Best practice for PTEN gene and protein assessment in anatomic pathology

Best practice for PTEN gene and protein assessment in anatomic pathology

Author Carvalho, Katia C. Google Scholar
Maia, Beatriz M. Google Scholar
Omae, Samantha V. Google Scholar
Rocha, Antonio A. Autor UNIFESP Google Scholar
Covizzi, Luiz P. Google Scholar
Vassallo, Jose Google Scholar
Rocha, Rafael M. Google Scholar
Soares, Fernando A. Google Scholar
Institution Universidade de São Paulo (USP)
Hosp AC Camargo Fund Antonio Prudente
Universidade Federal de São Paulo (UNIFESP)
Abstract There is a lack of standardization of a best practice protocol for Phosphatase and Tensin Homolog (PTEN) assessment by immunohistochemistry in anatomic pathology routine practice. We performed immunohistochemistry for 19 antibodies against PTEN, eleven of which were excluded during the standardization step. Immunohistochemistry of the remaining eight antibodies was performed on a Tissue Microarray containing 55 prostate and 40 renal carcinoma samples. Fluorescent in situ hybridization (FISH) was used as reference standard for immunohistochemistry specificity evaluation. Concerning nuclear staining, polyclonal (Cat#22034-1-AP); 6H2.1 mMAb (Cat#ABM-2052), Y184 RabMAb (Cat#NB110-57441) and 217702 mMAb antibodies presented the highest agreement with fluorescent in situ hybridization (p<0.001 for all) and with regard to cytoplasmic staining, Y184 RabMAb (Cat#NB110-57441); polyclonal (Cat#22034-1-AP) and 217702 mMAb presented the highest agreement (p < 0.001 for all). Our results indicate that several commercially available antibodies do not show reliability of sensitivity and specificity for PTEN evaluation and we propose 6H2.1 mMAb (Cat#ABM-2052) as the antibody of choice for laboratory standardization and best practice in clinical routine, which demonstrated excellent sensitivity for both nuclear and cytoplasmic staining, specificity for PTEN by Western blot and good correlation with PTEN status by FISH with regard to nuclear staining. (C) 2013 Elsevier GmbH. All rights reserved.
Keywords PTEN
Immunohistochemistry
Fluorescent in situ hybridization
Human
Prostatic neoplasm
Renal cell carcinoma
Language English
Sponsor Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Date 2014-01-01
Published in Acta Histochemica. Jena: Elsevier Gmbh, Urban & Fischer Verlag, v. 116, n. 1, p. 25-31, 2014.
ISSN 0065-1281 (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 25-31
Origin http://dx.doi.org/10.1016/j.acthis.2013.04.013
Access rights Closed access
Type Article
Web of Science ID WOS:000331671300005
URI http://repositorio.unifesp.br/handle/11600/37234

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