K Kalinsky1,2, T Zheng3, H Hibshoosh4,5, X Du4, P Mundi6, J Yang6, S Refice6, S M Feldman4,7, B Taback4,7, E Connolly4,8, K D Crew6,4,9, M A Maurer6,4, D L Hershman6,4,9. 1. Department of Medicine, College of Physicians and Surgeons, Columbia University Medical Center, New York, USA. kk2693@cumc.columbia.edu. 2. Herbert Irving Comprehensive Cancer Center, Columbia University, 161 Fort Washington Avenue, 10th Floor, Room 1069, New York, USA. kk2693@cumc.columbia.edu. 3. Department of Statistics, Columbia University, New York, USA. 4. Herbert Irving Comprehensive Cancer Center, Columbia University, 161 Fort Washington Avenue, 10th Floor, Room 1069, New York, USA. 5. Department of Pathology and Cell Biology, College of Physicians and Surgeons, Columbia University Medical Center, New York, USA. 6. Department of Medicine, College of Physicians and Surgeons, Columbia University Medical Center, New York, USA. 7. Department of Surgery, College of Physicians and Surgeons, Columbia University Medical Center, New York, USA. 8. Department of Radiation Oncology, College of Physicians and Surgeons, Columbia University Medical Center, New York, USA. 9. Department of Epidemiology and Biostatistics, Mailman School of Public Health, Columbia University, New York, USA.
Abstract
PURPOSE: Reverse Phase Protein Array (RPPA) is a high-throughput antibody-based technique to assess cellular protein activity. The goal of this study was to assess protein marker changes by RPPA in tumor tissue from a pre-surgical metformin trial in women with operable breast cancer (BC). METHODS: In an open-label trial, metformin 1500-mg PO daily was administered prior to resection in 35 non-diabetic patients with stage 0-III BC, body mass index ≥25 kg/m2. For RPPA, formalin-fixed paraffin-embedded (FFPE) samples were probed with 160 antibodies. Paired and two-sample t-tests were performed (p ≤ 0.05). Multiple comparisons were adjusted for by fixing the false discovery rate at 25 %. We evaluated whether pre- and post-metformin changes of select markers by RPPA were identified by immunohistochemistry (IHC) in these samples. We also assessed for these changes by western blot in metformin-treated BC cell lines. RESULTS: After adjusting for multiple comparisons in the 32 tumors from metformin-treated patients vs. 34 untreated historical controls, 11 proteins were significantly different between cases vs. CONTROLS: increases in Raptor, C-Raf, Cyclin B1, Cyclin D1, TRFC, and Syk; and reductions in pMAPKpT202,Y204, JNKpT183,pT185, BadpS112, PKC.alphapS657, and SrcpY416. Cyclin D1 change after metformin by IHC was not observed. In cell lines, reductions in JNKpT183 and BadpS112 were seen, with no change in Cyclin D1 or Raptor. CONCLUSIONS: These results suggest that metformin modulates apoptosis/cell cycle, cell signaling, and invasion/motility. These findings should be assessed in larger metformin trials. If confirmed, associations between these changes and BC clinical outcome should be evaluated. CLINICALTRIALS. GOV IDENTIFIER: NCT00930579.
