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Article Open Access

M2‑TAM‑derived exosomal miR‑491‑3p modulates UBE2D3 and promotes the proliferation, migration and invasion of lung cancer cells

  • Authors:
    • Qiang Zhang
    • Yungang Sun
    • Yu Zhuang
    • Shiwei Xu
    • Mengxu Yao
    • Siyang Jiao
    • Qi Wang
    • Feng Shao
    • Xiaoying Zhang
  • View Affiliations / Copyright

    Affiliations: Department of Thoracic Surgery, Nanjing Chest Hospital, Nanjing, Jiangsu 210029, P.R. China, Wuxi School of Medicine, Jiangnan University, Wuxi 214000, P.R. China, The Third Affiliated Hospital of Soochow University, Tianning, Changzhou, Jiangsu 215004, P.R. China
    Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 159
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    Published online on: July 17, 2026
       https://doi.org/10.3892/or.2026.9164
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Abstract

M2 tumor‑associated macrophages (M2‑TAMs) have been reported to promote tumor growth through exosome‑dependent mechanisms. However, the exact role of exosomes derived from M2‑TAMs (M2‑TAM‑Exos) in lung cancer progression remains unclear. In The present study, M2‑like macrophages (IL‑4/IL‑13‑polarized THP‑1‑derived macrophages) were shown to release exosomes that lung cancer cells effectively internalized. These exosomes markedly enhanced the proliferation, migration, and invasion of lung cancer cells, thereby promoting malignancy. Further analyses revealed that M2‑like macrophage‑derived exosomes contain high levels of microRNA (miR)‑491‑3p. In vitro and in vivo experiments confirmed miR‑491‑3p as an oncogenic miR, while its inhibition markedly reduced cancer cell aggressiveness. Additional experiments demonstrated that miR‑491‑3p suppressed UBE2D3 expression after entering lung cancer cells. Collectively, these findings suggest a model in which M2‑like macrophages deliver miR‑491‑3p via exosomes to downregulate UBE2D3, facilitating lung cancer progression.
View Figures

Figure 1

M2-TAMs are markedly increased in
lung cancer tissues. (A) Representative images of CD68
immunohistochemical staining in lung cancer and adjacent normal
tissues. Scale bar, 50 µm. (B) Representative images of CD68
immunohistochemical staining in non-metastatic and metastatic lung
cancer tissues. Scale bar, 50 µm. (C) Statistical comparison of
CD68-positive areas between lung cancer tissues and adjacent normal
tissues. (D) Statistical analysis of CD68-positive areas between
non-metastatic and metastatic lung cancer tissues. (E) ROC curve
analysis for CD68 in lung cancer tissues. (F) mRNA expression
levels of M1 macrophage markers (IRF5 and TNF-α) in lung cancer
tissues compared with adjacent normal tissues. (G) mRNA expression
levels of M2 macrophage markers (ARG1 and IRF4) in lung cancer and
adjacent normal tissues. (H) mRNA expression levels of M2
macrophage markers (ARG1 and IRF4) in non-metastatic and metastatic
lung cancer tissues. Data are presented as mean ± SD.
***P<0.001. M2-TAMs, M2 tumor-associated macrophages; ROC,
receiver operating characteristic.

Figure 2

M2-like macrophages promote
proliferation, migration, and invasion of lung cancer cells. (A)
Flow cytometric analysis of CD206 expression in M0 and M2-like
macrophages. (B) ARG1 and IRF4 gene expression in M0 and M2-like
macrophages. (C) Proliferation curves of A549 and H460 lung cancer
cells treated with or without M2-CM. (D) Colony formation assays of
A549 and H460 lung cancer cells treated with or without M2-CM. (E)
Migration assays (24 h) of A549 and H460 cells treated with or
without M2-CM. Scale bar, 100 µm. (F) Invasion assays (24 h) of
A549 and H460 cells treated with or without M2-CM. Scale bar, 100
µm. (G) Quantitative analysis of cell invasion. (H) Quantitative
analysis of cell migration. (I) Subcutaneous tumor implantation of
A549 cells with or without M2-CM treatment. (J) Tumor volume
changes in mice implanted with A549 cells with or without M2-CM
treatment. (K) Tumor weight of A549 ×enografts with or without
M2-CM treatment. Data are presented as mean ± SD. ***P<0.001.
CM, conditioned medium.

