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KCNQ1OT1 affects cell proliferation, invasion, and migration through a miR-34a / Notch3 axis in breast cancer

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Abstract

Background

Breast cancer (BC) accounts for a significant share of cancer-related deaths worldwide. Ongoing investigations have shown that long non-coding RNAs (lncRNAs) drive BC progression but their underlying mechanisms remain largely undescribed. LncRNA KCNQ1OT1 was previously identified in BC but its functional significance remained to be fully investigated.

Methods

KCNQ1OT1 and its downstream target genes were analyzed in breast cancer tissues and cell lines using methods including RT-qPCR, immunohistochemistry and Western blotting. The effects of KCNQ1OT1, miR-34a and Notch3 on BC cells were investigated using assays measuring proliferation (CCK-8, colony formation), apoptosis, and migration/invasion (scratch and Transwell assays). MS2-RIP and dual-luciferase reporter assays were used to study RNA interactions. Xenograft studies were employed to define the tumorigenic potential of KCNQ1OT1 in vivo.

Results

KCNQ1OT1 expression was up-regulated in BC tissues and high levels were associated with poorer prognosis. ShRNA inhibition of KCNQ1OT1 expression in BC cell lines retarded proliferation, migration and invasion in vitro and tumor growth in vivo. Up-regulation of KCNQ1OT1 was shown to inhibit miR-34a which was associated with blocking the inhibitory effect of miR-34a on BC cell proliferation, migration and invasion. Notch3 was found to be a downstream target of miR-34a with KCNQ1OT1 markedly inducing Notch3 expression in BC. Evidence for KCNQ1OT1/miR-34a/Notch3 axis was further established in clinical BC samples.

Conclusion

We identified a KCNQ1OT1/miR-34a/Notch3 axis which promotes BC progression through effects on cell proliferation and metastasis that was further associated with poor patient prognosis. These results propose targeting this axis as novel treatment approach for BC.

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Data Availability

All data generated or analyzed during this study are included in this published article.

Abbreviations

BC:

Breast cancer

KCNQ1OT1:

KCNQ1 overlap** transcript

ER:

Estrogen receptor

PR:

Progesterone receptor

HER-2:

Human epidermal growth factor receptor 2

KCNQ1OT1:

KCNQ1 overlap** transcript 1

LncRNA:

Long non-coding RNA

miRNA:

MicroRNA

ncRNA:

Non-coding RNA

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Acknowledgements

This work was supported by Wu Jie** Medical Foundation (No. 320.6750.2020-7-1) to MR. The authors would like to express their gratitude to EditSprings (https://www.editsprings.cn/) for the expert linguistic services provided.

Funding

This study was supported by Wu Jie** Medical Foundation (No. 320.6750.2020–7-1) to MR.

Author information

Author notes

  1. Zhiyao Ren, Yunfeng Xu and **n Wang contributed equally to this work

Authors and Affiliations

  1. Department of Breast Surgery, Department of General Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Ji** Avenue, Hefei, 230022, Anhui, PR China

    Zhiyao Ren, Yunfeng Xu & Min Ren

  2. Department of Plastic Surgery, The First Affiliated Hospital of Anhui Medical University, 218 Ji** Avenue, Hefei, 230022, Anhui, PR China

    **n Wang

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  1. Zhiyao Ren
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Contributions

ZYR and MR conceived and designed the research. ZYR, YFX and XW carried out experiments and analyzed the data. ZYR, YFX and XW wrote the main manuscript. All authors read and approved the manuscript and agree to be accountable for all aspects of the research in ensuring that the accuracy or integrity of any part of the work are appropriately investigated and resolved.

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Correspondence to Min Ren.

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The ethics approval and consent to participate was obtained from The First Affiliated Hospital of Anhui Medical University, Hefei, China.

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Ren, Z., Xu, Y., Wang, X. et al. KCNQ1OT1 affects cell proliferation, invasion, and migration through a miR-34a / Notch3 axis in breast cancer. Environ Sci Pollut Res 29, 28480–28494 (2022). https://doi.org/10.1007/s11356-021-18434-x

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