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A novel long non-coding RNA regulates the integrin, ITGA2 in breast cancer

  • Preclinical study
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Breast Cancer Research and Treatment Aims and scope Submit manuscript

Abstract

Purpose

ITGA2 encodes the integrin, α2 which mediates metastatic progression, and is a predictor of poor prognosis and chemoresistance in breast cancer. Decreased ITGA2 promoter methylation is implicated as a driver of increased gene expression in aggressive prostate and pancreatic tumours, however the contribution of altered methylation to ITGA2 expression changes in breast tumours has not been examined.

Methods

ITGA2 gene methylation and gene expression was examined in publicly available breast cancer datasets, and ITGA2 promoter methylation was mapped by targeted bisulphite sequencing analysis in breast tumour cell lines. The expression of a putative regulatory long noncoding RNA (lncRNA) was examined by qPCR and its’ functionality was investigated using gene knockdown (antisense oligonucleotides) and over expression in breast cancer cell lines.

Results

In breast tumours and breast cancer cell lines the ITGA2 promoter is largely unmethylated, with gene expression variable in tumour subtypes, irrespective of promoter methylation. A novel lncRNA (AC025180.1;ENSG00000249899), named herein I2ALR, was identified at the ITGA2 gene locus, and was variably expressed in breast tumours and breast cancer cell subtypes. I2LAR knockdown resulted in upregulation of ITGA2 gene expression, whilst over-expression of I2ALR resulted in downregulation of ITGA2 mRNA. Further, examination of two downstream targets of ITGA2 associated with breast tumor stemness and metastasis (CCND1 and ACLY), revealed concomitant gene expression changes in response to I2ALR modulation.

Conclusion

I2ALR represents a novel regulatory molecule targeting ITGA2 expression in breast tumours; a finding of significant and topical interest to the development of therapeutics targeting this integrin.

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Data availability

All data generated or analysed during this study are included in this published article and its supplementary information files. Publicly available data that support the findings of this study are openly available https://www.cbioportal.org/.

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Acknowledgements

The authors would like to convey their sincere thanks to participants of the Tasmanian Familial Prostate Cancer study who have generously consented to participate in this study and allow their stored pathology samples to be used for the purposes of this research. The authors would like to acknowledge Dr Kelsie Raspin provided cDNA samples for PrCa cell lines.

Funding

This study was supported by the Andree Greenwood Secondary Breast Cancer Fund; the Hobart Police Charity Trust; and the Royal Hobart Hospital Research Foundation. JLD is supported by a Select Foundation Cancer Research Fellowship, and previously supported by an Australian Research Council Future Fellowship.

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Author notes

  1. Adele F. Holloway and Joanne L. Dickinson have contributed equally to this work.

Authors and Affiliations

  1. Menzies Institute for Medical Research, College of Health and Medicine, University of Tasmania, Hobart, 7000, Australia

    Tristan Joseph Verhoeff & Joanne L. Dickinson

  2. Tasmanian School of Medicine, College of Health and Medicine, University of Tasmania, Hobart, 7000, Australia

    Adele F. Holloway

Authors
  1. Tristan Joseph Verhoeff
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Contributions

JLD and AFH conceived and designed the work, TJV performed laboratory experiments, analysed the data. TJV prepared the original draft of the work. TJV, JLD and AFH interpreted the data, and participated in revisions and editing of the manuscript.

Corresponding author

Correspondence to Joanne L. Dickinson.

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Verhoeff, T.J., Holloway, A.F. & Dickinson, J.L. A novel long non-coding RNA regulates the integrin, ITGA2 in breast cancer. Breast Cancer Res Treat 192, 89–100 (2022). https://doi.org/10.1007/s10549-021-06496-x

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