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dc.contributor.authorMaiques Carlos, Oscar
dc.contributor.authorSantacana Espasa, Maria
dc.contributor.authorValls Marsal, Joan
dc.contributor.authorPallares, Judit
dc.contributor.authorMirantes Barbeito, Cristina
dc.contributor.authorGatius Calderó, Sònia
dc.contributor.authorGarcía Dios, Diego Andrés
dc.contributor.authorAmant, Frederic
dc.contributor.authorPedersen, Hans Christian
dc.contributor.authorDolcet Roca, Xavier
dc.contributor.authorMatias-Guiu, Xavier
dc.description.abstractIn some tumors, phosphatase and tensin homolog (PTEN) inactivation may have prognostic importance and predictive value for targeted therapies. Immunohistochemistry (IHC) may be an effective method to demonstrate PTEN loss. It was claimed that PTEN IHC showed poor reproducibility, lack of standardization, and variable effects of preanalytical factors. In this study, we developed an optimal protocol for PTEN IHC, with clone 6H2.1, by checking the relevance of analytical variables in normal tissue and tumors of endometrium, breast, and prostate. Pattern and intensity of cellular staining and background nonspecific staining were quantified and subjected to statistical analysis by linear mixed models. The proposed protocol showed a statistically best performance (P < .05) and included a high target retrieval solution, 1:100 primary antibody dilution (2.925 mg/L), FLEX diluent, and EnVisionFLEX+ detection method, with a sensitivity and specificity of 72.33% and 78.57%, respectively. Staining specificity was confirmed in cell lines and animal models. Endometrial carcinomas with PTEN genetic abnormalities showed statistically lower staining than tumors without alterations (mean histoscores, 34.66 and 119.28, respectively; P = .01). Controlled preanalytical factors (delayed fixation and overfixation) did not show any statistically significant effect on staining with optimal protocol (P > .001). However, there was a trend of significance for decreased staining and fixation under high temperature. Moreover, staining was better in endometrial aspirates than in matched hysterectomy specimens, subjected to less controlled preanalytical variables (mean histoscores, 80 and 40, respectively; P = .002). A scoring system combining intensity of staining and percentage of positive cells was statistically associated with PTEN alterations (P = .01).ca_ES
dc.description.sponsorshipThe study was done according to the research collaboration with Dako Denmark A/S. The research team was also supported by grants FIS PI100922, Fundación Mutua Madrileña AP75732010, 2009SGR794, RD12/0036/0013, Fundación Asociación Española contra el Cancer, programa de intensificación de la investigación, Instituto Carlos III, Verelst Baarmoederkankerfonds, Leuven, and European Network for Individualized Treatment of Endometrial Carcinoma. F. A. is senior researcher for the research fund Flandersb. Tumor samples were obtained with the support of XarxaCatalana de Bancs de Tumors, the TumorBanc Platform of RTICC, and Red de Biobancos (RD09/0076/00059)ca_ES
dc.relation.isformatofReproducció del document publicat a
dc.relation.ispartofHuman Pathology, 2014, vol. 45, núm. 3, p. 522-532ca_ES
dc.rightscc-by-nc-nd (c) Maiques et al., 2014ca_ES
dc.subjectPreanalytical variablesca_ES
dc.titleOptimal protocol for PTEN immunostaining; role of analytical and preanalytical variables in PTEN staining in normal and neoplastic endometrial, breast, and prostatic tissuesca_ES

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