Comparing Heat-Induced Antigen Retrieval Methods for ER Expression in Invasive Breast Carcinoma: Invasive Breast Carcinoma

Shela Akbar; Hina Khan; Abdul Qadir

doi:10.54393/pjhs.v4i10.1120

Authors

Shela Akbar Department of Histopathology, Combined Military Hospital, Peshawar, Pakistan
Hina Khan Department of Histopathology, Combined Military Hospital, Peshawar, Pakistan
Abdul Qadir Department of Histopathology, Combined Military Hospital, Peshawar, Pakistan

DOI:

https://doi.org/10.54393/pjhs.v4i10.1120

Keywords:

Invasive Breast Carcinoma, ER Expression, Antigen Retrieval Methods

Abstract

In the realm of cancer diagnosis and treatment, immuno-histochemical staining has become a pivotal tool for discerning molecular markers within tissue samples. Objective: To comprehensively compare different heat-induced antigen retrieval methods for evaluating estrogen receptor (ER) expression in female invasive breast carcinoma of no special type. Methods: A cross-sectional design was employed utilizing 250 formalin-fixed paraffin-embedded (FFPE) tissue samples from confirmed cases of invasive breast carcinoma. The study was conducted within Histopathology Department in CMH Peshawar. The study spanned from January 6, 2022 to June 30, 2023. Three heat-induced antigen retrieval methods were evaluated: conventional microwave-based retrieval, pressure cooking retrieval, and water bath heating. ER expression was assessed through immune-histo-chemical staining, and scores were analyzed using descriptive statistics and ANOVA. A post hoc Tukey's Honestly Significant Difference (HSD) test was conducted to identify which specific retrieval methods exhibited significant differences in ER expression scores. Results: The pressure-cooking retrieval method exhibited significantly higher mean ER expression scores (7.52) compared to the conventional microwave-based (6.87) and water bath heating methods (6.65). Post hoc analysis confirmed the substantial differences. Standard deviations and interquartile ranges were consistent with each method. Conclusions: The study underscores the pivotal role of retrieval techniques in accurate ER expression assessment in female invasive breast carcinoma of no special type. The pressure cooking method emerges as a potent enhancer of ER expression, advocating for standardized protocols to ensure consistent diagnostic outcomes and informed therapeutic decisions. Despite limitations, this research contributes to the refinement of patient care in the management of invasive breast carcinoma.

References

Kornaga EN, Klimowicz AC, Guggisberg N, Ogilvie T, Morris DG, Webster M, et al. A systematic comparison of three commercial estrogen receptor assays in a single clinical outcome breast cancer cohort. Modern Pathology. 2016 Aug; 29(8): 799-809. doi: 10.1038/modpathol.2016.74. DOI: https://doi.org/10.1038/modpathol.2016.74

Tripathy K, Mishra A, Singh AK, Panda PL, Mahapatra A, Lenka A. Immunocytochemistry using liquid-based cytology: a tool in hormone receptor analysis of breast cancer. Journal of Cytology. 2018 Oct; 35(4): 260. doi: 10.4103/JOC.JOC_23_18. DOI: https://doi.org/10.4103/JOC.JOC_23_18

Makoui MH, Mobini M, Khoshnoodi J, Judaki MA, Bahadori T, Zare HA, et al. Generation and Characterization of Novel Diagnostic Mouse Monoclonal Antibodies Against Human Estrogen Receptor Alpha and Progesterone Receptor. Asian Pacific Journal of Cancer Prevention: APJCP. 2022 Sep; 23(9): 2999. doi: 10.31557/APJCP.2022.23.9.2999. DOI: https://doi.org/10.31557/APJCP.2022.23.9.2999

