Guodong Han1, Ni Qiu1, Kai Luo1, Hongling Liang1, Hongsheng Li1. 1. Department of Breast Surgery, Affiliated Cancer Hospital & Institute of Guangzhou Medical University, Guangzhou, Guangdong, P. R. China.
Abstract
BACKGROUND: microRNAs are involved in the control of cell growth and apoptosis; they also play an essential role in resistance towards trastuzumab, in breast cancer. The objective of this study was to identify differentially expressed microRNA(s) and explore its therapeutic role in treatment of the disease. METHODS: Real-time polymerase chain reaction (RT-PCR) was performed to identify the virtual microRNA (miRNA) involved in breast cancer cells resistant to trastuzumab. RT-PCR and Western blot analysis were carried out to study the effects of microRNA-141 (miR-141) on ERBB2, ERBB4 and AKT production. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenylterazolium bromide assay and flow cytometry analysis was carried out to examine the effect of miR-141 on cell proliferation and apoptosis via ERBB4. RESULT: According to RT-PCR results, only miR-141 and miR-375 among miR-141, miR-375, miR-16, miR-155, miR-217 and miR-205 were downregulated in trastuzumab-resistant cells. Trastuzumab-resistant cells displayed higher levels of ERBB4 and p-AKT as well as showing a higher growth rate and a lower apoptosis rate. Online software programs were used, which identified ERBB4 as a gene targeted by miR-141 with a highly conserved binding site for miR-141 located within the ERBB4 3'-untranslated region. In trastuzumab-resistant cells, miR-141 and shERBB4 reduced ERBB4 and p-AKT levels; ERBB2 and total AKT levels in miR-141 and shERBB4 groups showed no significant difference. Anti-miR-141 was upregulated ERBB4 and p-AKT levels in parental cell and had no obvious effect on ERBB2 and total AKT levels. Finally, miR-141 upregulated viability of the cells, which was restored by shERBB4, miR-141 and shERBB4 inhibited proliferation, and enhanced apoptosis of trastuzumab-resistant cells. miR-141 inhibitor caused an evident increase in proliferation and an obvious decrease in apoptosis of parental cells. CONCLUSION: Knockdown of miR-141 causes overexpression of ERBB4, which is involved in trastuzumab resistance in breast cancer cells. This study has implications that miR-141 as well as its target, ERBB4, as a potential target for treating trastuzumab-resistant breast cancers.
BACKGROUND: microRNAs are involved in the control of cell growth and apoptosis; they also play an essential role in resistance towards trastuzumab, in breast cancer. The objective of this study was to identify differentially expressed microRNA(s) and explore its therapeutic role in treatment of the disease. METHODS: Real-time polymerase chain reaction (RT-PCR) was performed to identify the virtual microRNA (miRNA) involved in breast cancer cells resistant to trastuzumab. RT-PCR and Western blot analysis were carried out to study the effects of microRNA-141 (miR-141) on ERBB2, ERBB4 and AKT production. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenylterazolium bromide assay and flow cytometry analysis was carried out to examine the effect of miR-141 on cell proliferation and apoptosis via ERBB4. RESULT: According to RT-PCR results, only miR-141 and miR-375 among miR-141, miR-375, miR-16, miR-155, miR-217 and miR-205 were downregulated in trastuzumab-resistant cells. Trastuzumab-resistant cells displayed higher levels of ERBB4 and p-AKT as well as showing a higher growth rate and a lower apoptosis rate. Online software programs were used, which identified ERBB4 as a gene targeted by miR-141 with a highly conserved binding site for miR-141 located within the ERBB4 3'-untranslated region. In trastuzumab-resistant cells, miR-141 and shERBB4 reduced ERBB4 and p-AKT levels; ERBB2 and total AKT levels in miR-141 and shERBB4 groups showed no significant difference. Anti-miR-141 was upregulated ERBB4 and p-AKT levels in parental cell and had no obvious effect on ERBB2 and total AKT levels. Finally, miR-141 upregulated viability of the cells, which was restored by shERBB4, miR-141 and shERBB4 inhibited proliferation, and enhanced apoptosis of trastuzumab-resistant cells. miR-141 inhibitor caused an evident increase in proliferation and an obvious decrease in apoptosis of parental cells. CONCLUSION: Knockdown of miR-141 causes overexpression of ERBB4, which is involved in trastuzumab resistance in breast cancer cells. This study has implications that miR-141 as well as its target, ERBB4, as a potential target for treating trastuzumab-resistant breast cancers.
Authors: Fernanda Cardoso da Silva; Angelo Borges de Melo Neto; Christina Aparecida Martins; Thaís Cunha de Sousa Cardoso; Matheus de Souza Gomes; Thaise Gonçalves de Araújo; Cristina Ribas Fürstenau Journal: Purinergic Signal Date: 2021-11-05 Impact factor: 3.765