人脐带间充质干细胞分泌的外泌体通过转移 miR-100-5p 促进胰腺导管腺癌生长。
Exosomes secreted from human umbilical cord mesenchymal stem cells promote pancreatic ductal adenocarcinoma growth by transferring miR-100-5p.
机构信息
Department of General Surgery, Xuanwu Hospital, Capital Medical University, Beijing, 100053, China; Clinical Center for Acute Pancreatitis, Capital Medical University, Beijing, 100053, China.
Department of General Surgery, Xuanwu Hospital, Capital Medical University, Beijing, 100053, China.
出版信息
Tissue Cell. 2021 Dec;73:101623. doi: 10.1016/j.tice.2021.101623. Epub 2021 Aug 14.
PURPOSE
Although human umbilical cord mesenchymal stem cells (hucMSCs) can contribute to the growth of tumors, including pancreatic ductal adenocarcinoma (PDAC), however, little is known about the exact mechanisms by which the exosomes secreted from hucMSCs (hucMSCs-exo) have an oncogenic effect on the physiopathology of PDAC. The effects of hucMSCs on tumor development are attributed to hucMSCs-exo, which deliver unique proteins and miRNAs to cancer cells.
METHODS
HucMSCs and exosomes were isolated and confirmed via transmission electron microscopy, nanoparticle tracking analysis and western blot. The nude mice were inoculated subcutaneously on both flanks with human pancreatic cancer Panc-1 cells (1 × 10), and hucMSCs-exo were directly administered via intratumoral injection once a day for three days each week. Cell proliferation assays were performed using a Cell Counting Kit-8 assay and the cell invasion assay was performed using Transwell assay. The miRNA data were predicted and analyzed by miRanda software. The analysis of the target genes of the miRNAs was proformed with the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases.
RESULTS
Firstly, we observed that hucMSCs-exo promoted Panc-1 and BxPC3 cell growth by increasing proliferation and migration in vitro. Secondly, in a xenograft tumor model, hucMSCs-exo increased the growth of Panc-1 cells. Thirdly, high-throughput sequencing of hucMSCs-exo showed that hsa-miR-148a-3p, hsa-miR-100-5p, hsa-miR-143-3p, hsa-miR-21-5p and hsa-miR-92a-3p were highly expressed. For the five identified miRNAs, 1308 target genes were predicted by miRanda software. From the GO and KEGG analyses of the target genes of the identified miRNAs, it was found that the main GO function was the regulation of cellular glucuronidation, and the main KEGG metabolic pathway involved the metabolism of ascorbic acid and aldehyde acid. These processes are related to the occurrence and development of pancreatic cancer. Finally, we observed that miR-100-5p promoted Panc-1 and BxPC3 cell growth in vitro and in vivo.
CONCLUSION
Here, by utilizing exosomes secreted from hucMSCs, we systematically investigated the effects of hucMSCs-exo on PDAC growth in vitro and in vivo for the first time. Building on these results, we provided new insights into the role of hucMSCs-exo in the PDAC growth and revealed the attractive communication between hucMSCs and PDAC cells that occurs through MSCs-exosomes-miRNAs.
目的
人脐带间充质干细胞（hucMSCs）可促进肿瘤生长，包括胰腺导管腺癌（PDAC），然而，hucMSCs 分泌的外泌体（hucMSCs-exo）对 PDAC 病理生理学的确切机制知之甚少。hucMSCs 对肿瘤发展的影响归因于 hucMSCs-exo，它将独特的蛋白质和 miRNAs 递送到癌细胞中。
方法
通过透射电子显微镜、纳米颗粒跟踪分析和 Western blot 分离和确认 hucMSCs 和外泌体。将人胰腺癌细胞 Panc-1（1×10）接种到裸鼠双侧皮下，每周三次每天一次通过肿瘤内注射直接给予 hucMSCs-exo。使用细胞计数试剂盒-8 测定进行细胞增殖测定，使用 Transwell 测定进行细胞侵袭测定。通过 miRanda 软件预测和分析 miRNA 数据。使用基因本体论（GO）和京都基因与基因组百科全书（KEGG）数据库分析 miRNA 的靶基因。
结果
首先，我们观察到 hucMSCs-exo 通过增加体外增殖和迁移促进 Panc-1 和 BxPC3 细胞生长。其次，在异种移植肿瘤模型中，hucMSCs-exo 增加了 Panc-1 细胞的生长。第三，hucMSCs-exo 的高通量测序显示 hsa-miR-148a-3p、hsa-miR-100-5p、hsa-miR-143-3p、hsa-miR-21-5p 和 hsa-miR-92a-3p 高度表达。通过 miRanda 软件预测，这 5 个鉴定的 miRNAs 有 1308 个靶基因。从鉴定 miRNA 的靶基因的 GO 和 KEGG 分析中发现，主要的 GO 功能是细胞葡萄糖醛酸化的调节，主要的 KEGG 代谢途径涉及抗坏血酸和醛酸的代谢。这些过程与胰腺癌的发生和发展有关。最后，我们观察到 miR-100-5p 促进了体外和体内的 Panc-1 和 BxPC3 细胞生长。
结论
在这里，我们首次利用 hucMSCs 分泌的外泌体，系统研究了 hucMSCs-exo 对 PDAC 体外和体内生长的影响。在此基础上，我们为 hucMSCs-exo 在 PDAC 生长中的作用提供了新的见解，并揭示了 hucMSCs 与 PDAC 细胞之间通过 MSC-exosomes-miRNAs 进行的有吸引力的交流。
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