Alwahaibi NY, Albadi HM, Almasrouri NM, Alsinawi SS, Aldairi N. Immunocytochemical evaluation of estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2 in breast cancer cell blocks and corresponding tissue blocks: A single institutional experience. Journal of Medical Sciences. 2018 Jul; 38(4): 160-4. doi: 10.4103/jmedsci.jmedsci_130_17. DOI: https://doi.org/10.4103/jmedsci.jmedsci_130_17

Geethamala K, Murthy VS, Vani BR, Rao MS, Thejaswini MU, Padmaja KP. Comparison of immunocytochemistry and immunohistochemistry on breast carcinoma: A boon or a bane? Journal of Laboratory Physicians. 2017 Jan; 9(01): 005-10. doi: 10.4103/0974-2727.187915. DOI: https://doi.org/10.4103/0974-2727.187915

Kadivar M and Aram F. Assessment of ki67 in breast cancer: A comparison between the eye-10 method, stepwise counting strategy, and international system of ki67 evaluation. Iranian Journal of Pathology. 2020 Dec; 15(1): 13. doi: 10.30699/ijp.2019.102290.2017. DOI: https://doi.org/10.30699/ijp.2019.102290.2017

Cassoobhoy M and Shaw G. WebMD March/April 2014. Healthy Start. [Last Cited: 19th Mar 2023]. Available at: https://img.webmd.com/dtmcms/live/webmd/consumer_assets/site_images/magazine/digital_issues_pdf/webmd_ma14_desktop.pdf.

Foley NM, Coll JM, Lowery AJ, Hynes SO, Kerin MJ, Sheehan M, Brodie C, Sweeney KJ. Re-appraisal of estrogen receptor negative/progesterone receptor positive (ER−/PR+) breast cancer phenotype: true subtype or technical artefact? Pathology & Oncology Research. 2018 Oct; 24: 881-4. doi: 10.1007/s12253-017-0304-5. DOI: https://doi.org/10.1007/s12253-017-0304-5

Chand P, Garg A, Singla V, Rani N. Evaluation of immunohistochemical profile of breast cancer for prognostics and therapeutic use. Nigerian Journal of Surgery. 2018 Feb; 24(2): 100-6. doi: 10.4103/njs.NJS_2_18. DOI: https://doi.org/10.4103/njs.NJS_2_18

Nielsen TO, Leung SC, Rimm DL, Dodson A, Acs B, Badve S, et al. Assessment of Ki67 in breast cancer: updated recommendations from the international Ki67 in breast cancer working group. JNCI: Journal of the National Cancer Institute. 2021 Jul; 113(7): 808-19. doi: 10.1093/jnci/djaa201. DOI: https://doi.org/10.1093/jnci/djaa201

Saleh M, Chandrashekar DS, Shahin S, Agarwal S, Kim HG, Behring M, et al. Comparative analysis of triple-negative breast cancer transcriptomics of Kenyan, African American and Caucasian Women. Translational Oncology. 2021 Jul; 14(7): 101086. doi: 10.1016/j.tranon.2021.101086. DOI: https://doi.org/10.1016/j.tranon.2021.101086

Maaruf NA, Najjar SM, Ali SA. Anatomical and Histopathological Comparative Studies Between Right and Left Mammary Tumors in a Rat Model Induced by DMBA. Pakistan Veterinary Journal. 2023 May; 43(2): 232-40. DOI: https://doi.org/10.29261/pakvetj/2023.037

Eswarachary V, Mohammed IG, Jayanna PK, Patilokaly GV, Nargund AR, Dhondalay GK, et al. HER2/neu testing in 432 consecutive breast cancer cases using FISH and IHC-A comparative study. Journal of Clinical and Diagnostic Research: JCDR. 2017 Apr; 11(4): EC01. doi: 10.7860/JCDR/2017/25625.9521. DOI: https://doi.org/10.7860/JCDR/2017/25625.9521

