MiR-548t-5p regulates pancreatic ductal adenocarcinoma metastasis through an IL-33-dependent crosstalk between cancer cells and M2 macrophages

ABSTRACT

IL-33 has been associated with pro- and anticancer functions in cancer. However, its role in pancreatic cancer metastasis remains unknown. This study aimed to explore the role of miR-548t-5p/IL-33 axis in the metastasis of pancreatic cancer. Luciferase activity assay, qRT-PCR, Western blot and ELISA were performed to prove whether IL-33 is the target of miR-548t-5p. In vivo metastasis assay and cellular transwell assay were performed to explore the role of miR-548t-5p/IL-33 axis in the invasion and metastasis of pancreatic cancer. Co-culture experiments and immunohistochemistry were performed to observe whether IL-33 affects cell invasion and metastasis dependent on the involvement of M2 macrophages. THP-1 cell induction experiment and flow cytometry were performed to explore the effect of IL-33 on macrophage polarization. CCK-8, colony formation, cell apoptosis, cell cycle, cell wound healing and transwell assay were performed to investigate the effect of IL-33 induced M2 macrophages on cell malignant biological behavior by coculturing pancreatic cancer cells with the conditioned medium (CM) from macrophages. We found that miR-548t-5p regulated the expression and secretion of IL-33 in pancreatic cancer cells by directly targeting IL-33 mRNA. IL-33 secreted by cancer cells promoted the recruitment and activation of macrophages to a M2-like phenotype. In turn, IL-33 induced M2 macrophages promoted the migration and invasion of cancer cells. Moreover, IL-33 affected pancreatic cancer cell invasion dependent on the involvement of M2 macrophages in the co-culture system. Thus, our study suggested that manipulation of this IL-33-dependent crosstalk has a therapeutic potential for the treatment of pancreatic cancer metastasis.

KEYWORDS:

• Pancreatic cancer
• IL-33
• miR-548t-5p
• tumor-associated macrophage
• metastasis

List of abbreviations

TAMs	=	tumor-associated macrophages
PMA	=	phorbol myristate acetate
MOI	=	multiplicity of infection
RT	=	reverse transcription
TMA	=	tissue microarray
IHC	=	immunohistochemistry
CM	=	conditioned medium
CCK-8	=	cell count kit-8

Disclosure statement

No potential conflict of interest was reported by the author(s).

Authors’ contributions

All authors contributed to the study conception and design. W. Y., G. W. and W. S drafted the article. Z. J. and Z. X. revised it critically for important intellectual content. W. Y., G. W., W. S, L. Y., Z. Z. and H. Y. contributed to acquisition of data, analysis and interpretation of data. All authors read and approved the final paper.

Availability of data and materials

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Ethics approval and consent to participate

This study was approved by the Ethics Committee of the First Affiliated Hospital of Nanjing Medical University and all patients provided written informed consent. All animal experiments were performed in accordance with animal protocols approved by the Nanjing Medical University.

Supplementary material

Supplemental data for this article can be accessed online at https://doi.org/10.1080/15384101.2024.2309026

Additional information

Funding

This work was supported by grants from the National Natural Science Foundation of China [81572337] (to Jing-Jing Zhang), the Innovation Capability Development Project of Jiangsu Province [No. BM2015004], and the Jiangsu Province Capability Improvement Project through Science, Technology and Education (Jiangsu Provincial Medical Key Discipline, ZDXK202222).