MicroRNA-495/TGF-β/FOXC1 axis regulates multidrug resistance in metaplastic breast cancer cells

Uttom Kumar; Yunhui Hu; Nahal Masrour; Marcos Castellanos-Uribe; Alison Harrod; Sean T. May; Simak Ali; Valerie Speirs; R. Charles Coombes; Ernesto Yagüe

doi:10.1016/j.bcp.2021.114692

MicroRNA-495/TGF-β/FOXC1 axis regulates multidrug resistance in metaplastic breast cancer cells

Uttom Kumar, Yunhui Hu, Nahal Masrour, Marcos Castellanos-Uribe, Alison Harrod, Sean T. May, Simak Ali, Valerie Speirs, R. Charles Coombes, Ernesto Yagüe^* (Corresponding Author)

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

14 Citations (Scopus)

Abstract

Triple-negative metaplastic breast carcinoma (MBC) poses a significant treatment challenge due to lack of targeted therapies and chemotherapy resistance. We isolated a novel MBC cell line, BAS, which showed a molecular and phenotypic profile different from the only other metaplastic cell model, HS578T cells. To gain insight behind chemotherapeutic resistance, we generated doxorubicin (HS-DOX, BAS-DOX) and paclitaxel (HS-TX, BAS-TX) resistant derivatives of both cell lines. Drug sensitivity assays indicated a truly multidrug resistant (MDR) phenotype. Both BAS-DOX and BAS-TX showed up-regulation of FOXC1 and its experimental down-regulation re-sensitized cells to doxorubicin and paclitaxel. Experimental modulation of FOXC1 expression in MCF-7 and MDA-MB-231 cells corroborated its role in MDR. Genome-wide expression analyses identified gene expression signatures characterized by up-regulation of TGFB2, which encodes cytokine TGF-β2, in both BAS-DOX and BAS-TX cells. Pharmacological inhibition of the TGF-β pathway with galunisertib led to down-regulation of FOXC1 and increase in drug sensitivity in both BAS-DOX and BAS-TX cells. MicroRNA (miR) expression analyses identified high endogenous miR-495-3p levels in BAS cells that were downregulated in both BAS MDR cells. Transient expression of miR-495-3p mimic in BAS-DOX and BAS-TX cells caused downregulation of TGFB2 and FOXC1 and re-sensitized cells to doxorubicin and paclitaxel, whereas miR-495-3p inhibition in BAS cells led to increase in resistance to both drugs and up-regulation of TGFB2 and FOXC1. Together, these data suggest interplay between miR-495-3p, TGF-β2 and FOXC1 regulating MDR in MBC and open the exploration of novel therapeutic strategies.

Original language	English
Article number	114692
Number of pages	15
Journal	Biochemical Pharmacology
Volume	192
Early online date	21 Jul 2021
DOIs	https://doi.org/10.1016/j.bcp.2021.114692
Publication status	Published - 31 Oct 2021

Bibliographical note

Acknowledgements
We thank preliminary work by Deborah Holliday who generated the BAS cells from tissue provided by pathologist Andrew Hanby and Breast Cancer Now for funding that initial work. This work was funded by a Commonwealth Scholarship Commission bursary to Uttom Kumar. The National Institute for Health Research Imperial Biomedical Research Centre, the Imperial Experimental Cancer Medicine Centre and the Cancer Research UK Imperial Centre at Imperial College London provided infrastructure support. The authors wish to acknowledge the roles of the Breast Cancer Now Tissue Bank in collecting and making available samples, and the patients who have generously donated their tissues to be used in the generation of this publication.

