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. 2021 Apr 22;11:8735. doi: 10.1038/s41598-021-87698-7

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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BRCA1 knockdown represses breast cancer cell growth and lower BRCA1 expression correlates with longer relapse-free survival rate. (a) MCF7 cells stably expressing Tet-shRNA vector against BRCA1 (Tet-shBRCA1) or control vector (Tet-shNT) were cultured with or without 0.1 μg/ml DOX for 48 h. BRCA1 knockdown efficiency was confirmed by Western blotting. (b) Growth of MCF7 cells expressing the Tet-shNT, Tet-shBRCA1 #1, or #3 vectors was assessed using the IncuCyte live-cell imaging system in the presence or absence of 0.1 μg/ml DOX for 4 days. n = 4, means ± S.D. *p < 0.05 versus -DOX. (c) Comparison of colony formation ability between MCF7 cells expressing Tet-shNT and Tet-shBRCA1. Cells were seeded in 6-well plates and cultured for 3 weeks in the presence or absence of 0.1 μg/ml DOX. Visualized colonies are shown in the left panel. Summarized colony numbers are shown in the right panel. Bar = means ± S.D. n = 3, *p < 0.05 versus -DOX. (d) The growth of PINK1 knockout MCF7 cells (sgPINK1#1, #2) or control cells (sgNega3, 5) was assessed using the IncuCyte live-cell imaging system. n = 5, means ± S.D. *p < 0.05 versus sgNega5. (e) Relapse-free survival rates of breast cancer patients were compared between high and low BRCA1, Parkin, or PINK1 expression in patients. Data are shown as Kaplan–Meier plots; graphs were generated using an online Kaplan–Meier plotter.