The reliability of immunohistochemistry as a prescreening method for the diagnosis of hereditary nonpolyposis colorectal cancer (HNPCC)--results of an international collaborative study.
Hereditary nonpolyposis colorectal cancer syndrome (HNPCC) is an autosomal dominant condition accounting for 2-5% of all colorectal carcinomas as well as a small subset of endometrial, upper urinary tract and other gastrointestinal cancers. An assay to detect the underlying defect in HNPCC, inactivation of a DNA mismatch repair enzyme, would be useful in identifying HNPCC probands. Monoclonal antibodies against hMLH1 and hMSH2, two DNA mismatch repair proteins which account for most HNPCC cancers, are commercially available. This study sought to investigate the potential utility of these antibodies in determining the expression status of these proteins in paraffin-embedded formalin-fixed tissue and to identify key technical protocol components associated with successful staining. A set of 20 colorectal carcinoma cases of known hMLH1 and hMSH2 mutation and expression status underwent immunoperoxidase staining at multiple institutions, each of which used their own technical protocol. Staining for hMSH2 was successful in most laboratories while staining for hMLH1 proved problematic in multiple labs. However, a significant minority of laboratories demonstrated excellent results including high discriminatory power with both monoclonal antibodies. These laboratories appropriately identified hMLH1 or hMSH2 inactivation with high sensitivity and specificity. The key protocol point associated with successful staining was an antigen retrieval step involving heat treatment and either EDTA or citrate buffer. This study demonstrates the potential utility of immunohistochemistry in detecting HNPCC probands and identifies key technical components for successful staining.
Müller
W
W
Institute of Pathology, Heinrich Heine University, Düsseldorf, Germany.
Burgart
L J
LJ
Krause-Paulus
R
R
Thibodeau
S N
SN
Almeida
M
M
Edmonston
T B
TB
Boland
C R
CR
Sutter
C
C
Jass
J R
JR
Lindblom
A
A
Lubinski
J
J
MacDermot
K
K
Sanders
D S
DS
Morreau
H
H
Müller
A
A
Oliani
C
C
Orntoft
T
T
Ponz De Leon
M
M
Rosty
C
C
Rodriguez-Bigas
M
M
Rüschoff
J
J
Ruszkiewicz
A
A
Sabourin
J
J
Salovaara
R
R
Möslein
G
G
D048868:Adaptor Proteins, Signal Transducing; D000911:Antibodies, Monoclonal; D020137:Base Pair Mismatch; D002352:Carrier Proteins; D003123:Colorectal Neoplasms, Hereditary Nonpolyposis / Q000175:diagnosis* / Q000235:genetics / Q000276:immunology; D004260:DNA Repair; D003937:Diagnosis, Differential; D005820:Genetic Testing; D006801:Humans; D007124:Immunoenzyme Techniques / Q000592:standards* / Q000706:statistics & numerical data; D007391:International Cooperation; D007753:Laboratories / Q000592:standards; D000070957:MutL Protein Homolog 1; D009363:Neoplasm Proteins / Q000235:genetics* / Q000276:immunology; D009687:Nuclear Proteins; D015588:Observer Variation; D010375:Pedigree; D015203:Reproducibility of Results
D048868:Adaptor Proteins, Signal Transducing; D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; C501486:MLH1 protein, human; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D000070957:MutL Protein Homolog 1