Keywords

FOXC1
Galunisertib
Metaplastic breast cancer
miR-495-3p
Multidrug resistance
TGF-β

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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@article{907eb25e52a54f48a5f142849f689a18,

title = "MicroRNA-495/TGF-β/FOXC1 axis regulates multidrug resistance in metaplastic breast cancer cells",

abstract = "Triple-negative metaplastic breast carcinoma (MBC) poses a significant treatment challenge due to lack of targeted therapies and chemotherapy resistance. We isolated a novel MBC cell line, BAS, which showed a molecular and phenotypic profile different from the only other metaplastic cell model, HS578T cells. To gain insight behind chemotherapeutic resistance, we generated doxorubicin (HS-DOX, BAS-DOX) and paclitaxel (HS-TX, BAS-TX) resistant derivatives of both cell lines. Drug sensitivity assays indicated a truly multidrug resistant (MDR) phenotype. Both BAS-DOX and BAS-TX showed up-regulation of FOXC1 and its experimental down-regulation re-sensitized cells to doxorubicin and paclitaxel. Experimental modulation of FOXC1 expression in MCF-7 and MDA-MB-231 cells corroborated its role in MDR. Genome-wide expression analyses identified gene expression signatures characterized by up-regulation of TGFB2, which encodes cytokine TGF-β2, in both BAS-DOX and BAS-TX cells. Pharmacological inhibition of the TGF-β pathway with galunisertib led to down-regulation of FOXC1 and increase in drug sensitivity in both BAS-DOX and BAS-TX cells. MicroRNA (miR) expression analyses identified high endogenous miR-495-3p levels in BAS cells that were downregulated in both BAS MDR cells. Transient expression of miR-495-3p mimic in BAS-DOX and BAS-TX cells caused downregulation of TGFB2 and FOXC1 and re-sensitized cells to doxorubicin and paclitaxel, whereas miR-495-3p inhibition in BAS cells led to increase in resistance to both drugs and up-regulation of TGFB2 and FOXC1. Together, these data suggest interplay between miR-495-3p, TGF-β2 and FOXC1 regulating MDR in MBC and open the exploration of novel therapeutic strategies.",

keywords = "FOXC1, Galunisertib, Metaplastic breast cancer, miR-495-3p, Multidrug resistance, TGF-β",

author = "Uttom Kumar and Yunhui Hu and Nahal Masrour and Marcos Castellanos-Uribe and Alison Harrod and May, {Sean T.} and Simak Ali and Valerie Speirs and Coombes, {R. Charles} and Ernesto Yag{\"u}e",

note = "Acknowledgements We thank preliminary work by Deborah Holliday who generated the BAS cells from tissue provided by pathologist Andrew Hanby and Breast Cancer Now for funding that initial work. This work was funded by a Commonwealth Scholarship Commission bursary to Uttom Kumar. The National Institute for Health Research Imperial Biomedical Research Centre, the Imperial Experimental Cancer Medicine Centre and the Cancer Research UK Imperial Centre at Imperial College London provided infrastructure support. The authors wish to acknowledge the roles of the Breast Cancer Now Tissue Bank in collecting and making available samples, and the patients who have generously donated their tissues to be used in the generation of this publication.",

year = "2021",

month = oct,

day = "31",

doi = "10.1016/j.bcp.2021.114692",

language = "English",

volume = "192",

journal = "Biochemical Pharmacology",

issn = "0006-2952",

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TY - JOUR

T1 - MicroRNA-495/TGF-β/FOXC1 axis regulates multidrug resistance in metaplastic breast cancer cells

AU - Kumar, Uttom

AU - Hu, Yunhui

AU - Masrour, Nahal

AU - Castellanos-Uribe, Marcos

AU - Harrod, Alison

AU - May, Sean T.

AU - Ali, Simak

AU - Speirs, Valerie

AU - Coombes, R. Charles

AU - Yagüe, Ernesto

N1 - Acknowledgements We thank preliminary work by Deborah Holliday who generated the BAS cells from tissue provided by pathologist Andrew Hanby and Breast Cancer Now for funding that initial work. This work was funded by a Commonwealth Scholarship Commission bursary to Uttom Kumar. The National Institute for Health Research Imperial Biomedical Research Centre, the Imperial Experimental Cancer Medicine Centre and the Cancer Research UK Imperial Centre at Imperial College London provided infrastructure support. The authors wish to acknowledge the roles of the Breast Cancer Now Tissue Bank in collecting and making available samples, and the patients who have generously donated their tissues to be used in the generation of this publication.

