Optimal protocol for PTEN immunostaining; role of analytical and preanalytical variables in PTEN staining in normal and neoplastic endometrial, breast, and prostatic tissues
Santacana Espasa, Maria
Valls Marsal, Joan
Mirantes Barbeito, Cristina
Gatius Calderó, Sònia
García Dios, Diego Andrés
Pedersen, Hans Christian
MetadataShow full item record
In 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).
Is part ofHuman Pathology, 2014, vol. 45, núm. 3, p. 522-532
European research projects
The following license files are associated with this item:
Except where otherwise noted, this item's license is described as cc-by-nc-nd (c) Maiques et al., 2014
Showing items related by title, author, creator and subject.
An inducible knockout mouse to model the cellautonomous role of PTEN in initiating endometrial, prostate and thyroid neoplasias Mirantes Barbeito, Cristina; Eritja Sánchez, Núria; Dosil, Maria Alba; Santacana Espasa, Maria; Pallares, Judit; Gatius Calderó, Sònia; Bergadà Bertran, Laura; Maiques Carlos, Oscar; Matias-Guiu, Xavier; Dolcet Roca, Xavier (Company of Biologists, 2013)PTEN is one of the most frequently mutated tumor suppressor genes in human cancers. The role of PTEN in carcinogenesis has been validated by knockout mouse models. PTEN heterozygous mice develop neoplasms in multiple ...
Maiques Carlos, Oscar; Cuevas, Dolors; García Dios, Diego Andrés; Coenegrachts, Lieve; Santacana, Maria; Velasco, Ana; Romero, Marta; Gatius Calderó, Sònia; Lambrechts, Diether; Müller, Sven; Pedersen, Hans Christian; Dolcet Roca, Xavier; Amant, Frederic; Matias-Guiu, Xavier (Wiley, 2014)Aims: To check the usefulness of a standardized protocol of PTEN FISH in 31 endometrial carcinomas (ECs) in comparison with SNP array (SNPA), multiplex ligation-dependent probe amplification (MLPA), and immunohistochemistry. ...
Dosil Garcia, Maria Alba; Mirantes Barbeito, Cristina; Eritja Sánchez, Núria; Felip, Isidre; Navaridas Fernández de Bobadilla, Raúl; Gatius Calderó, Sònia; Santacana Espasa, Maria; Colás, Eva; Moiola, Cristian P.; Schoenenberger, Joan Antoni; Encinas Martín, Mario; Garí Marsol, Eloi; Matias-Guiu, Xavier; Dolcet Roca, Xavier (Wiley, 2017-04-28)PTEN is one of the most frequently mutated genes in human cancers. The frequency of PTEN alterations is particularly high in endometrial carcinomas. Loss of PTEN leads to dysregulation of cell division, and promotes the ...