Validation of specificity of antibodies for immunohistochemistry: the case of ROR2

The Wnt signalling receptor receptor tyrosine kinase-like orphan receptor 2 (ROR2) is implicated in numerous human cancers. However, there have been conflicting reports regarding ROR2 expression, some studies showing upregulation and others downregulation of ROR2 in the same cancer type. The majorit...

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Bibliographic Details
Published in:Virchows Archiv : an international journal of pathology 2017-01, Vol.470 (1), p.99-108
Main Authors: Ma, Sean S.Q., Henry, Claire E., Llamosas, Estelle, Higgins, Rupert, Daniels, Benjamin, Hesson, Luke B., Hawkins, Nicholas J., Ward, Robyn L., Ford, Caroline E.
Format: Article
Language:English
Subjects:
Antibodies - immunology
beta Catenin - genetics
Colorectal carcinoma
Colorectal Neoplasms - diagnosis
Colorectal Neoplasms - metabolism
Colorectal Neoplasms - mortality
Gene Expression Regulation, Neoplastic - genetics
Humans
Immunohistochemistry - methods
Medicine
Medicine & Public Health
Original Article
Pathology
Receptor Tyrosine Kinase-like Orphan Receptors - metabolism
Signal Transduction - physiology
Wnt Proteins - metabolism
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Summary:The Wnt signalling receptor receptor tyrosine kinase-like orphan receptor 2 (ROR2) is implicated in numerous human cancers. However, there have been conflicting reports regarding ROR2 expression, some studies showing upregulation and others downregulation of ROR2 in the same cancer type. The majority of these studies used immunohistochemistry (IHC) to detect ROR2 protein, without validation of the used antibodies. There appears to be currently no consensus on the antibody best suited for ROR2 detection or how ROR2 expression changes in various cancer types. We examined three commercially available ROR2 antibodies and found that only one bound specifically to ROR2. Another antibody cross-reacted with other proteins, and the third failed to detect ROR2 at all. ROR2 detection by IHC on 107 patient samples using the ROR2 specific antibody showed that the majority of colorectal cancers show loss of ROR2 protein. We found no association between ROR2 staining and poor patient survival, as had been previously reported. These results question the previously reported association between ROR2 and poor patient survival in colorectal cancer. Future studies should use fully validated antibodies when detecting ROR2 protein, as non-specific staining can lead to irrelevant observations and misinterpretations.
ISSN:0945-6317
1432-2307
DOI:10.1007/s00428-016-2019-5