含 MiR-29a 的细胞外囊泡通过阻断自噬作用有效诱导 HCC 细胞凋亡。

IF 4.4 2区 医学 Q1 PHARMACOLOGY & PHARMACY
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引用次数: 0

摘要

背景:尽管肝细胞癌(HCC)的治疗方法取得了重大进展,但每年的死亡率仍然很高,而且发病率还在不断上升。HCC 临床治疗面临的主要挑战与耐药性的产生和肿瘤细胞凋亡的逃避有关,这导致 HCC 患者的治疗效果不尽人意。以往的研究表明,自噬在 HCC 的耐药性发展中起着至关重要的作用。尽管已知 miR-29a 能抵消自噬作用,但越来越多的证据表明,在 HCC 患者中,miR-29a 的下调与不良预后相关。此外,还有证据表明,miR-29a 在其他癌症中也是自噬的负调控因子。在本研究中,我们旨在利用细胞外囊泡(EVs)作为一种天然递送系统,研究 miR-29a 对 HCC 细胞自噬和凋亡的影响,因为它们在体外和体内都具有递送 miRNA 的潜力:方法:通过20,000或110,000×g离心(分别为EV20K或EV110K),最近分离出了人Wharton's Jelly间充质基质细胞衍生的细胞外囊泡,并通过Western印迹(WB)、扫描电子显微镜(SEM)和动态光散射(DLS)对其进行了表征。随后将miR-29a载入这些EVs,并通过RT-qPCR评估其载入效率。然后对 Huh-7 和 HepG2 细胞系进行了全面的体外和体内评估:结果:在 Huh-7 和 HepG2 细胞系中,EV20K-miR-29a 处理可显著诱导细胞凋亡,并减少细胞增殖和集落形成。此外,LC3-II/LC3-I 比值升高,而关键自噬调节因子 TFEB 和 ATG9A 的表达在该处理中下调。这些研究结果表明,EV20K-miR-29a 通过同时靶向每种途径中的关键介质,有效阻断了自噬,从而导致了 HCC 细胞株的凋亡。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

MiR-29a-laden extracellular vesicles efficiently induced apoptosis through autophagy blockage in HCC cells

MiR-29a-laden extracellular vesicles efficiently induced apoptosis through autophagy blockage in HCC cells

Background

In spite of significant advancements in theraputic modalities for hepatocellular carcinoma (HCC), there is still a high annual mortality rate with a rising incidence. Major challenges in the HCC clinical managment are related to the development of therapy resistance, and evasion of tumor cells apoptosis which leading unsatisfactory outcomes in HCC patients. Previous investigations have shown that autophagy plays crucial role in contributing to drug resistance development in HCC. Although, miR-29a is known to counteract authophagy, increasing evidence revealed a down-regulation of miR-29a in HCC patients which correlates with poor prognosis. Beside, evidences showed that miR-29a serves as a negative regulator of autophagy in other cancers. In the current study, we aim to investigate the impact of miR-29a on the autophagy and apoptosis in HCC cells using extracellular vesicles (EVs) as a natural delivery system given their potential in the miRNA delivery both in vitro and in vivo.

Method

Human Wharton’s Jelly mesenchymal stromal cell-derived extracellular vesicles were lately isolated through 20,000 or 110,000 × g centrifugation (EV20K or EV110K, respectively), characterized by western blot (WB), scanning electron microscopy (SEM), and dynamic light scattering (DLS). miR-29a was subsequently loaded into these EVs and its loading efficiency was evaluated via RT-qPCR. Comprehensive in vitro and in vivo assessments were then performed on Huh-7 and HepG2 cell lines.

Results

EV20K-miR-29a treatment significantly induces cell apoptosis and reduces both cell proliferation and colony formation in Huh-7 and HepG2 cell lines. In addition, LC3-II/LC3-I ratio was increased while the expression of key autophagy regulators TFEB and ATG9A were downregulated by this treatment. These findings suggest an effective blockade of autophagy by EV20K-miR-29a leading to apoptosis in the HCC cell lines through concomitant targeting of critical mediators within each pathway.

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来源期刊
CiteScore
8.80
自引率
4.10%
发文量
211
审稿时长
36 days
期刊介绍: The European Journal of Pharmaceutics and Biopharmaceutics provides a medium for the publication of novel, innovative and hypothesis-driven research from the areas of Pharmaceutics and Biopharmaceutics. Topics covered include for example: Design and development of drug delivery systems for pharmaceuticals and biopharmaceuticals (small molecules, proteins, nucleic acids) Aspects of manufacturing process design Biomedical aspects of drug product design Strategies and formulations for controlled drug transport across biological barriers Physicochemical aspects of drug product development Novel excipients for drug product design Drug delivery and controlled release systems for systemic and local applications Nanomaterials for therapeutic and diagnostic purposes Advanced therapy medicinal products Medical devices supporting a distinct pharmacological effect.
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