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01-09-2019 | Breast Cancer | Preclinical study

Triple-negative breast cancer cell line sensitivity to englerin A identifies a new, targetable subtype

Authors: Corena V. Grant, Chase M. Carver, Shayne D. Hastings, Karthik Ramachandran, Madesh Muniswamy, April L. Risinger, John A. Beutler, Susan L. Mooberry

Published in: Breast Cancer Research and Treatment | Issue 2/2019

Abstract

Purpose

Triple-negative breast cancers (TNBCs) represent a heterogeneous group of tumors. The lack of targeted therapies combined with the inherently aggressive nature of TNBCs results in a higher relapse rate and poorer overall survival. We evaluated the heterogeneity of TNBC cell lines for TRPC channel expression and sensitivity to cation-disrupting drugs.

Methods

The TRPC1/4/5 agonist englerin A was used to identify a group of TNBC cell lines sensitive to TRPC1/4/5 activation and intracellular cation disruption. Quantitative RT-PCR, the sulforhodamine B assay, pharmacological inhibition, and siRNA-mediated knockdown approaches were employed. Epifluorescence imaging was performed to measure intracellular Ca²⁺ and Na⁺ levels. Mitochondrial membrane potential changes were monitored by confocal imaging.

Results

BT-549 and Hs578T cells express high levels of TRPC4 and TRPC1/4, respectively, and are exquisitely, 2000- and 430-fold, more sensitive to englerin A than other TNBC cell lines. While englerin A caused a slow Na⁺ and nominal Ca²⁺ accumulation in Hs578T cells, it elicited rapid increases in cytosolic Ca²⁺ levels that triggered mitochondrial depolarization in BT-549 cells. Interestingly, BT-549 and Hs578T cells were also more sensitive to digoxin as compared to other TNBC cell lines. Collectively, these data reveal TRPC1/4 channels as potential biomarkers of TNBC cell lines with dysfunctional mechanisms of cation homeostasis and therefore sensitivity to cardiac glycosides.

Conclusions

The sensitivity of BT-549 and Hs578T cells to englerin A and digoxin suggests a subset of TNBCs are highly susceptible to cation disruption and encourages investigation of TRPC1 and TRPC4 as potential new biomarkers of sensitivity to cardiac glycosides.

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Title: Triple-negative breast cancer cell line sensitivity to englerin A identifies a new, targetable subtype
Authors: Corena V. Grant
Chase M. Carver
Shayne D. Hastings
Karthik Ramachandran
Madesh Muniswamy
April L. Risinger
John A. Beutler
Susan L. Mooberry
Publication date: 01-09-2019
Publisher: Springer US
Keywords: Breast Cancer
Cardiac Glycoside
Digoxin
Breast Cancer
Published in: Breast Cancer Research and Treatment / Issue 2/2019
Print ISSN: 0167-6806
Electronic ISSN: 1573-7217
DOI: https://doi.org/10.1007/s10549-019-05324-7

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Abstract

Purpose

Methods

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