肿瘤相关巨噬细胞(TAMs)在肿瘤发展中起着重要作用。越来越多的研究表明，miR-210可能促进肿瘤恶性程度的进展，但它在原发性肝细胞癌(HCC)中的致癌作用是否通过对M2型巨噬细胞的作用尚未得到考察。以酯酸丙酯酯(PMA)和IL-4、IL-13诱导THP-1单核细胞分化为M2型极化的巨噬细胞。使用miR-210 mimics或miR-210 inhibitors转染M2型巨噬细胞。流式细胞术用于识别巨噬细胞相关标记物和细胞凋亡水平。通过qRT-PCR和Western blot检测M2型巨噬细胞的自噬水平、PI3K/AKT/mTOR信号通路相关的mRNAs和蛋白质表达。将HepG2和MHCC-97H HCC细胞系培养于M2细胞培养液中以探究M2型巨噬细胞源miR-210对HCC细胞增殖、迁移、侵袭和凋亡的影响。qRT-PCR显示M2型巨噬细胞中miR-210的表达增加。转染miR-210 mimics的M2型巨噬细胞中，自噬相关基因和蛋白质表达增强，而细胞凋亡相关蛋白质降低。MDC染色和透射电子显微镜观察到M2型巨噬细胞中miR-210 mimics组中MDC标记的小囊泡和自噬体的积累。在miR-210 mimics组中，M2型巨噬细胞中PI3K/AKT/mTOR信号通路的表达降低。与对照组相比，与转染miR-210 mimics的M2型巨噬细胞共培养的HCC细胞显示出增强的增殖和侵袭能力，而凋亡水平降低。此外，促进或抑制自噬可以增强或消除上述观察到的生物学效应。miR-210可以通过PI3K/AKT/mTOR信号通路促进M2型巨噬细胞的自噬。M2型巨噬细胞源miR-210通过自噬促进HCC的恶性进展，提示巨噬细胞自噬可能作为HCC的一个新的治疗靶点，而针对miR-210可能会重置M2型巨噬细胞在HCC上的作用。版权所有©2023 The Authors。由Elsevier Inc.发表。保留所有权利。

Tumor-associated macrophages (TAMs) play an important role in tumor development. Increasing research suggests that miR-210 may promote the progression of tumor virulence, but whether its pro-carcinogenic effect in primary hepatocellular carcinoma (HCC) is via an action on M2 macrophages has not been examined.Differentiation of THP-1 monocytes into M2-polarized macrophages was induced with phorbol myristate acetate (PMA) and IL-4, IL-13. M2 macrophages were transfected with miR-210 mimics or miR-210 inhibitors. Flow cytometry was used to identify macrophage-related markers and apoptosis levels. The autophagy level of M2 macrophages, expression of PI3K/AKT/mTOR signaling pathway-related mRNAs and protein were detected by qRT-PCR and Western blot. HepG2 and MHCC-97H HCC cell lines were cultured with M2 macrophages conditioned medium to explore the effects of M2 macrophage-derived miR-210 on the proliferation, migration, invasion and apoptosis of HCC cells.qRT-PCR showed increased expression of miR-210 in M2 macrophages. Autophagy-related gene and protein expression was enhanced in M2 macrophages transfected with miR-210 mimics, while apoptosis-related proteins were decreased. MDC staining and transmission electron microscopy observed the accumulation of MDC-labeled vesicles and autophagosomes in M2 macrophages in the miR-210 mimic group. The expression of PI3K/AKT/mTOR signaling pathway in M2 macrophages was reduced in miR-210 mimic group. HCC cells co-cultured with M2 macrophages transfected with miR-210 mimics exhibited enhanced proliferation and invasive ability as compared to the control group, while apoptosis levels were reduced. Moreover, promoting or inhibiting autophagy could enhance or abolish the above observed biological effects, respectively.miR-210 can promote autophagy of M2 macrophages via PI3K/AKT/mTOR signaling pathway. M2 macrophage-derived miR-210 promotes the malignant progression of HCC via autophagy, suggesting that macrophage autophagy may serve as a new therapeutic target for HCC, and targeting miR-210 may reset the effect of M2 macrophages on HCC.Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.