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High-content siRNA 3D co-cultures to identify myoepithelial cell-derived breast cancer suppressor proteins

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posted on 2021-07-13, 04:25 authored by HM Duivenvoorden, Natasha Brockwell, CJ Nowell, KJ Simpson, Belinda ParkerBelinda Parker
Understanding how cancer cells interact with the surrounding microenvironment early in breast cancer development can provide insight into the initiation and progression of invasive breast cancers. The myoepithelial cell layer surrounding breast ducts acts as a physical barrier in early breast cancer, preventing cancer cells from invading the surrounding stroma. Changes to the expression profile and properties of myoepithelial cells have been implicated in progression to invasive carcinoma. Identifying the molecular drivers of myoepithelial cell-mediated tumour suppression may offer new approaches to predict and block the earliest stages of cancer invasion. We employed a high-content approach to knock down 87 different genes using siRNA in an immortalised myoepithelial cell line, prior to co-culture with invasive breast cancer cells in 3D. Combined with high-content imaging and a customised analysis pipeline, this system was used to identify myoepithelial proteins that are necessary to control cancer cell invasion. This dataset has identified prospective myoepithelial suppressors of early breast cancer invasion which may be used by researchers to investigate their clinical validity and utility.


This work was supported by grant funding from the National Health and Medical Research Council (NHMRC) (BSP 1047748 and 1127754); fellowship support to BSP from Victoria Cancer Agency and HMD from Cancer Council Victoria. The authors would like to thank Jennii Luu, Karla Cowley and Iva Nikolic from the Victorian Centre for Functional Genomics for their assistance in this project. The Victorian Centre for Functional Genomics (K.J.S.) is funded by the Australian Cancer Research Foundation (ACRF), Phenomics Australia, through funding from the Australian Government's National Collaborative Research Infrastructure Strategy (NCRIS) program and the Peter MacCallum Cancer Centre Foundation. We thank Kornelia Polyak for providing the N1ME cell line.


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