The Effect of Freezing on the Immunoprofile of Breast Carcinoma Cells

Abstract

Background: Intraoperative frozen section procedure may be required in some operations performed for breast masses. Aims: We investigated the effect of frozen section procedure on the immunoprofile of breast carcinoma cells. Study Design: Cross-sectional study. Methods: A total of 53 breast carcinoma cases evaluated with intraoperative frozen sections were included in this study. Immunohistochemically, oestrogen (ER), progesterone (PR) and HER2 primary antibodies were evaluated in both frozen and non-frozen sections of each tumour sample. Results: No difference was found between the frozen and non-frozen sections in 33 cases in terms of staining rate and intensity of ER, PR and HER2. A decrease was found in the ER, PR and HER2 staining rate (in 10 cases, 9 cases and 6 cases, respectively). Likewise, a decrease was detected in ER, PR and HER2 staining intensity in 6 patients for each. Although there was no staining in frozen sections, immunopositivity was observed in 3 non-frozen sections for ER, in 2 for PR and in 5 for HER2. Statistically, a significant difference was found between the frozen and non-frozen sections in terms of staining rate and intensity for each of the three markers. Conclusion: In this study, the frozen section procedure in breast carcinoma had a negative effect on the immunoprofile. However, considering the importance of hormone receptor status in the treatment, these results should be supported with larger series

Keywords

• Breast carcinoma, frozen examination, HER2, immunohistochemistry, oestrogen, progesterone

Kaynakça

1. 1. Jemal A, Siegel R, Xu J, Ward E. Cancer statistics, 2010. CA Cancer J Clin 2010;60:277-300. [CrossRef]
2. 2. Early Breast Cancer Trialists’ Collaborative Group. Effects of chemotherapy and hormonal therapy for early breast cancer on recurrence and 15-year survival: an overview of the randomised trials. Lancet 2005;365:1687-717.[CrossRef]
3. 3. Burstein HJ,Harris JR, Morrow M. Malignant tumors of the breast. In: DeVita VT, Lawrence TS, Rosenberg SA, eds. DeVita, Hellman, and Rosenberg’s Cancer Principles & practice of oncology. 9th ed. Philadelphia: Lippincott Williams & Wilkins; 2011.p.1401-46.
4. 4. Adamczyk A1, Niemiec J, Ambicka A, Małecki K, Wysocki WH, Mituś J, et al. Expression of ER/PR/HER2, basal markers and adhesion molecules in primary breast cancer and in lymph nodes metastases: a comparative immunohistochemical analysis. Pol J Pathol 2012;63:228-34.
5. 5. Gown AM. Current issues in ER and HER2 testing by IHC in breast cancer. Mod Pathol 2008;21(Suppl 2):S8-15.[CrossRef]
6. 6. Leake R, Barnes D, Pinder S, Ellis I, Anderson L, Anderson T, et al. Immunohistochemical detection of steroid receptors in breast cancer: a working protocol. UK Receptor Group, UK NEQAS, The Scottish Breast Cancer Pathology Group, and The Receptor and Biomarker Study Group of the EORTC. J Clin Pathol 2000;53:634-5.[CrossRef]
7. 7. Rhodes A, Jasani B, Balaton AJ, Miller KD. Immunohistochemical demonstration of estrogen and progesterone receptors: correlation of standards achieved on in house tumours with that achieved on external quality assessment material in over 150 laboratories from 26 countries. J Clin Pathol 2000;53:292-301.[CrossRef]
8. 8. Rhodes A, Jasani B, Balaton AJ, Barnes DM, Anderson E, Bobrow LG, et al. Study of interlaboratory reliability and reproducibility of estrogen and progesterone receptor assays in Europe: documentation of poor reliability and identification of insufficient microwave antigen retrieval time as a major contributory element of unreliable assays. Am J Clin Pathol 2001;115:44-58. [CrossRef]

9. 9. Oyama T, Ishikawa Y, Hayashi M, Arihiro K, Horiguchi J. The effects of fixation, processing and evaluation criteria on immunohistochemical detection of hormone receptors in breast cancer. Breast Cancer 2007;14:182-8.[CrossRef]
10. 10. Grizzle WE. Special symposium: Fixation and tissue processing models. Biotech Histochem 2009;84:185-93. [CrossRef]
11. 11. Fitzgibbons PL, Murphy DA, Hammond ME, Allred DC, Valenstein PN. Recommendations for validating estrogen and progesterone receptor immunohistochemistry assays. Arch Pathol Lab Med 2010;134:930-5.
12. 12. Goldstein NS, Ferkowicz M, Odish E, Mani A, Hastah F. Minimum formalin fixation time for consistent estrogen receptor immunohistochemical staining of invasive breast carcinoma. Am J Clin Pathol 2003;120:86-92. [CrossRef]
13. 13. Apple S, Pucci R, Lowe AC, Shintaku I, Shapourifar-Tehrani S, Moatamed N. The effect of delay in fixation, different fixatives, and duration of fixation in estrogen and progesterone receptor results in breast carcinoma. Am J Clin Pathol 2011;135:592-8. [CrossRef]
14. 14. Khoury T, Sait S, Hwang H, Chandrasekhar R, Wilding G, Tan D, et al. Delay to formalin fixation effect on breast biomarkers. Mod Pathol 2009;22:1457-67. [CrossRef]
15. 15. De Marzo AM, Fedor HH, Gage WR, Rubin MA. Inadequate formalin fixation decreases reliability of p27 immunohistochemical staining: probing optimal fixation time using highdensity tissue microarrays. Hum Pathol 2002;33:756-60.[CrossRef]
16. 16. Grizzle WE, Stockard CR, Billings PE. The effects of tissue processing variables other than fixation on histochemical staining and immunohistochemical detection of antigens. J Histotechnol 2001;24:213-9. [CrossRef]
17. 17. Taylor CR, Shi SR. Antigen retrieval: call for a return to first principles. Appl Immunohistochem Mol Morphol 2000;8:173-4.[CrossRef]
18. 18. Werner M, Chott A, Fabiano A, Battifora H. Effect of formalin tissue fixation and processing on immunohistochemistry. Am J Surg Pathol 2000;24:1016-9.[CrossRef]
19. 19. Yamashita S, Okada Y. Application of heat-induced antigen retrieval to aldehyde-fixed fresh frozen sections. J Histochem Cytochem 2005;53:1421-32. [CrossRef]
20. 20. Shi SR, Liu C, Pootrakul L, Tang L, Young A, Chen R, Cote RJ, et al. Evaluation of the value of frozen tissue section used as “gold standard” for immunohistochemistry. Am J Clin Pathol 2008;129:358-66. [CrossRef]
21. 21. Eltoum I, Fredenburgh J, Myers RB, Grizzle WE. Introduction to the theory and practice of fixation of tissues. J Histotechnol 2001;24:173-90. [CrossRef]
22. 22. Karimi-Busheri F, Zadorozhny V, Carrier E, Fakhrai H. Molecular integrity and global gene expression of breast and lung cancer stem cells under long-term storage and recovery. Cell Tissue Bank 2013;14:175-86. [CrossRef]
23. 23. Caboux E, Paciencia M, Durand G, Robinot N, Wozniak MB, GalateauSalle F, et al. Impact of delay to cryopreservation on RNA integrity and genome-wide expression profiles in resected tumor samples. PLoS One 2013;8:e79826. [CrossRef]