Hafez NH and Shaaban HM. Can GATA3 immunocytochemistry be utilized as a reliable diagnostic marker for metastatic breast carcinoma in cytological materials? A Comparative Study with mammaglobin and GCDFP-15 expression. Turk Patoloji Dergisi. 2018 May; 34(2): 143-9. doi: 10.5146/tjpath.2017.01419. DOI: https://doi.org/10.5146/tjpath.2017.01419

Park J, Oh HJ, Han D, Wang JI, Park IA, Ryu HS, et al. Parallel reaction monitoring-mass spectrometry (PRM-MS)-based targeted proteomic surrogates for intrinsic subtypes in breast cancer: comparative analysis with immunohistochemical phenotypes. Journal of Proteome Research. 2019 Nov; 19(7): 2643-53. doi: 10.1021/acs.jproteome.9b00490. DOI: https://doi.org/10.1021/acs.jproteome.9b00490

Healey MA, Hirko KA, Beck AH, Collins LC, Schnitt SJ, Eliassen AH, et al. Assessment of Ki67 expression for breast cancer subtype classification and prognosis in the Nurses’ Health Study. Breast Cancer Research and Treatment. 2017 Nov; 166: 613-22. doi: 10.1007/s10549-017-4421-3. DOI: https://doi.org/10.1007/s10549-017-4421-3

Mohanty SK, Kim SA, DeLair DF, Bose S, Laury AR, Chopra S, et al. Comparison of metastatic neuroendocrine neoplasms to the breast and primary invasive mammary carcinomas with neuroendocrine differentiation. Modern Pathology. 2016 Aug; 29(8):788-98. doi: 10.1038/modpathol.2016.69. DOI: https://doi.org/10.1038/modpathol.2016.69

Adeoye FB, Omoruyi Z, Akinbo FO. Use of Citrus limon Juice as an Alternative Antigen Retrieval Solution for Formalin Fixed Breast Cancer Tissue in Immunohistochemistry. Pan African Journal of Life Sciences. 2022 Oct; 6(3): 539-546. DOI: https://doi.org/10.36108/pajols/2202/60.0340

Triki H, Charfi S, Bouzidi L, Kridis WB, Daoud J, Chaabane K, et al. CD155 expression in human breast cancer: clinical significance and relevance to natural killer cell infiltration. Life Sciences. 2019 Aug; 231: 116543. doi: 10.1016/j.lfs.2019.116543. DOI: https://doi.org/10.1016/j.lfs.2019.116543

Hadgu E, Seifu D, Tigneh W, Bokretsion Y, Bekele A, Abebe M, et al. Breast cancer in Ethiopia: evidence for geographic difference in the distribution of molecular subtypes in Africa. BMC Women's Health. 2018 Dec; 18(1): 1-8. doi: 10.1186/s12905-018-0531-2. DOI: https://doi.org/10.1186/s12905-018-0531-2

Qadir A, Mushtaq S, Sharif MA, Mamoon N, Khadim MT. Comparison of heat induced antigen retrieval techniques for immune-histochemical estrogen receptor expression in ductal carcinoma of breast in females. Journal of College of Physicians and Surgeon Pakistan. 2010 Aug; 20: 519-23.

Grabau DA, Bendahl PO, Rydén L, Stål O, Fernö M, South and South-East Swedish Breast Cancer Groups. The prevalence of immunohistochemically determined oestrogen receptor positivity in primary breast cancer is dependent on the choice of antibody and method of heat-induced epitope retrieval–prognostic implications? Acta Oncologica. 2013 Nov; 52(8): 1657-66. doi: 10.3109/0284186X.2012.762994. DOI: https://doi.org/10.3109/0284186X.2012.762994

Abdelbadie A, Adam EA, Abdelrahman M. The effect of PT link and waterbath antigenic retrieval procedures on the expression of common receptors in breast cancer cases. Sudan Journal of Medical Sciences. 2017 Aug; 12(1): 1-8. doi: 10.18502/sjms.v12i1.853. DOI: https://doi.org/10.18502/sjms.v12i1.853