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Tumor-derived exosomes encapsulating miR-34a promote apoptosis and inhibit migration and tumor progression of colorectal cancer cells under in vitro condition

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Abstract

Background

MicroRNA (miR)-34a, as a master tumor suppressor in colorectal cancer (CRC), could regulate multiple genes participating in tumor proliferation, invasion, immune evasion, and inflammation-induced progression. Exosomes, as novel nano-carriers, were found to be capable of shuttling crucial mediators to various cells. Since the conventional CRC therapeutics currently are a matter of debate, implication of microRNAs in malignancy remedies have been addressed illustrating promising outlooks.

Objectives

In this study, we aimed to investigate the delivery of miR-34a to CRC cell line CT-26 by encapsulating into tumor-derived exosomes (TEXs), in order to evaluate the anti-proliferative and progressive effects of the novel nano-carrier complex under in vitro condition.

Methods

Exosomes were purified from the starved CT-26 cells and then enriched by miR-34a using the calcium chloride (Cacl2) modified solution. Following the detection of miR-34a expression in the enriched TEXs, the viability of CT-26 cells treated by multiplicity concentrations of either TEXs or TEX-miR-34a was examined. Moreover, the apoptosis rate of the cells was evaluated, and the migration of CT-26 cells subjected to both TEX-miR-34a and TEX was also measured. Thereafter, the expressions of miR-34a target genes, as IL-6R, STAT3, PD-L1, and VEGF-A, which play roles in tumor progression, were determined in the treated CT-26 cells.

Results

The viability of CT-26 cells was harnessed following the treatment with TEX-miR-34a and the apoptosis levels of the cells were also observed to be enhanced dose-dependently. TEX-miR-34a was able to diminish the migration rate of the TEX-miR-34a treated cells and the expressions of IL-6R, STAT3, PD-L1, and VEGF-A were significantly restricted. Moreover, TEXs alone increased the apoptosis rate of tumor cells and repressed the proliferation and migration of these cells which were boosted by enrichment of TEXs with miR-34a.

Conclusion

Exosomes isolated from the starved CT-26 cells were found to have a potential to deliver miR-34a into tumor cells properly with high functionality maintenance for miR-34a in case of regulating genes related to tumor progression and TEXs which showed no positive effect favoring cancer cells, presumably act as a favorable adjuvant in the CRC therapy.

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Data availability

All data achieved during the current study are available by the corresponding author on reasonable request.

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Acknowledgments

The authors would like to acknowledge Zuhair Mohammad Hassan for his deliberate advice as well as Mahsa Hajivalili and Ardeshir Abbasi for their kind technical assistance in this project.

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Funding

This work was financially supported by Tarbiat Modares University and the National Animal Modeling Network and In vivo Research, Council for Development of Stem Cell Sciences and Technologies, Vice -Presidency for Science and Technology .grant number 52D/5147.

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MH, DA and ME planned the study, MH, DA and KB designed the methodology, MH and KB conducted the study. MH collected and analyzed the data. MH wrote the report. ME and DA were supervisors, KB was an adviser.

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Correspondence to Davar Amani or Masoumeh Ebtekar.

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Hosseini, M., Baghaei, K., Amani, D. et al. Tumor-derived exosomes encapsulating miR-34a promote apoptosis and inhibit migration and tumor progression of colorectal cancer cells under in vitro condition. DARU J Pharm Sci 29, 267–278 (2021). https://doi.org/10.1007/s40199-021-00400-0

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