TY - JOUR
T1 - Correlation of genetic and immunodetection of TP53 mutations in malignant and benign prostate tissues
AU - Wertz, Ingrid E.
AU - Deitch, Arline D.
AU - Gumerlock, Paul H.
AU - Gandour-Edwards, Regina
AU - Chi, Sung Gil
AU - De Vere White, Ralph W.
N1 - Funding Information:
From the Department of Urology, the Cancer and Molecular Research Laboratory, Division of Hematology/Oncology, Department of Internal Medicine, and the Department of Pathology, University of California, Davis, Sacramento, CA. Accepted for publication December 4, 1995. Supported in part by the RobertJ. Mathews Foundation for Pros-rate Cancer Research, and by grant nos. PO1-CA55792, UO1-CA57183, and UO1-CA60098 from the National Cancer Institute. Address correspondence and reprint requests to Ralph W. de Vere White, 4301 X St, Suite 2220, Sacramento, CA 95817. Copyright © 1996 by W.B. Saunders Company 0046-8177/96/2706-001555.00/0 tion of microwave heat-induced antigen unmasking and a cocktail of the DO-1 and DO-7 antibodies. This approach identified 14 of 15 (93%) cell lines and patient samples having TP53 missense mutations in the exons 5 to 8 region. Of the 21 patient samples and cell lines that were either normal by SSCP or expressed p53 mutations that are not expected to stain, 18 (86%) were immunonegative. Because of this good correlation between molecular and IHC analysis, this approach may help to resolve the uncertainty about TP53 in CaP tumorigenesis. HUM PATHOL 27:573--580. Copyright © 1996 byW.B. Saunders Company Key words: prostatic neoplasms, TP53, p53, immtmohistochemis-try, antigen unmasking, single strand conformational polymorphism. Abbreviations: CaP, prostatic adenocarcinoma; TP53, p53 tumor suppressor gene, IHC, immunohistoehemical; BPH, benign prostatic hyperplasia; SSCP, single-strand eonformational polymorphism; PCR, polymerase chain reaction; H~O2, hydrogen peroxide; PBS, phosphate-buffered saline.
PY - 1996
Y1 - 1996
N2 - The prognostic value of the p53 gene (TP53), the most commonly mutated gene in human cancers, has been well established for several cancer types. However, because varying frequencies of TP53 mutations have been identified in prostatic adenocarcinoma (CaP) by genetic and immunohistochemical (IHC) studies, the role of TP53 in CaP tumorigenesis is currently unresolved. These experimental discrepancies could be caused by tissue heterogeneity within prostatic neoplasms, variations in experimental protocols, or other factors. Thus, the goal of this study was to develop a reliable IHC approach for the detection of p53 in archival prostate tissue. The authors evaluated four p53 antibodies, CM-1, 1801, DO-1, and DO-7, for their ability to reveal p53. They chose two reference CaP cell lines, 26 patient specimens (including eight benign prostatic hyperplasias (BPHs), 16 CaPs, and two lymph node metastases), one prostate and nine kidney cell lines for p53 analysis. The TP53 status of these samples was characterized using single-strand conformational polymorphism (SSCP) analysis of RNA/PCR products and sequencing. IHC detection of p53 was markedly enhanced by using the combination of microwave heat-induced antigen unto g anti a cocktail of the DO-1 and DO-7 antibodies. This approach identified 14 of 15 (93%) cell lines and patient samples having TP53 missense mutations in the exons 5 to 8 region. Of the 21 patient samples and cell lines that were either normal by SSCP or expressed p53 mutations that are not expected to stain, 18 (86%) were immunonegative. Because of this good correlation between molecular and IHC analysis, this approach may help to resolve the uncertainty about TP53 in CaP tumorigenesis.
AB - The prognostic value of the p53 gene (TP53), the most commonly mutated gene in human cancers, has been well established for several cancer types. However, because varying frequencies of TP53 mutations have been identified in prostatic adenocarcinoma (CaP) by genetic and immunohistochemical (IHC) studies, the role of TP53 in CaP tumorigenesis is currently unresolved. These experimental discrepancies could be caused by tissue heterogeneity within prostatic neoplasms, variations in experimental protocols, or other factors. Thus, the goal of this study was to develop a reliable IHC approach for the detection of p53 in archival prostate tissue. The authors evaluated four p53 antibodies, CM-1, 1801, DO-1, and DO-7, for their ability to reveal p53. They chose two reference CaP cell lines, 26 patient specimens (including eight benign prostatic hyperplasias (BPHs), 16 CaPs, and two lymph node metastases), one prostate and nine kidney cell lines for p53 analysis. The TP53 status of these samples was characterized using single-strand conformational polymorphism (SSCP) analysis of RNA/PCR products and sequencing. IHC detection of p53 was markedly enhanced by using the combination of microwave heat-induced antigen unto g anti a cocktail of the DO-1 and DO-7 antibodies. This approach identified 14 of 15 (93%) cell lines and patient samples having TP53 missense mutations in the exons 5 to 8 region. Of the 21 patient samples and cell lines that were either normal by SSCP or expressed p53 mutations that are not expected to stain, 18 (86%) were immunonegative. Because of this good correlation between molecular and IHC analysis, this approach may help to resolve the uncertainty about TP53 in CaP tumorigenesis.
KW - TP53
KW - antigen unmasking
KW - immunohistochemistry
KW - p53
KW - prostatic neoplasms
KW - single strand conformational polymorphism
UR - http://www.scopus.com/inward/record.url?scp=0029948503&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0029948503&partnerID=8YFLogxK
U2 - 10.1016/S0046-8177(96)90164-1
DO - 10.1016/S0046-8177(96)90164-1
M3 - Article
C2 - 8666367
AN - SCOPUS:0029948503
SN - 0046-8177
VL - 27
SP - 573
EP - 580
JO - Human Pathology
JF - Human Pathology
IS - 6
ER -
