NRG1-dependent activation of HER3 induces primary resistance to trastuzumab in HER2-overexpressing breast cancer cells

Yang,Liuting; Li,Yingying; Shen,Enyun; Cao,Fengqi; Li,Li; Li,Xiaojin; Wang,Xuejiang; Kariminia,Seyed; Chang,Bingmei; Li,Hongzhong; Li,Qin

doi:10.3892/ijo.2017.4130

NRG1-dependent activation of HER3 induces primary resistance to trastuzumab in HER2-overexpressing breast cancer cells

Authors:
- Liuting Yang
- Yingying Li
- Enyun Shen
- Fengqi Cao
- Li Li
- Xiaojin Li
- Xuejiang Wang
- Seyed Kariminia
- Bingmei Chang
- Hongzhong Li
- Qin Li
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Affiliations: Department of Biochemistry and Molecular Biology, Basic Medical College, Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China, Department of Pathology and Pathophysiology, Basic Medical College, Capital Medical University, Beijing 100054, P.R. China, Beijing Cotimes Biotech Co., Ltd., Beijing 100176, P.R. China, Department of Oncology, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, P.R. China, Experimental Center, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, P.R. China, Molecular and Cellular Oncology, MD Anderson Cancer Center, Houston, TX 77030, USA, Chongqing Key Laboratory of Molecular Oncology and Epigenetics, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400416, P.R. China
Published online on: September 21, 2017 https://doi.org/10.3892/ijo.2017.4130
Pages: 1553-1562

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Abstract

This study was conducted to determine the role of neuregulin 1 (NRG1)-dependent human epidermal growth factor receptor 3 (HER3) activation in trastuzumab primary resistance, and to observe the inhibitory effect of HER3 monoclonal antibody on HER2-overexpressing breast cancer cells. BT474 cells (trastuzumab sensitive) and MDA-MB-453 cells (trastuzumab resistant) were first stimulated with NRG1 and then treated with either trastuzumab, HER3 antibody, or a combination of both. The expression of phospho human epidermal growth factor receptor 2 (p-HER2), phospho human epidermal growth factor receptor 3 (p-HER3), phospho protein kinase B (p-Akt) and phospho mitogen-activated protein kinase (p-MAPK) were detected by western blotting. Apoptosis was detected by flow cytometry. Cell viability was detected by MTT assay. Without NRG1 stimulation, trastuzumab treatment significantly downregulated the expression of p-HER2, increased early apoptosis, and decreased cell viability in BT474 cells. After NRG1 stimulation, the aforementioned effects weakened or disappeared in the trastuzumab treatment group, whereas in the HER3 antibody treatment group, there was significant downregulation in p-HER3 expression and increase in early apoptosis of BT474 cells. In MDA-MB-453 cells, the HER3 antibody significantly downregulated both p-HER2 and p-HER3 and promoted early apoptosis after NRG1 stimulation, however, trastuzumab hardly played a role. p-Akt and p-MAPK were also significantly downregulated by the HER3 antibody after NRG1 stimulation. The expressions of p-HER2, p-HER3, p-Akt and p-MAPK were all downregulated after HER3 gene silencing, compared to the control. NRG1-dependent activation of HER3 induces primary resistance to trastuzumab in HER2-overexpressing breast cancer cells. HER3 monoclonal antibody combined with trastuzumab may serve as a treatment choice for patients with primary resistance to trastuzumab.

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