The online version of this article (doi:10.1186/bcr2719) contains supplementary material, which is available to authorized users.
The authors declare that they have no competing interests.
IP coordinated sample collection, conducted immunohistochemical determination and scoring of markers Ki67, p-Akt and p-Erk1/2, acquired data on clinical-pathological parameters, conducted statistical analysis and interpretation of data for all markers and assisted in drafting the manuscript; FM selected eligible patients, collected samples in theatre and assisted in drafting the manuscript; MH sectioned tissue blocks and conducted immunohistochemical determination and scoring of markers ER, PgR and HER2; SD assisted in the immunohistochemical determination of phospho-markers; RA supervised the statistical analysis and assisted in drafting the manuscript; AN and PO supervised the histopathological processing of samples and contributed towards the histopathological discussions; JS provided training and supervision of the immunohistochemical tests; IS contributed to the design of the study and its interpretation; and MD conceived and designed the study, interpreted data and drafted the manuscript. All authors approved the final manuscript.
Very few studies have investigated whether the time elapsed between surgical resection and tissue fixation or the difference between core-cut and excision biopsies impact on immunohistochemically measured biomarkers, including phosphorylated proteins in primary breast cancer. The aim of this study was to characterise the differences in immunoreactivity of common biomarkers that may occur (1) as a result of tissue handling at surgery and (2) between core-cuts and resected tumours.
Core-cuts taken from surgical breast cancer specimens immediately after resection (sample A) and after routine X-ray of the excised tumour (sample B) were formalin-fixed and paraffin-embedded and compared with the routinely fixed resection specimen (sample C). The variation in immunohistochemical expression of Ki67, oestrogen receptor (ER), progesterone receptor (PgR), human epidermal growth factor 2 (HER2), p-Akt and p-Erk1/2 were investigated.
Twenty-one tissue sets with adequate tumour were available. Median time between collection of core-cuts A and B was 30 minutes (range, 20 to 80 minutes). None of the markers showed significant differences between samples A and B. Similarly, Ki67, ER, PgR and HER2 did not differ significantly between core-cuts and main resection specimen, although there was a trend for lower resection values for ER (P = 0.06). However, p-Akt and p-Erk1/2 were markedly lower in resections than core-cuts (median, 27 versus 101 and 69 versus 193, respectively; both P < 0.0001 [two-sided]). This difference was significantly greater in mastectomy than in lumpectomy specimens for p-Erk1/2 (P = 0.01).
The delay in fixation in core-cuts taken after postoperative X-ray of resection specimens has no significant impact on expression of Ki67, ER, PgR, HER2, p-Akt or p-Erk1/2. However, extreme loss of phospho-staining can occur during routine fixation of resection specimens. These differences are likely attributable to suboptimal fixation and may have major repercussions for clinical research involving these markers.
Dowsett M: Preoperative models to evaluate endocrine strategies for breast cancer. Clin Cancer Res. 2003, 9: 502s- PubMed
Wolff AC, Hammond ME, Schwartz JN, Hagerty KL, Allred DC, Cote RJ, Dowsett M, Fitzgibbons PL, Hanna WM, Langer A, McShane LS, Paik S, Pegram MD, Perez EA, Press MF, Rhodes A, Sturgeon C, Taube SE, Tubbs R, Vance GH, Vijver M, Wheeler TM, Hayes DF: American Society of Clinical Oncology/College of American Pathologists Guideline Recommendations for Human Epidermal Growth Factor Receptor 2 Testing in Breast Cancer. J Clin Oncol. 2007, 25: 118-145. 10.1200/JCO.2006.09.2775. CrossRefPubMed
Dowsett M, Bundred NJ, Decensi A, Sainsbury RC, Lu Y, Hills MJ, Cohen FJ, Veronesi P, O'Brien MER, Scott T, Muchmore DB: Effect of raloxifene on breast cancer cell Ki67 and apoptosis: a double-blind, placebo-controlled, randomized clinical trial in postmenopausal patients. Cancer Epidemiol Biomarkers Prev. 2001, 10: 961-966. PubMed
Dowsett M, Dixon JM, Horgan K, Salter J, Hills M, Harvey E: Antiproliferative effects of idoxifene in a placebo-controlled trial in primary human breast cancer. Clin Cancer Res. 2000, 6: 2260-2267. PubMed
Arnedos M, Nerurkar A, Osin P, A'Hern R, Smith IE, Dowsett M: Discordance between core needle biopsy (CBN) and excisional biopsy (EB) for estrogen receptor (ER), progesterone receptor (PgR) and HER2 status in early breast cancer (EBC). Ann Oncol. 2009, 20: 1948-1952. 10.1093/annonc/mdp234. CrossRefPubMed
Cuzick J, Dowsett M, Wale C, Salter J, Quinn E, Zabaglo L, Howell A, Buzdar A, Forbes J: Prognostic value of a combined ER, PgR, Ki67, HER2 immunohistochemical (IHC4) score and comparison with the GHI recurrence score: results from TransATAC. Cancer Res. 2009, 69 (Suppl): 503s-
Gutierrez MC, Detre S, Johnston S, Mohsin SK, Shou J, Allred DC, Schiff R, Osborne CK, Dowsett M: Molecular changes in tamoxifen-resistant breast cancer: relationship between estrogen receptor, HER-2, and p38 mitogen-activated protein kinase. J Clin Oncol. 2005, 23: 2469-2476. 10.1200/JCO.2005.01.172. CrossRefPubMed
Svensson S, Jirstrom K, Ryden L, Roos G, Emdin S, Ostrowski MC, Landberg G: ERK phosphorylation is linked to VEGFR2 expression and Ets-2 phosphorylation in breast cancer and is associated with tamoxifen treatment resistance and small tumours with good prognosis. Oncogene. 2005, 24: 4370-4379. 10.1038/sj.onc.1208626. CrossRefPubMed
Frogne T, Laenkholm AV, Lyng MB, Henriksen KL, Lykkesfeldt AE: Determination of HER2 phosphorylation at tyrosine 1221/1222 improves prediction of poor survival for breast cancer patients with hormone receptor-positive tumors. Breast Cancer Res. 2009, 11: R11-10.1186/bcr2230. CrossRefPubMedPubMedCentral
Modi S, DiGiovanna MP, Lu Z, Moskowitz C, Panageas KS, Van Poznak C, Hudis CA, Norton L, Tan L, Stern DF, Carter D, Seidman AD: Phosphorylated/activated HER2 as a marker of clinical resistance to single agent taxane chemotherapy for metastatic breast cancer. Cancer Invest. 2005, 23: 483-487. 10.1080/07357900500201301. CrossRefPubMed
Cappuzzo F, Magrini E, Ceresoli GL, Bartolini S, Rossi E, Ludovini V, Gregorc V, Ligorio C, Cancellieri A, Damiani S, Spreafico A, Paties CT, Lombardo L, Calandri C, Bellezza G, Tonato M, Crinò L: Akt phosphorylation and gefitinib efficacy in patients with advanced non-small-cell lung cancer. J Natl Cancer Inst. 2004, 96: 1133-1141. 10.1093/jnci/djh217. CrossRefPubMed
- Extreme loss of immunoreactive p-Akt and p-Erk1/2 during routine fixation of primary breast cancer
Isabel F Pinhel
Fiona A MacNeill
Margaret J Hills
Ian E Smith
- BioMed Central
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