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Knockdown of SF3B1 inhibits cell proliferation, invasion and migration triggering apoptosis in breast cancer via aberrant splicing

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Abstract

Background

Splicing factor 3b subunit 1 (SF3B1) was frequently reported to be significantly mutated in breast cancer. However, the status of SF3B1 expression, its function and molecular consequence in breast cancer remained unreported.

Methods

Immunohistochemistry was used to assess SF3B1expression in 110 breast cancer samples. SF3B1 knock‑down in ZR-75-30 and MDA-MB-231 cells was performed by shRNA transfection. The expression of SF3B1 in cells was detected by quantitative real‑time PCR and western blot. Cell proliferation ability was determined by MTT and colony formation assay. Migration and invasion were determined by transwell assay. Flow cytometry was performed to investigate cell cycle and apoptosis. RNA-sequencing was performed to examine differentially expressed genes and affected alternative splicing events.

Results

SF3B1 is overexpressed in breast cancer tissues compared with normal tissues. Overexpression of SF3B1 is associated with lymph node metastasis. SF3B1 knockdown in MDA-MB-231 and ZR-75-30 breast cancer cells significantly induced the suppression of proliferation, migration, invasion and also enhancement of apoptosis. RNA-sequencing data revealed that 860 genes were significantly up-regulated and 776 genes were significantly down-regulated upon SF3B1 knockdown. Differentially expressed genes enriched in the signaling pathways including Ras signaling pathway; cytokine receptor interaction; tight junction; MAPK signaling pathway, Glycine, serine and threonine metabolism. Alternative splicing analysis revealed that exon skipping (SKIP) and cassette exons (MSKIP) were the most common molecular effect upon SF3B1 knockdown.

Conclusions

Our study suggests that SF3B1 may be an important molecular target for breast cancer treatment and provides a new clue for clinical treatment of breast cancer.

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Acknowledgements

This work was supported by funding from the National Natural Science Foundation of China (81773150 to LZ.), Natural Science Foundation of Shanxi Province (201801D121339 to JL.), the Program for the Outstanding Innovative Teams of Higher Learning Institutions of Shanxi (OIT 2017 to LZ.), the Doctoral Start up Research Fund of Shanxi Medical University (03201508 to LZ.).

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Author notes

  1. Ling Zhang, Xiaojuan Zhang and Haitao Zhang are equal first authors.

Authors and Affiliations

  1. Department of Pathology, Shanxi Medical University, Taiyuan, Shanxi, 030001, People’s Republic of China

    Ling Zhang, Xiaojuan Zhang & Yanghui Bi

  2. Shanxi Key Laboratory of Carcinogenesis and Translational Research of Esophageal Cancer, Shanxi Medical University, Taiyuan, 030001, Shanxi, People’s Republic of China

    Ling Zhang, Xiaojuan Zhang & Yanghui Bi

  3. Department of General Surgery, The First Hospital of Shanxi Medical University, Taiyuan, 030001, Shanxi, People’s Republic of China

    Haitao Zhang, Yanyan Zhang & Jing Liu

  4. Department of Forensic Medicine, Shanxi Medical University, Taiyuan, 030001, Shanxi, People’s Republic of China

    Feng Liu

  5. Department of Pathology, The First Hospital of Shanxi Medical University, Taiyuan, 030001, Shanxi, People’s Republic of China

    Caixia Cheng

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  1. Ling Zhang
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Correspondence to Jing Liu.

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The authors declare no conflict of interest.

Ethical approval

The present study was approved by the Ethics Committee of the Shanxi Medical University (approval No. 2018LL0115) and was conducted in accordance with the ethical standards. All procedures performed in studies involving human participants were in accordance the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration.

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Zhang, L., Zhang, X., Zhang, H. et al. Knockdown of SF3B1 inhibits cell proliferation, invasion and migration triggering apoptosis in breast cancer via aberrant splicing. Breast Cancer 27, 464–476 (2020). https://doi.org/10.1007/s12282-020-01045-8

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  • DOI: https://doi.org/10.1007/s12282-020-01045-8

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