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CRISPR interference and activation of the microRNA-3662-HBP1 axis control progression of triple-negative breast cancer

Oncogene volume 41, pages 268–279 (2022)Cite this article

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Abstract

MicroRNA-3662 (miR-3662) is minimally expressed in normal human tissues but is highly expressed in all types of cancers, including breast cancer. As determined with The Cancer Genome Atlas dataset, miR-3662 expression is higher in triple-negative breast cancers (TNBCs) and African American breast cancers than in other breast cancer types. However, the functional role of miR-3662 remains a topic of debate. Here, we found that inhibition or knockout of endogenous, mature miR-3662 in TNBC cells suppresses proliferation and migration in vitro and tumor growth and metastasis in vivo. Functional analysis revealed that, for TNBC cells, knockout of miR-3662 reduces the activation of Wnt/β-catenin signaling. Furthermore, using CRISPR-mediated miR-3662 activation and repression, dual-luciferase assays, and miRNA/mRNA immunoprecipitation assays, we established that HMG-box transcription factor 1 (HBP-1), a Wnt/β-catenin signaling inhibitor, is a target of miR-3662 and is most likely responsible for miR-3662-mediated TNBC cell proliferation. Our results suggest that miR-3662 has an oncogenic function in tumor progression and metastasis via an miR-3662-HBP1 axis, regulating the Wnt /β-catenin signaling pathway in TNBC cells. Since miR-3662 expression occurs a tumor-specific manner, it is a promising biomarker and therapeutic target for patients who have TNBCs with dysregulation of miR-3662, especially African Americans.

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Fig. 1: miR-3662 KO inhibited cell proliferation and migration of TNBC cells.
Fig. 2: miR-3662 KO reduced tumor growth and metastasis of TNBC xenografts in NSG mice.
Fig. 3: miR-3662 KO repressed the activation of Wnt/β-catenin signaling in MDA-MB-231 cells.
Fig. 4: Identification of HBP1 as a target gene of miR-3662 in MDA-MB-231 cells.
Fig. 5: Validation of miR-3662-HBP1 axis-mediated regulation of the Wnt/β-catenin signaling pathway in BT-20 cells.
Fig. 6: miR-3662-mediated functional role and post-transcriptional regulation of HBP1.
Fig. 7: Regulation of the miR-3662-HBP1 axis and its effect on growth and the Wnt signaling pathway.

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Acknowledgements

We thank Dr. Donald Hill for editorial assistance in preparing this manuscript. This work was supported by grants from the National Cancer Institute (CA223077, CA238273 and CA242917 for R. Liu) and the Breast Cancer Research Foundation of Alabama (L. Wang). Results are based, in part, upon data generated by TCGA Research Network: http://cancergenome.nih.gov/.

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Authors and Affiliations

  1. Department of Genetics, University of Alabama at Birmingham, Birmingham, AL, USA

    Baozhu Yi, Shuaibin Wang, Xinran Wang, Zhichao Liu, Chao Zhang, Ming Li, Song Gao, Lizhong Wang & Runhua Liu

  2. O’Neal Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, AL, USA

    Shi Wei, Sejong Bae, Erica Stringer-Reasor, Lizhong Wang & Runhua Liu

  3. Department of Pathology, University of Alabama at Birmingham, Birmingham, AL, USA

    Shi Wei

  4. Department of Medicine, University of Alabama at Birmingham, Birmingham, AL, USA

    Sejong Bae & Erica Stringer-Reasor

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Contributions

Conception, design, and financial support: LW, RL. Development of methodology: BY, LW, RL. Acquisition of data (bench/animal works, acquired data, etc.): BY, SW, XW, ZL, CZ, ML, SG. Analysis and interpretation of data (e.g., statistical analysis, biostatistics, computational analysis): SW, SB, ES, LW, RL. Writing, review, and/or revision of the manuscript: BY, LW, RL. Pathology analysis: SW. Study supervision: LW, RL.

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Correspondence to Lizhong Wang or Runhua Liu.

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Yi, B., Wang, S., Wang, X. et al. CRISPR interference and activation of the microRNA-3662-HBP1 axis control progression of triple-negative breast cancer. Oncogene 41, 268–279 (2022). https://doi.org/10.1038/s41388-021-02089-6

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