Translational Therapeutics

miR-4711-5p regulates cancer stemness and cell cycle progression via KLF5, MDM2 and TFDP1 in colon cancer cells

Abstract

Background

It is important to establish cancer stem cell (CSC)-targeted therapies to eradicate cancer. As it is a CSC marker, we focused on Kruppel-like factor 5 (KLF5) in this study.

Methods

We searched for candidate microRNAs (miRNAs) that inhibited KLF5 expression by in silico analyses and screened them in colon cancer cell lines.

Results

We identified one promising miRNA, miR-4711-5p, that downregulated KLF5 expression by direct binding. This miRNA suppressed cell proliferation, migration and invasion ability, as well as stemness, including decreased stem cell marker expression, reactive oxygen species activity and sphere formation ability. MiR-4711-5p inhibited the growth of DLD-1 xenografts in nude mice with no adverse effects. We found that miR-4711-5p provoked G1 arrest, which could be attributed to direct binding of miR-4711-5p to TFDP1 (a heterodimeric partner of the E2F family). Our findings also suggested that direct binding of miR-4711-5p to MDM2 could upregulate wild-type p53, leading to strong induction of apoptosis. Finally, we found that miR-4711-5p had a potent tumour-suppressive effect compared with a putative anti-oncomiR, miR-34a, in tumour cell cultures derived from five patients with colorectal cancer.

Conclusions

Our data suggest that miR-4711-5p could be a promising target for CSC therapy.

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Fig. 1: Identification of the KLF5-targeting miRNA miR-4711-5p.
Fig. 2: In CRC cell lines, miR-4711-5p exerted various anti-tumour effects.
Fig. 3: MiR-4711-5p suppressed the stemness of DLD-1 and HCT116 cells.
Fig. 4: MiR-4711-5p inhibited the G1-to-S phase transition in the cell cycle.
Fig. 5: Anti-tumour effects and safety in vivo. Systemic administration of formulated sCA-miR-4711-5p inhibited tumour growth.
Fig. 6: Anti-tumour effects in CRC cell lines and patient-derived tumor cells.

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Acknowledgements

We acknowledge the core NGS facility of the Genome Information Research Centre of Osaka University for their support in IPA and data analysis.

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Authors

Contributions

Conception and design: Y.M., M.M., Y.D., H.Y. Development of methodology: Y.M., T.M., X.W., H.Y. Acquisition of data: A.I., T.M., X.W., Y.Y., T.H., H.H., Y.Q., J.W., N.M., H.T., N.H., C.M., H.Y. Analysis and interpretation of data (e.g. statistical analysis, biostatistics and computational analysis): Y.M., D.O., M.M., Y.D., H.Y. Writing, review and/or revision of the manuscript: Y.M., T.M., Y.Y., H.H., H.T., H.Y. Administrative, technical or material support: Y.M., T.M., C.M., M.M., Y.D., H.Y. Study supervision: Y.M., T.M., M.M., Y.D., H.Y.

Corresponding author

Correspondence to Hirofumi Yamamoto.

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Ethics approval and consent to participate

All animal experiments were performed in accordance with currently prescribed guidelines and followed a protocol approved by Osaka University. This study was performed in accordance with the Declaration of Helsinki. Written informed consent was obtained from all patients, and the study was approved by the Ethics Board of Osaka University Hospital.

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Informed consent was obtained from all patients for publication of this article.

Data availability

The expression levels of miR-4711-5p and other miRNAs referred to in this study (Supplementary Fig. S6) are available on the Tissue Atlas website (https://ccb-web.cs.uni-saarland.de/tissueatlas/). The expression levels of KLF5 referred to in this study (Supplementary Fig. S1) are available on the Human Protein Atlas website (https://www.proteinatlas.org/). All the other data of the study can be found with the corresponding author.

Competing interests

The authors declare no competing interests.

Funding information

This work was supported by a grant from Kagoshima Shinsangyo Sousei Investment Limited Partnership (its general partner is Kagoshima Development Co., Ltd).

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Morimoto, Y., Mizushima, T., Wu, X. et al. miR-4711-5p regulates cancer stemness and cell cycle progression via KLF5, MDM2 and TFDP1 in colon cancer cells. Br J Cancer 122, 1037–1049 (2020). https://doi.org/10.1038/s41416-020-0758-1

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