The effect of PT link and waterbath antigenic retrieval procedures on the expression of common receptors in breast cancer cases

Ahmed  Abdelbadie; Elsadig  Ahmed Adam; Manar  Abdelrahman

doi:10.52981/sjms.v12i1.1218

Ahmed Abdelbadie
Elsadig Ahmed Adam
Manar Abdelrahman

DOI: https://doi.org/10.52981/sjms.v12i1.1218

Keywords: Antigen retrieval, ER, PR, HER2 expression, PT link, waterbath

Abstract

Background: Antigen retrieval (AR) techniques play a major role in determining the quality and functional state of tissue proteins recovered from formalin fixed paraffin embedded blocks (FFPEB). Different techniques are used each with its merits and drawbacks.

Materials and Method: FFPEB from cases diagnosed as breast cancer were examined for the expression of estrogen- receptor (ER), progesterone receptor (PR) and HER2 by immunohistochemical techniques using two different AR methods.

Results: Fifty cases, 49 were from females, the age ranged from 28–85 years were examined. The expression of ER using water bath and PT link (pretreatment system) as antigen retrieval methods was equal, both methods showed 20(40%) positive cases and 30 (60%) negative cases, whereas, the expression of PR was found positive in 18 cases (36%) and negative in 32 (64%) when water bath was used, in comparison to 10 positive (20%) and 40 negative (80%) when PT link was used (P value 0.312). The expression of HER2 was as follows; water bath HER2 weak positive 20 cases (40%), moderate expression 10 cases (20%), overexpression 2 cases (4%). PT link weak expression 13 cases (26%), moderate expression 15 cases (30%), overexpression 4cases (8%); yet, the total number of negative cases and positive cases was equal for both methods (p-value 0.035).

Conclusion: Antigen retrieval methods were found to affect the expression of common receptors in breast cancer. The magnitude of this effect was found to be significant in PR and HER2 receptors, however, the different methods of antigen retrieval did not affect the expression of ER. The appropriate and suitable retrieval methods have to be chosen individually for each receptor.

References

[1] H. Yamashita, Y. Yando, M. Nishio et al., “Immunohistochemical evaluation ofhormone receptor status for predicting response to endocrine therapy in metastaticbreast cancer,” Breast cancer (Tokyo, Japan), vol. 13, no. 1, pp. 74–83, 2006.
[2] H. Yamashita, “Current research topics in endocrine therapy for breast cancer,”International Journal of Clinical Oncology, vol. 13, no. 5, pp. 380–383, 2008.
[3] L. A. Carey, C. M. Perou, C. A. Livasy et al., “Race, breast cancer subtypes, andsurvival in the Carolina Breast Cancer Study,” The Journal of the American MedicalAssociation, vol. 295, no. 21, pp. 2492–2502, 2006.
[4] H. Y. Chang, D. S. A. Nuyten, J. B. Sneddon et al., “Robustness, scalability, andintegration of a wound-response gene expression signature in predicting breastcancer survival,” Proceedings of the National Academy of Sciences of the UnitedStates of America, vol. 102, no. 10, pp. 3738–3743, 2005.
[5] M. Dolled-Filhart, L. Rydén, M. Cregger et al., “Classification of breast cancer usinggenetic algorithms and tissue microarrays,” Clinical Cancer Research, vol. 12, no. 21,pp. 6459–6468, 2006.
[6] T. Sørlie, R. Tibshirani, and J. Parker, “Repeated observation of breast tumorsubtypes in independent gene expression data sets,” Proceedings of the NationalAcademy of Sciences of the United States of America, vol. 100, pp. 8418–8423, 2003.
[7] K. Dahlman-Wright, V. Cavailles, S. A. Fuqua et al., “International union ofpharmacology. LXIV. Estrogen receptors,” Pharmacological Reviews, vol. 58, no. 4, pp. 773–781, 2006.
[8] E. R. Levin, “Integration of the extranuclear and nuclear actions of estrogen,”Molecular Endocrinology, vol. 19, no. 8, pp. 1951–1959, 2005.
[9] B. J. Deroo and K. S. Korach, “Estrogen receptors and human disease,” Journal of Clinical Investigation, vol. 116, no. 3, pp. 561–570, 2006.
[10] M. Misrahi, M. Atger, L. d’Auriol et al., “Complete amino acid sequence of the humanprogesterone receptor deduced from cloned cDNA,” Biochemical and BiophysicalResearch Communications, vol. 143, no. 2, pp. 740–748, 1987.
[11] G. Kase Nathan, Leon;. Glass, and L. Robert, “Clinical gynecologic endocrinology andinfertility,” in Hagerstown, MD: Lippincott, Williams & Wilkins, Hagerstown, 1999,ISBN 0-683-30379-1.
[12] M. Tan and D. Yu, “Molecular mechanisms of ErbB2-mediated breast cancerchemoresistance,” Advances in Experimental Medicine and Biology, vol. 608, pp.119–129, 2007.
[13] K. Emoto, S. Yamashita, and Y. Okada, “Mechanisms of heat-induced antigenretrieval: Does pH or ionic strength of the solution play a role for refoldingantigens?” Journal of Histochemistry and Cytochemistry, vol. 53, no. 11, pp. 1311–1321, 2005.
[14] E. M. Elgaili, D. O. Abuidris, M. Rahman, A. M. Michalek, and S. I. Mohammed, “Breastcancer burden in central Sudan,” International Journal of Women’s Health, vol. 2, no.1, pp. 77–82, 2010.