M2 macrophage-secreted exosomes promote metastasis and increase vascular permeability in hepatocellular carcinoma

Yiwei Lu; Guoyong Han; Yao Zhang; Long Zhang; Zhi Li; Qingyuan Wang; Zhiqiang Chen; Xuehao Wang; Jindao Wu

doi:10.1186/s12964-022-00872-w

M2 macrophage-secreted exosomes promote metastasis and increase vascular permeability in hepatocellular carcinoma

Cell Commun Signal. 2023 Oct 30;21(1):299. doi: 10.1186/s12964-022-00872-w.

Authors

Yiwei Lu^#^{1

2}, Guoyong Han^#^{1

2}, Yao Zhang^{1

2}, Long Zhang^{1

2}, Zhi Li³, Qingyuan Wang^{1

2}, Zhiqiang Chen^{1

2}, Xuehao Wang^{4

5

6}, Jindao Wu^#^{7

8

9}

Affiliations

¹ Department of Hepatobiliary Surgery, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, 210029, China.
² Key Laboratory of Liver Transplantation, Chinese Academy of Medical Sciences, NHC Key Laboratory of Living Donor Liver Transplantation,, Nanjing Medical University Nanjing, Nanjing, Jiangsu Province, China.
³ Department of General Surgery, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
⁴ Department of Hepatobiliary Surgery, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, 210029, China. wangxh@njmu.edu.cn.
⁵ Key Laboratory of Liver Transplantation, Chinese Academy of Medical Sciences, NHC Key Laboratory of Living Donor Liver Transplantation,, Nanjing Medical University Nanjing, Nanjing, Jiangsu Province, China. wangxh@njmu.edu.cn.
⁶ State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, China. wangxh@njmu.edu.cn.
⁷ Department of Hepatobiliary Surgery, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, 210029, China. wujindao@njmu.edu.cn.
⁸ Key Laboratory of Liver Transplantation, Chinese Academy of Medical Sciences, NHC Key Laboratory of Living Donor Liver Transplantation,, Nanjing Medical University Nanjing, Nanjing, Jiangsu Province, China. wujindao@njmu.edu.cn.
⁹ State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, China. wujindao@njmu.edu.cn.

^# Contributed equally.

Abstract

Background: Metastasis is a key feature of malignant tumors and significantly contributes to their high mortality, particularly in hepatocellular carcinoma (HCC). Therefore, it is imperative to explore the mechanism of tumor metastasis. Recently, tumor-associated macrophages (TAMs) have been demonstrated to promote tumor progression, while TAM-derived molecules involved in HCC metastasis warrant further investigation.

Methods: THP-1 was treated with IL-4 (Interleukin-4) and IL-13 (Interleukin-13) for M2 polarized macrophages. Exosomes derived from M2 macrophages were characterized. Then, HCC cells or human umbilical vein endothelial cells (HUVECs) were co-cultured with M2 macrophages or treated with M2 macrophage-secreted exosomes. Next, Transwell®, Scratch assay, tube formation, and endothelial permeability assays were performed. Moreover, RT-PCR, western blotting, immunofluorescence, and ELISA were used to assess mRNA and protein expression levels. Finally, the miRNA expression profiles of exosomes derived from M2 and M0 macrophages were analyzed.

Results: M2 macrophage infiltration was correlated with metastasis and a poor prognosis in HCC patients. M2-derived exosomes were absorbed by HCC and HUVEC cells and promoted the epithelial-mesenchymal transition (EMT), vascular permeability, and angiogenesis. Notably, MiR-23a-3p levels were significantly higher in M2-derived exosomes and hnRNPA1 mediated miR-23a-3p packaging into exosomes. Phosphatase and tensin homolog (PTEN) and tight junction protein 1 (TJP1) were the targets of miR-23a-3p, as confirmed by luciferase reporter assays. Lastly, HCC cells co-cultured with M2-derived exosomes secreted more GM-CSF, VEGF, G-CSF, MCP-1, and IL-4, which in turn further recruited M2 macrophages.

Conclusions: Our findings suggest that M2 macrophage-derived miR-23a-3p enhances HCC metastasis by promoting EMT and angiogenesis, as well as increasing vascular permeability. Video Abstract.

Keywords: Angiogenesis; EMT; Exosomes; M2 macrophages; Vascular permeability; miR-23a-3p.

Publication types

Video-Audio Media
Research Support, Non-U.S. Gov't

MeSH terms

Capillary Permeability
Carcinoma, Hepatocellular* / pathology
Cell Line, Tumor
Endothelial Cells / metabolism
Exosomes* / metabolism
Humans
Interleukin-4
Liver Neoplasms* / pathology
Macrophages / metabolism
MicroRNAs* / genetics
MicroRNAs* / metabolism

Substances

MicroRNAs
Interleukin-4