Figure 3

M2-like macrophages facilitate
proliferation, migration, and invasion of lung cancer cells via
exosomes. (A) TEM images showing exosome morphology. Scale bar, 100
nm. (B) NTA analysis of exosome diameter distribution. (C) Western
blotting of CD81, CD63, TSG101 and GM130 in exosomes and whole-cell
lysates. (D) Proliferation curves for A549 and H460 cells treated
with CM, M2-CM, or M2-CM plus GW4869. (E) Colony formation assays
for A549 and H460 cells treated with control medium, M2-CM, or
M2-CM plus GW4869. (F) Migration assays of A549 and H460 cells
treated with CM, M2-CM, or M2-CM plus GW4869. Scale bar, 100 µm.
(G) Invasion assays of A549 and H460 cells treated with CM, M2-CM,
or M2-CM plus GW4869. Scale bar, 100 µm. (H) Representative images
of subcutaneous tumors derived from A549 cells treated with CM,
M2-CM, or M2-CM plus GW4869. (I) Tumor weights in mice implanted
with A549 and H460 cells treated with CM, M2-CM, or M2-CM plus
GW4869. (J) Tumor volume changes in mice implanted with A549 and
H460 cells treated with control medium, M2-CM, or M2-CM plus
GW4869. Data are presented as mean ± SD. **P<0.01,
***P<0.001. TEM, transmission electron microscopy; NTA,
nanoparticle tracking analysis; CM, conditioned medium.

Figure 4

M2-like macrophages release exosomes
enriched in miR-491-3p affecting lung cancer cells. (A) Heatmap of
differentially expressed miRNAs regulating lung cancer progression
from the GEO database. (B) Expression levels of miR-20a, miR-21,
miR-454, miR-491-3p, miR-140-3p, and miR-221 between control
(medium-only) and exosomes derived from M2-like macrophages. (C)
Expression levels of miR-20a, miR-21, miR-454, miR-491-3p,
miR-140-3p and miR-221 in HBE135-E6E7, A549 and H460 cells. (D)
Expression levels of precursor and mature miR-491-3p in A549 and
H460 cells. (E) Schematic diagram illustrating that Dil-labeled
exosomes from M2-like macrophages are internalized by A549 and H460
lung cancer cells. Scale bar, 10 µm. Data are presented as mean ±
SD. *P<0.05, **P<0.01, ***P<0.001. miR/miRNAs, microRNAs;
GEO, Gene Expression Omnibus.

Figure 5

Inhibition of miR-491-3p attenuates
lung cancer progression. (A) Verification of reduced miR-491-3p
expression after transfection with antagomiR-491-3p. (B)
Proliferation curves of A549 and H460 cells in antagomiR-NC and
antagomiR-491-3p groups. (C) Colony formation and quantitative
analysis of A549 and H460 cells in antagomiR-NC and
antagomiR-491-3p groups. (D) Migration assays of A549 and H460
cells in antagomiR-NC and antagomiR-491-3p groups. Scale bar, 100
µm. (E) Invasion assays of A549 and H460 cells in antagomiR-NC and
antagomiR-491-3p groups. Scale bar, 100 µm. (F) Quantitative
analysis of migration and invasion in A549 and H460 cells. (G)
Subcutaneous tumor formation, tumor weight and tumor volume in mice
implanted with A549 cells from antagomiR-NC and antagomiR-491-3p
groups. Data are presented as mean ± SD. **P<0.01,
***P<0.001. miR/miRNAs, microRNAs; NC, negative control.

