Livin participates in resistance to trastuzumab therapy for breast cancer through ERK1/2 and AKT pathways and promotes EMT-like phenotype

Abstract

Trastuzumab resistance has emerged as a major issue in anti-human epidermal growth factor receptor-2 (HER2) therapy for breast cancers. The cell lines maintain overexpression of HER2. Upon treatment with trastuzumab, R-SKBR3 and R-BT474 cell lines displayed an increased growth rate and invasiveness, accompanied by activation of the ERK1/2 and AKT signaling pathways, and also a parental EMT-like transition (epithelial–mesenchymal transition) was promoted, with increases in N-cadherin, vimentin, and fibronectin and a decrease in E-cadherin. A further investigation found that livin played a key role in the development of trastuzumab resistance. Knockdown of the expression of livin by livin-shRNA3 in R-SKBR3 and R-BT474 cells decreased ERK1/2 and AKT, resensitized the resistant cells to the therapeutic activities of trastuzumab by inducing growth arrest, inhibition of proliferation, and G1-S cell cycle checking in the presence of the antibody, and they also exhibited an EMT-like transition (epithelial–mesenchymal transition), with a decrease in N-cadherin and an increase in E-cadherin, and the cell invasiveness was inhibited in response to the downregulation of livin. Conversely, SKBR3 and BT474 cells that had been stably transfected with pcDNA3.1-livin underwent promotion of an EMT-like transition and displayed a significant decrease in E-cadherin and increases in N-cadherin, vimentin, and fibronectin, and ectopic expression of livin in HER2-overexpressing breast cancer cells conferred resistance to trastuzumab. In vivo, the administration of livin AS (antisense oligonucleotides) restored sensitivity to trastuzumab in resistant breast cancer xenografts via the ERK1/2 and AKT signaling pathways. Patients with livin-overexpressing breast cancers exhibited significantly poorer responses to trastuzumab-based therapy than those with normal livin levels. In summary, our data suggest that the upregulation of livin activates the ERK1/2 and AKT signaling pathways and promotes an EMT-like transition. This could be an important mechanism that leads to trastuzumab resistance in HER2-overexpressing breast cancer cells.