PY - 2021/10/31

Y1 - 2021/10/31

N2 - Triple-negative metaplastic breast carcinoma (MBC) poses a significant treatment challenge due to lack of targeted therapies and chemotherapy resistance. We isolated a novel MBC cell line, BAS, which showed a molecular and phenotypic profile different from the only other metaplastic cell model, HS578T cells. To gain insight behind chemotherapeutic resistance, we generated doxorubicin (HS-DOX, BAS-DOX) and paclitaxel (HS-TX, BAS-TX) resistant derivatives of both cell lines. Drug sensitivity assays indicated a truly multidrug resistant (MDR) phenotype. Both BAS-DOX and BAS-TX showed up-regulation of FOXC1 and its experimental down-regulation re-sensitized cells to doxorubicin and paclitaxel. Experimental modulation of FOXC1 expression in MCF-7 and MDA-MB-231 cells corroborated its role in MDR. Genome-wide expression analyses identified gene expression signatures characterized by up-regulation of TGFB2, which encodes cytokine TGF-β2, in both BAS-DOX and BAS-TX cells. Pharmacological inhibition of the TGF-β pathway with galunisertib led to down-regulation of FOXC1 and increase in drug sensitivity in both BAS-DOX and BAS-TX cells. MicroRNA (miR) expression analyses identified high endogenous miR-495-3p levels in BAS cells that were downregulated in both BAS MDR cells. Transient expression of miR-495-3p mimic in BAS-DOX and BAS-TX cells caused downregulation of TGFB2 and FOXC1 and re-sensitized cells to doxorubicin and paclitaxel, whereas miR-495-3p inhibition in BAS cells led to increase in resistance to both drugs and up-regulation of TGFB2 and FOXC1. Together, these data suggest interplay between miR-495-3p, TGF-β2 and FOXC1 regulating MDR in MBC and open the exploration of novel therapeutic strategies.

AB - Triple-negative metaplastic breast carcinoma (MBC) poses a significant treatment challenge due to lack of targeted therapies and chemotherapy resistance. We isolated a novel MBC cell line, BAS, which showed a molecular and phenotypic profile different from the only other metaplastic cell model, HS578T cells. To gain insight behind chemotherapeutic resistance, we generated doxorubicin (HS-DOX, BAS-DOX) and paclitaxel (HS-TX, BAS-TX) resistant derivatives of both cell lines. Drug sensitivity assays indicated a truly multidrug resistant (MDR) phenotype. Both BAS-DOX and BAS-TX showed up-regulation of FOXC1 and its experimental down-regulation re-sensitized cells to doxorubicin and paclitaxel. Experimental modulation of FOXC1 expression in MCF-7 and MDA-MB-231 cells corroborated its role in MDR. Genome-wide expression analyses identified gene expression signatures characterized by up-regulation of TGFB2, which encodes cytokine TGF-β2, in both BAS-DOX and BAS-TX cells. Pharmacological inhibition of the TGF-β pathway with galunisertib led to down-regulation of FOXC1 and increase in drug sensitivity in both BAS-DOX and BAS-TX cells. MicroRNA (miR) expression analyses identified high endogenous miR-495-3p levels in BAS cells that were downregulated in both BAS MDR cells. Transient expression of miR-495-3p mimic in BAS-DOX and BAS-TX cells caused downregulation of TGFB2 and FOXC1 and re-sensitized cells to doxorubicin and paclitaxel, whereas miR-495-3p inhibition in BAS cells led to increase in resistance to both drugs and up-regulation of TGFB2 and FOXC1. Together, these data suggest interplay between miR-495-3p, TGF-β2 and FOXC1 regulating MDR in MBC and open the exploration of novel therapeutic strategies.

KW - FOXC1

KW - Galunisertib

KW - Metaplastic breast cancer

KW - miR-495-3p

KW - Multidrug resistance

KW - TGF-β

U2 - 10.1016/j.bcp.2021.114692

DO - 10.1016/j.bcp.2021.114692

M3 - Article

C2 - 34298004

SN - 0006-2952

VL - 192

JO - Biochemical Pharmacology

JF - Biochemical Pharmacology

M1 - 114692

ER -

MicroRNA-495/TGF-β/FOXC1 axis regulates multidrug resistance in metaplastic breast cancer cells

Abstract

Bibliographical note

Keywords

UN SDGs

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