫荧光和 Western blot 实验来研究 KFs 的恶性表型。通过诱导小鼠形成瘢痕疙瘩并用 hBMSC-Exos 进行治疗，评估 hBMSC-Exos 对体内瘢痕疙瘩形成的影响。通过苏木精和伊红染色、Masson 染色、免疫组化和 Western blot 对 hBMSC-Exos 在体内形成瘢痕疙瘩的效果进行评估。通过筛选 GSE182192 数据集来研究瘢痕疙瘩形成过程中差异表达的长链非编码 RNA。接着，在 hBMSC 中敲低母系表达基因 3（MEG3）以获得 hBMSC-Exos。通过生物信息学筛选来研究 MEG3 的分子机制，并验证 MEG3 与 TP53 或 MCM5 的关系。

结果

hBMSC-Exos 同时抑制 KFs 的恶性增殖、迁移和侵袭，并促进其凋亡。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 KFs 引起的瘢痕疙瘩形成。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 将 MEG3 递送至 KFs 中，从而促进 MCM5 的转录。在 KFs 中转染 MCM5 过表达可逆转 hBMSC-Exos 的作用，导致 KF 活性降低。