Improved immunohistochemical detection of type 1 insulin-Like growth factor receptor in human tumors
<p><strong>Background:</strong> Insulin-like growth factors (IGFs) are known to play important roles in cancer biology, prompting evaluation of drugs targeting type 1 IGF receptor (IGF-1R). However, there is considerable lack of consensus in immunohistochemical (IHC) studies of IGF...
| Main Authors: | , , , , , |
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| Format: | Journal article |
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OMICS International Journals
2016
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| _version_ | 1797082452500414464 |
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| author | Macaulay, V Aleksic, T Turley, H Worrall, A Verrill, C Campo, L |
| author_facet | Macaulay, V Aleksic, T Turley, H Worrall, A Verrill, C Campo, L |
| author_sort | Macaulay, V |
| collection | OXFORD |
| description | <p><strong>Background:</strong> Insulin-like growth factors (IGFs) are known to play important roles in cancer biology, prompting evaluation of drugs targeting type 1 IGF receptor (IGF-1R). However, there is considerable lack of consensus in immunohistochemical (IHC) studies of IGF-1R in human tumors, confounding attempts to assess the predictive and prognostic significance of IGF-1R expression and subcellular localization. Likely sources of variation include use of different IGF-1R polyclonal antibodies and methods for IHC. Here, we aimed to develop a robust IGF-1R IHC protocol using a monoclonal antibody, suitable for use in formalin-fixed paraffin-embedded (FFPE) tissues.</p> <p><strong>Methods:</strong> Using controls including samples of FFPE tissues and tumor cells of defined IGF-1R expression, we used IHC and western blotting to compare polyclonal antibody #3027 with monoclonals #9750 and #14534 (Cell Signaling Technology).</p> <p><strong>Results:</strong> Compared with #3027, the monoclonals exhibited superior discrimination between IGF-1R-high and IGF-1R-deficient cells in manual IHC, signal generated by #9750 reflecting differences in IGF-1R expression detected by western blotting. In tissues, IGF-1R detected by #14534 was predominantly plasma membrane-associated, while #9750 detected IGF-1R in the plasma membranes, cytoplasm and nucleus of prostate and renal cancers, recapitulating appearances we described using previous lots of #3027, and reflecting subcellular localizations reported using other techniques. Use of #9750 and #14534 in an autostainer showed adequate differentiation of high vs low IGF-1R cells, but did not recapitulate appearances of manually-stained tissues. We provide a detailed protocol for the preferred manual method using #9750.</p> <p><strong>Conclusion:</strong> Standardization of IGF-1R IHC will promote understanding of the role of IGF-1R in tumor biology, and its potential as a candidate prognostic and predictive biomarker.</p> |
| first_indexed | 2024-03-07T01:28:18Z |
| format | Journal article |
| id | oxford-uuid:92b7d9e8-c87e-4834-be0d-e9b63bbb0b4c |
| institution | University of Oxford |
| last_indexed | 2024-03-07T01:28:18Z |
| publishDate | 2016 |
| publisher | OMICS International Journals |
| record_format | dspace |
| spelling | oxford-uuid:92b7d9e8-c87e-4834-be0d-e9b63bbb0b4c2022-03-26T23:27:31ZImproved immunohistochemical detection of type 1 insulin-Like growth factor receptor in human tumorsJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:92b7d9e8-c87e-4834-be0d-e9b63bbb0b4cSymplectic Elements at OxfordOMICS International Journals2016Macaulay, VAleksic, TTurley, HWorrall, AVerrill, CCampo, L<p><strong>Background:</strong> Insulin-like growth factors (IGFs) are known to play important roles in cancer biology, prompting evaluation of drugs targeting type 1 IGF receptor (IGF-1R). However, there is considerable lack of consensus in immunohistochemical (IHC) studies of IGF-1R in human tumors, confounding attempts to assess the predictive and prognostic significance of IGF-1R expression and subcellular localization. Likely sources of variation include use of different IGF-1R polyclonal antibodies and methods for IHC. Here, we aimed to develop a robust IGF-1R IHC protocol using a monoclonal antibody, suitable for use in formalin-fixed paraffin-embedded (FFPE) tissues.</p> <p><strong>Methods:</strong> Using controls including samples of FFPE tissues and tumor cells of defined IGF-1R expression, we used IHC and western blotting to compare polyclonal antibody #3027 with monoclonals #9750 and #14534 (Cell Signaling Technology).</p> <p><strong>Results:</strong> Compared with #3027, the monoclonals exhibited superior discrimination between IGF-1R-high and IGF-1R-deficient cells in manual IHC, signal generated by #9750 reflecting differences in IGF-1R expression detected by western blotting. In tissues, IGF-1R detected by #14534 was predominantly plasma membrane-associated, while #9750 detected IGF-1R in the plasma membranes, cytoplasm and nucleus of prostate and renal cancers, recapitulating appearances we described using previous lots of #3027, and reflecting subcellular localizations reported using other techniques. Use of #9750 and #14534 in an autostainer showed adequate differentiation of high vs low IGF-1R cells, but did not recapitulate appearances of manually-stained tissues. We provide a detailed protocol for the preferred manual method using #9750.</p> <p><strong>Conclusion:</strong> Standardization of IGF-1R IHC will promote understanding of the role of IGF-1R in tumor biology, and its potential as a candidate prognostic and predictive biomarker.</p> |
| spellingShingle | Macaulay, V Aleksic, T Turley, H Worrall, A Verrill, C Campo, L Improved immunohistochemical detection of type 1 insulin-Like growth factor receptor in human tumors |
| title | Improved immunohistochemical detection of type 1 insulin-Like growth factor receptor in human tumors |
| title_full | Improved immunohistochemical detection of type 1 insulin-Like growth factor receptor in human tumors |
| title_fullStr | Improved immunohistochemical detection of type 1 insulin-Like growth factor receptor in human tumors |
| title_full_unstemmed | Improved immunohistochemical detection of type 1 insulin-Like growth factor receptor in human tumors |
| title_short | Improved immunohistochemical detection of type 1 insulin-Like growth factor receptor in human tumors |
| title_sort | improved immunohistochemical detection of type 1 insulin like growth factor receptor in human tumors |
| work_keys_str_mv | AT macaulayv improvedimmunohistochemicaldetectionoftype1insulinlikegrowthfactorreceptorinhumantumors AT aleksict improvedimmunohistochemicaldetectionoftype1insulinlikegrowthfactorreceptorinhumantumors AT turleyh improvedimmunohistochemicaldetectionoftype1insulinlikegrowthfactorreceptorinhumantumors AT worralla improvedimmunohistochemicaldetectionoftype1insulinlikegrowthfactorreceptorinhumantumors AT verrillc improvedimmunohistochemicaldetectionoftype1insulinlikegrowthfactorreceptorinhumantumors AT campol improvedimmunohistochemicaldetectionoftype1insulinlikegrowthfactorreceptorinhumantumors |