PO.TB10.15 · 肿瘤生物学
探究cofilin-1过表达细胞分泌的细胞外囊泡对人肺癌细胞迁移和侵袭能力的影响
Exploring the effects of extracellular vesicles secreted by cofilin-1 overexpressing cells on the migration and invasion ability of human lung cancer cells
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摘要 Abstract
中文摘要
目的：I型肌动蛋白切割蛋白cofilin-1（CFL-1）是非肌细胞中广泛表达的微丝细胞骨架调节因子。它在肌动蛋白细胞骨架构建中发挥关键作用，并影响细胞运动能力。CFL-1的过表达已在多种癌症中被观察到，并与肿瘤的迁移、侵袭和转移相关。此外，研究表明细胞骨架在胞吐过程中发挥关键作用，提示CFL-1可能是该过程的潜在调节因子。癌细胞分泌大量细胞外囊泡（EVs）以介导细胞间通讯，而外泌体是其亚型之一。肿瘤来源的外泌体可促进肿瘤侵袭和转移。近期研究显示，CFL-1在晚期癌症的外泌体中高表达，但其在肿瘤相关外泌体中的潜在机制仍不明确。因此，本研究旨在探究CFL-1对外泌体分泌的影响，以及CFL-1是否通过外泌体介导的机制影响肺癌细胞。
方法：本研究使用H1299/tet-on-cofilin-1（HCOXP）细胞，这是一种四环素诱导型细胞系，可诱导cofilin-1过表达。首先，从过表达cofilin-1的HCOXP细胞收集条件培养基，并通过超速离心分离外泌体。其次，通过纳米颗粒追踪分析、透射电子显微镜和Western blot验证所分离外泌体的纯度。为评估对细胞迁移和侵袭的影响，将HCOXP细胞来源的外泌体加入H1299、CL1-0和BEAS2-B细胞。此外，比较了过表达和未过表达cofilin-1的HCOXP细胞外泌体中基质金属蛋白酶（MMPs）的表达水平。
结果：结果显示，CFL-1过表达刺激细胞分泌更多外泌体。同时也发现这些外泌体可能含有更多内容物。当将这些外泌体应用于肺癌细胞和正常细胞时，来源于CFL-1过表达HCOXP细胞的外泌体显著加速了H1299细胞的迁移和侵袭。然而，在经这些外泌体处理后，CL1-0或BEAS-2B细胞并未观察到迁移和侵袭能力的增加。此外，在CFL-1过表达HCOXP细胞的外泌体中检测到基质金属蛋白酶（MMPs）水平升高。
结论：研究发现HCOXP细胞中CFL-1的过表达可促进外泌体分泌。这些外泌体似乎表现出归巢特异性，可能影响其对靶细胞的作用。总体而言，本研究揭示了CFL-1在调控外泌体释放中的关键作用及其在肿瘤微环境中的潜在影响。
查看英文原文 English abstract
Object: Type I actin-severing protein cofilin-1 (CFL-1) is a widely expressed regulator of the microfilament cytoskeleton in non-muscle cells. It plays a crucial role in actin cytoskeleton construction and influences cell motility. Overexpression of CFL-1 has been observed in various cancers and is related to tumor migration, invasion, and metastasis. In addition, studies have shown that the cytoskeleton plays a key role in exocytosis, implicating that CFL-1 may be a potential regulator of this process. Cancer cells secrete abundant extracellular vesicles (EVs) to mediate intercellular communication, and exosomes are one of the subtypes. Tumor-derived exosomes can promote tumor invasion and metastasis. Recent studies showed that CFL-1 is highly expressed in exosomes from advanced cancer, but its potential mechanisms in tumor-related exosomes remain unclear. Therefore, this study aims to explore the effects of CFL-1 on exosome secretion and whether CFL-1 influences lung cancer cells via exosome-mediated mechanisms.
Methods: This study used H1299/tet-on-cofilin-1 (HCOXP) cells, a tetracycline-inducible cell line, which can induce cofilin-1 overexpression. First, conditioned medium was collected from HCOXP cells overexpressing cofilin-1, and exosomes were isolated by ultracentrifugation. Second, the purity of the isolated exosomes was verified by nanoparticle tracking analysis, transmission electron microscopy, and western blotting. To evaluate the impact on cell migration and invasion, exosomes derived from HCOXP cells were added to H1299, CL1-0, and BEAS2-B cells. Furthermore, the expression levels of matrix metalloproteinases (MMPs) in exosomes from HCOXP cells with and without cofilin-1 overexpression were compared.
Results: The results showed that overexpression of CFL-1 stimulated cells to secrete more exosomes. It is also found that these exosomes may contain more contents. When these exosomes were applied to lung cancer and normal cells, exosomes derived from CFL-1 overexpressing HCOXP cells significantly accelerated the migration and invasion of H1299 cells. However, no increase in migration and invasion ability was observed in CL1-0 or BEAS-2B cells following treatment with these exosomes. In addition, elevated levels of matrix metalloproteinases (MMPs) were detected in exosomes from CFL-1-overexpressing HCOXP cells.
Conclusion: Overexpression of CFL-1 in HCOXP cells was found to promote exosome secretion. These exosomes appear to exhibit homing specificity, potentially influencing their effects on target cells. Overall, this study found the critical role of CFL-1 in regulating the release and the potential impact of exosomes in the tumor microenvironment.
利益披露 Disclosure
H. Bo-Han, None..
Y. Lee, None.