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BerEP4 and AE1/3 are Reliable Markers of Epithelial Content for Biomarker Discovery Using Reverse Phase Protein Arrays (RPPA)
| Content Provider | Semantic Scholar |
|---|---|
| Author | Negm, Ola H. |
| Copyright Year | 2016 |
| Abstract | Background: Reverse phase protein array (RPPA) allows precise protein quantification. Use of homogenised samples confounds interpretation due to uncertainty of cellular content. We hypothesised that antibodies AE1/3 (Type I/II keratins) and BerEP4 (EpCAM) could help to quantify epithelium in tissue from colorectal cancers (CRC). Methods: AE1/3 and BerEP4 expression was quantified using RPPA and immunohistochemistry (IHC) in formalin-fixed paraffinembedded tissues from 19 CRCs. For RPPA, samples were tested in sextuplicate. IHC was quantified by transforming images into positive/negative pixels using ImageJ. Results: In RPPA, the BerEP4 expression was greater than AE1/3 (p < 0.001). There was excellent precision with a mean coefficient of variation of 5.9% for AE1/3 and 6.9% for BerEP4. Expression of AE1/3 and BerEP4 showed significant correlation (p < 0.0001). For IHC, antibodies showed specific epithelial staining. There was no difference in intensity of expression but there were slight differences in extent of expression with, on average, 3.5% more AE1/3-positive pixels than BerEP4-positive pixels (p = 0.016). Antibody expression (number of positive pixels) showed significant correlation (p < 0.0001). Comparison of RPPA with IHC showed significant correlation for each antibody, (BerEP4: p = 0.044; AE1/3: p = 0.037). Combining markers (using the geometric mean) improved the correlation between the assays (p= 0.028). Conclusion: Expression of AE1/3 and BerEP4 correlate with each other in both RPPA and IHC. For each antibody, the two assays show significant correlation. Either antibody can be used to quantify tumour epithelium in RPPA with improved performance if markers are combined. Normalising RPPA data to “epithelial housekeepers” may improve biomarker data interpretation. |
| File Format | PDF HTM / HTML |
| DOI | 10.23937/2469-5807/1510031 |
| Volume Number | 2 |
| Alternate Webpage(s) | https://clinmedjournals.org/articles/ijpcr/international-journal-of-pathology-and-clinical-research-ijpcr-2-031.pdf |
| Alternate Webpage(s) | https://doi.org/10.23937/2469-5807%2F1510031 |
| Language | English |
| Access Restriction | Open |
| Content Type | Text |
| Resource Type | Article |