Figure 6

miR-491-3p promotes lung cancer
progression by inhibiting UBE2D3 expression. (A) mRNA expression of
UBE2D3, AZI2, and PTPRM in HBE135-E6E7, A549, and H460 cells. (B)
Western blotting of UBE2D3 protein expression in HBE135-E6E7, A549,
and H460 cells. (C) mRNA levels of UBE2D3 in 68 paired clinical
lung cancer tissues and adjacent normal tissues. (D) mRNA
expression levels of UBE2D3 in A549 and H460 cells treated with
antagomiR-NC or antagomiR-491-3p. (E) ROC curve analysis for UBE2D3
expression in lung cancer tissues. (F) Colony formation and
quantitative analysis of A549 and H460 cells treated with control,
antagomiR-491-3p, and antagomiR-491-3p+sh-UBE2D3. (G) Migration
assays and quantitative analysis in A549 and H460 cells treated
with control, antagomiR-491-3p, and antagomiR-491-3p+sh-UBE2D3.
Scale bar, 100 µm. (H) Invasion assays and quantitative analysis in
A549 and H460 cells treated with control, antagomiR-491-3p and
antagomiR-491-3p+sh-UBE2D3. Scale bar, 100 µm. Data are presented
as mean ± SD. *P<0.05, **P<0.01, ***P<0.001, ##P<0.01,
###P<0.001. miR/miRNAs, microRNAs; NC, negative control; ROC,
receiver operating characteristic; sh, short hairpin.
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Copy and paste a formatted citation
Spandidos Publications style
Zhang Q, Sun Y, Zhuang Y, Xu S, Yao M, Jiao S, Wang Q, Shao F and Zhang X: M2‑TAM‑derived exosomal miR‑491‑3p modulates UBE2D3 and promotes the proliferation, migration and invasion of lung cancer cells. Oncol Rep 56: 159, 2026.
APA
Zhang, Q., Sun, Y., Zhuang, Y., Xu, S., Yao, M., Jiao, S. ... Zhang, X. (2026). M2‑TAM‑derived exosomal miR‑491‑3p modulates UBE2D3 and promotes the proliferation, migration and invasion of lung cancer cells. Oncology Reports, 56, 159. https://doi.org/10.3892/or.2026.9164
MLA
Zhang, Q., Sun, Y., Zhuang, Y., Xu, S., Yao, M., Jiao, S., Wang, Q., Shao, F., Zhang, X."M2‑TAM‑derived exosomal miR‑491‑3p modulates UBE2D3 and promotes the proliferation, migration and invasion of lung cancer cells". Oncology Reports 56.3 (2026): 159.
Chicago
Zhang, Q., Sun, Y., Zhuang, Y., Xu, S., Yao, M., Jiao, S., Wang, Q., Shao, F., Zhang, X."M2‑TAM‑derived exosomal miR‑491‑3p modulates UBE2D3 and promotes the proliferation, migration and invasion of lung cancer cells". Oncology Reports 56, no. 3 (2026): 159. https://doi.org/10.3892/or.2026.9164
Copy and paste a formatted citation
x
Spandidos Publications style
Zhang Q, Sun Y, Zhuang Y, Xu S, Yao M, Jiao S, Wang Q, Shao F and Zhang X: M2‑TAM‑derived exosomal miR‑491‑3p modulates UBE2D3 and promotes the proliferation, migration and invasion of lung cancer cells. Oncol Rep 56: 159, 2026.
APA
Zhang, Q., Sun, Y., Zhuang, Y., Xu, S., Yao, M., Jiao, S. ... Zhang, X. (2026). M2‑TAM‑derived exosomal miR‑491‑3p modulates UBE2D3 and promotes the proliferation, migration and invasion of lung cancer cells. Oncology Reports, 56, 159. https://doi.org/10.3892/or.2026.9164
MLA
Zhang, Q., Sun, Y., Zhuang, Y., Xu, S., Yao, M., Jiao, S., Wang, Q., Shao, F., Zhang, X."M2‑TAM‑derived exosomal miR‑491‑3p modulates UBE2D3 and promotes the proliferation, migration and invasion of lung cancer cells". Oncology Reports 56.3 (2026): 159.
Chicago
Zhang, Q., Sun, Y., Zhuang, Y., Xu, S., Yao, M., Jiao, S., Wang, Q., Shao, F., Zhang, X."M2‑TAM‑derived exosomal miR‑491‑3p modulates UBE2D3 and promotes the proliferation, migration and invasion of lung cancer cells". Oncology Reports 56, no. 3 (2026): 159. https://doi.org/10.3892/or.2026.9164
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