ZEB1 potentiates chemoresistance in breast cancer stem cells by evading apoptosis

Abstract

Chemoresistance renders a challenge to the clinics to treat breast cancer patients. Current treatment strategies are effective in mitigating tumor growth but remain largely ineffective against cancer-initiating cells or breast Cancer Stem Cells (CSCs). Epithelial-to-mesenchymal-transition (EMT) regulates breast CSC physiology. Zinc finger E-box binding homeobox 1 (ZEB1) is a key EMT-transcription factor that regulates breast CSC - differentiation and metastasis. However, its potential role in modulating tumor chemoresistance has not yet been fully understood. In-silico analysis revealed a higher ZEB1 expression in breast cancer patients that leads to decreased overall and relapse-free survival. We generated sorted breast CSC with stable ZEB1 overexpression (CD24^-/CD44⁺GFP-ZEB1) and/or silencing (CD24^-/CD44⁺ZEB1 shRNA) as well as breast cancer cells with stable ZEB1 overexpression (CD24⁺GFP-ZEB1) and/or silencing (CD24⁺ZEB1 shRNA). An increased colony-forming efficiency and doxorubicin accumulation correlated with decreased promoter activity and expression profile of ABCC1 drug-efflux ABC transporter in CD24^-/CD44⁺GFP-ZEB1. Additionally, CD24^-/CD44⁺GFP-ZEB1 demonstrated doxorubicin-induced higher anti-apoptotic and lower pro-apoptotic protein expressions in the mitochondrial and cytosolic fractions. Chemoresistant CD24^-/CD44⁺GFP-ZEB1 cells depicted 1000-fold higher IC-50 values of doxorubicin and decreased activation of JNK-p38 stress kinase molecular signaling-dependent mammosphere forming efficiency to evade apoptosis. Thus, ZEB1 and its downstream effectors are plausible therapeutic targets for the mitigation of breast cancer chemoresistance in patients.

Keywords: Anti-apoptosis; Breast cancer stem cells; Chemoresistance; JNK-p38 signaling; Tumorigenesis; ZEB1.