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Cancer cell-derived exosomal LINC00313 induces M2 macrophage differentiation in non-small cell lung cancer

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Abstract

Purpose

Non-small cell lung cancer (NSCLC) is the major subtype of lung cancer, which is the leading cause of cancer death worldwide. Tumor-associated macrophages (TAMs) are one of the main non-tumor cells in the tumor microenvironment. Here, we investigated the effect of cancer cell-derived exosomal LINC00313 on the M2 macrophage differentiation in NSCLC and clarified its underlying mechanism.

Methods

Flow cytometry, Western blotting, ELISA and immunohistochemical staining were performed to identify the macrophage phenotype by detecting the expression of M2 markers. The expression levels of LINC00313 and miR-135a-3p were measured by qRT-PCR, and luciferase reporter assay was used to validate the binding of lncRNA to miRNA, and miRNA to the target gene STAT6. The mouse-xenograft models were established by subcutaneous injection of the NCl-H1299 cells with stable overexpression or knockdown of LINC00313. GW4869 was injected intra-tumorally after tumor implantation.

Results

It was found that the cancer cells promoted M2 macrophage differentiation by secreting exosomes. LINC00313 was overexpressed in H1299-derived exosomes, and its knockdown abolished the effect of H1299-induced M2 macrophage differentiation. LINC00313 sponged miR-135a-3p to increase the STAT6 expression, resulting in the M2 macrophage differentiation. LINC00313 promoted tumor progression and promoted the expression of M2 markers in isolated tumor macrophages. A novel regulatory mechanism of M2 macrophage differentiation in NSCLC was revealed. It was found that cancer cell-derived exosomal LINC00313 promoted M2 macrophage differentiation in NSCLC by up-regulating STAT6 as miR-135a-3p sponge.

Conclusions

This study provides a new mechanism and direction to prevent the M2 macrophage differentiation in NSCLC.

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Acknowledgements

We would like to thank all the researchers and subjects for their contributions.

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Author notes

  1. Wencui Kong and Lei Zhang contributed equally to this work.

Authors and Affiliations

  1. Department of Pulmonary and Critical Care Medicine, 900TH Hospital of Joint Logistics Support Force, Fuzhou, 350025, Fujian, China

    Wencui Kong, Lei Zhang, Ying Chen, Zongyang Yu & Zhongquan Zhao

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  1. Wencui Kong
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Contributions

WK conceived and designed the study, and drafted the manuscript. LZ and YC collected, analyzed and interpreted the experimental data. ZY and ZZ revised the manuscript for important intellectual content. All authors read and approved the final manuscript.

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Correspondence to Zhongquan Zhao.

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The authors declare that they have no conflict of interest.

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The study was approved by Ethical Committee of 900TH Hospital of Joint Logistics Support Force and conducted in accordance with the ethical standards.

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Figure S1. LINC01116 was related to the prognosis of NSCLC and up-regulated in exosomes from NSCLC cells.

(A) TCGA database showed that the five lncRNAs (LINC01116, LINC00968, LINC00313, lncRNA C1orf132 and lncRNA TMPO-AS1) were related to the prognosis of NSCLC. (B) qRT-PCR showed an increased LINC00313 level in NSCLC cells (H1299, A549, H1975) and MRC-5. n=3, data were described by mean ± SEM. * (*p<0.05; **p<0.01; ***p<0.001) was considered different from control group. (JPG 2198 KB)

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Kong, W., Zhang, L., Chen, Y. et al. Cancer cell-derived exosomal LINC00313 induces M2 macrophage differentiation in non-small cell lung cancer. Clin Transl Oncol 24, 2395–2408 (2022). https://doi.org/10.1007/s12094-022-02907-7

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