Kim Boyong
EVERBIO, 131, Jukhyeon-gil, Gwanghyewon-myeon, Jincheon-gun 27809, Republic of Korea.
Int J Mol Sci. 2024 Dec 11;25(24):13299. doi: 10.3390/ijms252413299.
The increasing incidence and mortality rates of liver cancer have heightened the demand for the development of effective anticancer drugs with minimal side effects. In this study, the roles of exosomes derived from liver cancer stem cells (LCSCs) with PRELI (Protein of Relevant Evolutionary and Lymphoid Interest) modulation and their miRNAs were investigated to explore their therapeutic properties for liver cancer. Various techniques, such as miRNA profiling, microRNA transfection, overexpression, flow cytometry, Western blotting, and immunocytochemistry, were used to evaluate the effects of exosomes under PRELI up- and downregulation. Downregulated PRELI cellular exosomes (DPEs) reduced the levels of five markers-CD133, CD90, CD24, CD13, and EpCAM-in LCSCs, with the exception of OV-6. Conversely, upregulated PRELI cellular exosomes (UPEs) significantly increased the expression of CD90, CD24, and CD133 in NHs, with the maximum increase in CD24. PRELI upregulation altered expression levels of miRNAs, including hsa-miR-378a-3p (involved in stem-like properties), hsa-miR-25-3p (contributing to cell proliferation), and hsa-miR-423-3p (driving invasiveness). Exosomes with downregulated PRELI inhibited the AKT/mTORC1 signaling pathway, whereas LCSCs transfected with the candidate miRNAs activated it. Additionally, under PRELI upregulation, exosomes showed increased surface marker expression, promoting cancer progression. The modulation of PRELI in LCSCs affected miRNA expression significantly, revealing candidate miRNA targets for liver cancer treatment. Exosomes with PRELI downregulation show potential as a novel therapeutic strategy. Consequently, this study proposes the potential of PRELI-induced exosomes and the three miRNAs as a liver anticancer therapeutic candidate.
肝癌发病率和死亡率的不断上升，增加了对开发副作用最小的有效抗癌药物的需求。在本研究中，我们研究了具有PRELI（相关进化和淋巴样蛋白）调节作用的肝癌干细胞（LCSCs）来源的外泌体及其miRNA的作用，以探索它们对肝癌的治疗特性。我们使用了多种技术，如miRNA谱分析、微小RNA转染、过表达、流式细胞术、蛋白质印迹法和免疫细胞化学，来评估PRELI上调和下调情况下外泌体的作用。下调PRELI的细胞外泌体（DPEs）降低了LCSCs中五个标志物——CD133、CD90、CD24、CD13和EpCAM的水平，但OV-6除外。相反，上调PRELI的细胞外泌体（UPEs）显著增加了正常肝细胞（NHs）中CD90、CD24和CD133的表达，其中CD24的增加最为明显。PRELI上调改变了miRNA的表达水平，包括hsa-miR-378a-3p（参与干细胞样特性）、hsa-miR-25-3p（促进细胞增殖）和hsa-miR-423-3p（驱动侵袭性）。PRELI下调的外泌体抑制了AKT/mTORC1信号通路，而用候选miRNA转染的LCSCs激活了该信号通路。此外，在PRELI上调的情况下，外泌体显示出表面标志物表达增加，促进了癌症进展。LCSCs中PRELI的调节显著影响了miRNA的表达，揭示了肝癌治疗的候选miRNA靶点。PRELI下调的外泌体显示出作为一种新型治疗策略的潜力。因此，本研究提出了PRELI诱导的外泌体和这三种miRNA作为肝脏抗癌治疗候选物的潜力。
