Solovyeva Valeriya V, Kitaeva Kristina V, Chulpanova Daria S, Arkhipova Svetlana S, Filin Ivan Yu, Rizvanov Albert A
Kazan Federal University, Kazan, Russia.
Bionanoscience. 2022;12(2):293-301. doi: 10.1007/s12668-021-00931-5. Epub 2022 Mar 4.
At present, there is an increasing interest in the potential role of extracellular vesicles (EVs), acting as multi-signal messengers of the tumor stroma, in the development and progression of tumor. Tumor cell-derived EVs are considered a potential vector for the targeted delivery of antitumor agents due to the ability to fuse with parental cells through endocytosis and release their contents into the cytoplasm of the recipient cell. Tumor cell-derived EVs could be also used for priming immune cells and therapeutic vaccine development. It is also known that mesenchymal stem cells (MSCs) have a tropism toward tumor niches. It is believed that MSC migration to the tumor is due to its inflammatory signaling. Presumably, with the accumulation of MSCs at tumor sites, these cells differentiate into pericytes or tumor-associated fibroblasts, thereby forming a supporting tumor growth microenvironment. However, besides the ability to promote tumor progression, MSCs can also suppress its growth by inhibiting proliferation and cell cycle progression, and angiogenesis. Thus, the further studies of the MSC role in TME and MSC interaction with other cells of the tumor stroma, including through EVs, are of particular interest. To increase the yield of vesicles the isolation method based on pharmacological disorganization of the actin cytoskeleton induced by treating with cytochalasin B was used in this study. In this investigation the interaction of SH-SY5Y neuroblastoma cell-derived membrane vesicles, obtained using cytochalasin B (CIMVs), with human bone marrow-derived MSCs was analyzed using imaging flow cytometry. Using transmission electron microscopy, it was shown that CIMVs have a size similar to that of natural microvesicles, which is 100-1000 nm. Using imaging flow cytometry, it was shown that after 24 h of co-cultivation 6% of the MSCs contained a large number of CIMVs, and 42% of the MSCs contained a small amount of CIMVs. Cultivation of MSCs with SH-SY5Y cell-derived CIMVs also induced dose-dependent decrease in the expression of CD markers typical for MSCs. Thus, the internalization of SH-SY5Y cell-derived CIMVs within MSCs and the ability of the CIMVs to modulate immunophenotype of the recipient cells were shown. However, further studies are required to determine the effect of CIMVs on pro- or antioncogenic phenotype and function of MSCs.
目前，细胞外囊泡（EVs）作为肿瘤基质的多信号信使在肿瘤发生发展中的潜在作用越来越受到关注。肿瘤细胞衍生的EVs被认为是抗肿瘤药物靶向递送的潜在载体，因为它们能够通过内吞作用与亲代细胞融合并将其内容物释放到受体细胞的细胞质中。肿瘤细胞衍生的EVs还可用于启动免疫细胞和开发治疗性疫苗。此外，已知间充质干细胞（MSCs）对肿瘤微环境具有趋向性。据信，MSCs向肿瘤的迁移归因于其炎症信号。据推测，随着MSCs在肿瘤部位的积累，这些细胞分化为周细胞或肿瘤相关成纤维细胞，从而形成支持肿瘤生长的微环境。然而，除了促进肿瘤进展的能力外，MSCs还可通过抑制增殖、细胞周期进程和血管生成来抑制肿瘤生长。因此，进一步研究MSCs在肿瘤微环境中的作用以及MSCs与肿瘤基质中其他细胞的相互作用，包括通过EVs的相互作用，具有特别重要的意义。为了提高囊泡产量，本研究采用了基于细胞松弛素B处理诱导肌动蛋白细胞骨架药理学解体的分离方法。在本研究中，使用成像流式细胞术分析了用细胞松弛素B获得的SH-SY5Y神经母细胞瘤细胞衍生的膜囊泡（CIMVs）与人骨髓来源的MSCs之间的相互作用。使用透射电子显微镜显示，CIMVs的大小与天然微囊泡相似，为100-1000nm。使用成像流式细胞术显示，共培养24小时后，6%的MSCs含有大量CIMVs，42%的MSCs含有少量CIMVs。用SH-SY5Y细胞衍生的CIMVs培养MSCs也导致MSCs典型的CD标志物表达呈剂量依赖性下降。因此，显示了SH-SY5Y细胞衍生的CIMVs在MSCs内的内化以及CIMVs调节受体细胞免疫表型的能力。然而，需要进一步研究以确定CIMVs对MSCs促癌或抑癌表型及功能的影响。
