tam来源的外泌体miR-589-3p通过BCL2L13加速卵巢癌进展。

IF 3.8 3区 医学 Q1 REPRODUCTIVE BIOLOGY
Jianqing Wang, Yan Zhu, Yang He, Weiwei Shao
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引用次数: 0

摘要

背景:肿瘤相关巨噬细胞(tumor associated macrophages, TAM)是肿瘤微环境(tumor microenvironment, TME)中细胞间通讯的关键元件,而外泌体是肿瘤细胞与TME之间的关键介质。根据先前的报道,mirna在卵巢癌(OC)的发展中发挥着关键作用。这项工作的目的是探讨tam来源的外泌体miR-589-3p在OC发育中的功能,并阐明其潜在的分子机制。方法:首先用IL-4和IL-13处理外周血单核细胞(PBMC),使其极化为m2型巨噬细胞;从m2型巨噬细胞中分离外泌体,利用透射电镜(TEM)和纳米颗粒跟踪分析(NTA)对外泌体的物理性质进行评价。接下来,应用定量逆转录酶聚合酶链反应(qRT-PCR)检测相关基因的表达。随后,利用Targetscan和miRDB预测miR-589-3p靶基因,然后通过双荧光素酶法和RNA结合蛋白免疫沉淀(RIP)法验证miR-589-3p与BCL2L13的相互作用。最后通过细胞计数试剂盒-8 (CCK-8)和流式细胞术实验探讨OC细胞增殖和凋亡能力的变化。结果:在本研究中,我们证明tam来源的外泌体促进OC细胞增殖并抑制OC细胞凋亡。然后,qRT-PCR结果显示,tam来源的外泌体与OC细胞共培养后,miR-589-3p明显升高。此外,我们发现miR-589-3p与bcl -2样蛋白13 (BCL2L13)结合,通过荧光素酶实验和RIP实验证实了这一点。此外,功能分析显示,用miR-589-3p抑制剂处理tam衍生的外泌体减弱了外泌体对OC细胞进展的促进作用。结论:tam来源的外泌体miR-589-3p通过BCL2L13促进OC进展,为OC治疗提供了一种新的方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
TAM-derived exosomal miR-589-3p accelerates ovarian cancer progression through BCL2L13.

Background: Tumor-associated macrophages (TAM) are critical elements of intercellular communication in tumor microenvironment (TME), and exosomes are key mediators between tumor cells and the TME. According to previous reports, miRNAs exert a pivotal role in ovarian cancer (OC) development. The purpose of this work was to explore the function of TAM-derived exosomal miR-589-3p in OC development and elucidate the underlying molecular mechanisms.

Methods: First, peripheral blood mononuclear cells (PBMC) were treated with IL-4 and IL-13 to polarize them into M2-type macrophages. Exosomes were separated from M2-type macrophages, and the physical properties of exosomes were evaluated using transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA). Next, quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) was applied to examine the expression of relevant genes. Subsequently, Targetscan and miRDB were utilized to predict miR-589-3p target genes, and then the interaction between miR-589-3p and BCL2L13 was verified by dual luciferase assay and RNA Binding Protein Immunoprecipitation (RIP) assay. Finally, Cell Counting Kit-8 (CCK-8) and flow cytometry experiments were employed to explore the changes in the proliferative and apoptotic abilities of OC cells.

Results: In this research, we demonstrated that TAM-derived exosomes facilitated OC cell proliferation and suppressed OC cell apoptosis. Then, qRT-PCR results indicated that miR-589-3p were markedly elevated after co-culture of TAM-derived exosomes with OC cells. In addition, we discovered that miR-589-3p was bound to BCL-2-like protein 13 (BCL2L13), which was confirmed through luciferase assay and RIP assay. Furthermore, functional analysis displayed that TAM-derived exosomes treated with miR-589-3p inhibitor attenuated the promotion of OC cell progression by exosomes.

Conclusion: TAM-derived exosomal miR-589-3p enhanced OC progression through BCL2L13, which offers a novel for OC therapy.

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来源期刊
Journal of Ovarian Research
Journal of Ovarian Research REPRODUCTIVE BIOLOGY-
CiteScore
6.20
自引率
2.50%
发文量
125
审稿时长
>12 weeks
期刊介绍: Journal of Ovarian Research is an open access, peer reviewed, online journal that aims to provide a forum for high-quality basic and clinical research on ovarian function, abnormalities, and cancer. The journal focuses on research that provides new insights into ovarian functions as well as prevention and treatment of diseases afflicting the organ. Topical areas include, but are not restricted to: Ovary development, hormone secretion and regulation Follicle growth and ovulation Infertility and Polycystic ovarian syndrome Regulation of pituitary and other biological functions by ovarian hormones Ovarian cancer, its prevention, diagnosis and treatment Drug development and screening Role of stem cells in ovary development and function.
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