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. 2018 Mar 2;8:3930. doi: 10.1038/s41598-018-22250-8

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© The Author(s) 2018

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Metformin induced caveolin-1 expression and promoted T-DM1 efficacy. Western blot analysis demonstrated that when BT-474 cells were treated with metformin and other chemotherapy drugs, caveolin-1 expression was induced (A); this started to decrease at 72 hours when treated with metformin (B). (C) In BT-474 cells pretreated with metformin for 24 hours and then treated with a combination of T-DM1 and metformin, the cell viability was significantly decreased. (D) SKBR3 cells treated with metformin for 24 hours, caveolin-1 expression was induced. (E) SKBR3 treated with metformin combined with T-DM1 or pretreated with metformin and followed by T-DM1 treatment with or without metformin combination showed decreased cell survival compared to metformin or T-DM1 treated group. MET/M: metformin; PTX: paclitaxel; CY: cyclophosphamide; 5-FU: 5-fluorouracil; TRA: trastuzumab; T: T-DM1. Statistical values with different letters were significantly different (P < 0.01). At least three independent experiments were performed for all quantifications. The western blots were derived under the same experimental conditions from the same cell lysates of each treatment group in (A,B and D); the original full-length western blot images are showed in Supplementary Figure S2.