PWE-163 Chemr23 and BLT1 Receptor Expression in Colorectal Cancer
Bibliographic record
Abstract
Introduction Eicosapentaenoic acid (EPA) is an omega-3 polyunsaturated fatty acid which has anti-colorectal cancer (CRC) activity. The molecular mechanism (s) underlying the anti-neoplastic activity of EPA are not understood. Trihydroxy-EPA, also known as Resolvin E1 (RvE1), is an oxygenated derivative of EPA, that has been shown to inhibit NK-κB signalling, which is implicated in colorectal carcinogenesis. RvE1 has been shown to bind to two G-protein coupled receptors, ChemR23 and BLT1. We investigated whether ChemR23 and BLT1 receptors are expressed in human CRC. Methods Seven human CRC cell lines (HCA7, LoVo, T84, HRT18, HT29, Caco2 and HCT116) were characterised for ChemR23 and BLT1 expression by quantitative real-time polymerase chain reaction, western blotting and immunofluorescence. Jurkat and THP-1 cells were used as positive controls for ChemR23 and BLT1, respectively. Membrane protein fraction analysis was carried out using a transmembrane protein extraction kit. Densitometric analysis was performed using BIO-RAD Quantity One Software. Human CRC tissue was examined for ChemR23 expression by immunohistochemistry on formalin-fixed, paraffin-embedded tissue blocks. Results ChemR23 and BLT1 messenger RNA expression was detected in all seven human CRC cell lines. ChemR23 protein (45kDa) expression was also observed in all human CRC cell lines, with Caco2 cells expressing around two-fold more ChemR23 receptor protein relative to a-tubulin than other CRC cell lines. However, BLT1 receptor protein was not detected in any of the human CRC cell lines, but was confirmed in monocytic THP-1 cells (38kDa). ChemR23 protein was enriched in the membrane protein fraction of Caco2 cells. ChemR23 protein levels increased with confluency in Caco2 cells. There was a three-fold increase in ChemR23 protein expression in 100% confluent Caco2 cells compared with less confluent cell cultures. In contrast, HCA-7 cells did not display confluence-dependent changes in ChemR23 protein expression. Immunofluorescence demonstrated predominant cytoplasmic localisation of ChemR23 with a heterogeneous population of ChemR23-expressing and negative cells. ChemR23 immunohistochemistry on primary CRC tissue demonstrated homogeneous ChemR23 immunoreactivity in CRC cells with some stromal cell staining, including endothelial cells. Conclusion ChemR23 (but not BLT1) protein is expressed by human CRC cells (particularly Caco2) in vitro and in cancer cells in human primary CRCs. ChemR23 protein expression varies in vitro in a confluence-dependent manner, with heterogeneous expression by Caco2 cells. ChemR23 is localised predominantly in cancer cells in human CRC. Investigation of ChemR23-dependent anti-CRC activity of RvE1 is warranted. Disclosure of Interest None Declared.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.001 | 0.001 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.006 | 0.001 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".