PURPOSE: Reverse Phase Protein Array (RPPA) is a high-throughput antibody-based technique to assess cellular protein activity. The goal of this study was to assess protein marker changes by RPPA in tumor tissue from a pre-surgical metformin trial in women with operable breast cancer (BC). METHODS: In an open-label trial, metformin 1500-mg PO daily was administered prior to resection in 35 non-diabeticpatients with stage 0-III BC, body mass index ≥25 kg/m2. For RPPA, formalin-fixed paraffin-embedded (FFPE) samples were probed with 160 antibodies. Paired and two-sample t-tests were performed (p ≤ 0.05). Multiple comparisons were adjusted for by fixing the false discovery rate at 25 %. We evaluated whether pre- and post-metformin changes of select markers by RPPA were identified by immunohistochemistry (IHC) in these samples. We also assessed for these changes by western blot in metformin-treated BC cell lines. RESULTS: After adjusting for multiple comparisons in the 32 tumors from metformin-treated patients vs. 34 untreated historical controls, 11 proteins were significantly different between cases vs. CONTROLS: increases in Raptor, C-Raf, Cyclin B1, Cyclin D1, TRFC, and Syk; and reductions in pMAPKpT202,Y204, JNKpT183,pT185, BadpS112, PKC.alphapS657, and SrcpY416. Cyclin D1 change after metformin by IHC was not observed. In cell lines, reductions in JNKpT183 and BadpS112 were seen, with no change in Cyclin D1 or Raptor. CONCLUSIONS: These results suggest that metformin modulates apoptosis/cell cycle, cell signaling, and invasion/motility. These findings should be assessed in larger metformin trials. If confirmed, associations between these changes and BC clinical outcome should be evaluated. CLINICALTRIALS. GOV IDENTIFIER: NCT00930579.
Entities:
Keywords:
Breast Cancer; Cyclin D1; Immunohistochemistry; Metformin; Pre-surgical; Reverse Phase Protein Array
Authors: Kanwal P Raghav; Wenting Wang; Shuying Liu; Mariana Chavez-MacGregor; Xiaolong Meng; Gabriel N Hortobagyi; Gordon B Mills; Funda Meric-Bernstam; George R Blumenschein; Ana M Gonzalez-Angulo Journal: Clin Cancer Res Date: 2012-02-28 Impact factor: 12.531
Authors: Raoul Tibes; Yihua Qiu; Yiling Lu; Bryan Hennessy; Michael Andreeff; Gordon B Mills; Steven M Kornblau Journal: Mol Cancer Ther Date: 2006-10 Impact factor: 6.261
Authors: Mitch Dowsett; Steve R Ebbs; J Michael Dixon; Anthony Skene; Clive Griffith; Irene Boeddinghaus; Janine Salter; Simone Detre; Margaret Hills; Susan Ashley; Stephen Francis; Geraldine Walsh; Ian E Smith Journal: J Clin Oncol Date: 2005-03-14 Impact factor: 44.544
Authors: Bolin Liu; Zeying Fan; Susan M Edgerton; Xin-Sheng Deng; Irina N Alimova; Stuart E Lind; Ann D Thor Journal: Cell Cycle Date: 2009-07-21 Impact factor: 4.534
Authors: I Ben Sahra; K Laurent; A Loubat; S Giorgetti-Peraldi; P Colosetti; P Auberger; J F Tanti; Y Le Marchand-Brustel; F Bost Journal: Oncogene Date: 2008-01-21 Impact factor: 9.867
Authors: Sao Jiralerspong; Shana L Palla; Sharon H Giordano; Funda Meric-Bernstam; Cornelia Liedtke; Chad M Barnett; Limin Hsu; Mien-Chie Hung; Gabriel N Hortobagyi; Ana M Gonzalez-Angulo Journal: J Clin Oncol Date: 2009-06-01 Impact factor: 44.544
Authors: K Kalinsky; J A Sparano; X Zhong; E Andreopoulou; B Taback; L Wiechmann; S M Feldman; P Ananthakrishnan; A Ahmad; S Cremers; A N Sireci; J R Cross; D K Marks; P Mundi; E Connolly; K D Crew; M A Maurer; H Hibshoosh; S Lee; D L Hershman Journal: Clin Transl Oncol Date: 2018-05-07 Impact factor: 3.405
Authors: Maria Eduarda Azambuja Amaral; Laura Roesler Nery; Carlos Eduardo Leite; Walter Filgueira de Azevedo Junior; Maria Martha Campos Journal: Invest New Drugs Date: 2018-02-02 Impact factor: 3.850
Authors: Ting-Yuan David Cheng; Angela R Omilian; Song Yao; Weizhou Zhang; Susmita Datta; Wiam Bshara; Rochelle Payne Ondracek; Warren Davis; Song Liu; Chi-Chen Hong; Elisa V Bandera; Thaer Khoury; Christine B Ambrosone Journal: Breast Cancer Res Date: 2021-07-30 Impact factor: 6.466