Asparaginyl endopeptidase contributes to cetuximab resistance via MEK/ERK signaling in RAS wide-type metastatic colorectal cancer.

{"sci_version": "1.2", "schema": "https://corpus.bntrg.co.uk/sci/1.0/sci.schema.json", "id": "pmid:36609651", "kind": "science-record", "revision": 2, "content_hash": "sha256:9fefce7531b942f77f8fd7e56b1b0b6f8b1271e358d201057cfe949639e2f8e0", "created_at": "2026-11-01T19:01:09Z", "updated_at": "2026-11-01T21:29:50Z", "generator": {"name": "corpus-assay-engine", "run_id": "reading-sci-2026-11-01"}, "paper": {"state": "complete", "title": "Asparaginyl endopeptidase contributes to cetuximab resistance via MEK/ERK signaling in RAS wide-type metastatic colorectal cancer.", "doi": "10.1007/s12094-022-02986-6", "pmid": "36609651", "authors": ["Xiaojing Xu", "Mengling Liu", "Ke Peng", "Yiyi Yu", "Tianshu Liu"], "journal": "Clinical & translational oncology : official publication of the Federation of Spanish Oncology Societies and of the National Cancer Institute of Mexico", "year": 2023, "abstract": "BACKGROUND: Cetuximab, a monoclonal antibody targeting epidermal growth factor receptor (EGFR), is effective for RAS wild-type metastatic colorectal cancer (mCRC) patients. However, cetuximab resistance often occur and the mechanism has not been fully elucidated. The purpose of this study was to investigate the role of asparaginyl endopeptidase (AEP) in cetuximab resistance.\nMETHODS: Differentially expressed genes between cetuximab responders and non-responders were identified by analyzing the gene expression profile GSE5851, retrieved from Gene Expression Omnibus (GEO). The potential genes were further validated in cetuximab-resistant CRC cell lines. The expression of AEP in the peripheral blood and tumor tissues of mCRC patients in our hospital were detected by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry, respectively. The survival analysis was carried out by Kaplan-Meier method. The function and associated pathways of AEP were further investigated by lentivirus transfection, CCK8 assay, colony formation assay, real-time polymerase chain reaction (qPCR) and western blot.\nRESULTS: Through bioinformatics analysis, we found that the expression of AEP gene was related to progress free survival (PFS) of mCRC patients treated with cetuximab alone (P = 0.00133). The expression of AEP was significantly higher in the cetuximab-resistant CRC cell lines, as well as in mCRC patients with shorter PFS treated with cetuximab-containing therapy. Furthermore, AEP could decrease the sensitivity of CRC cells to cetuximab in vitro. And the phosphorylation level of MEK and ERK1/2 was increased in AEP overexpression cells. The downregulation of AEP using specific inhibitors could partially restore the sensitivity of CRC cells to cetuximab.\nCONCLUSION: The higher expression of AEP could contribute to the shorter PFS of cetuximab treatment in mCRC. The reason might be that AEP could promote the phosphorylation of MEK/ERK protein in the downstream signal pathway of EGFR.", "claims": [{"id": "claim-method", "statement": "The median PFS was calculated by Kaplan–Meier survival analysis, and its significance was evaluated by log rank test.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:36609651", "locator": "sections[15] \"Statistical analysis\" paragraph 2", "quote": "The median PFS was calculated by Kaplan–Meier survival analysis, and its significance was evaluated by log rank test."}], "state": "complete"}, {"id": "claim-data-access", "statement": "The gene expression profile GSE5851 was downloaded from the GEO database. GSE5851 contained 80 KRAS wide-type mCRC patients who were enrolled for treatment with cetuximab monotherapy [27]. The microarrays were conducted by Affymetrix GeneChip Scanner 3000 with GPL571 using pre-treatment tumor tissue samples.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:36609651", "locator": "sections[3] \"Microarray profile analysis from gene expression omnibus (GEO) database\" paragraph 2", "quote": "The gene expression profile GSE5851 was downloaded from the GEO database. GSE5851 contained 80 KRAS wide-type mCRC patients who were enrolled for treatment with cetuximab monotherapy [27]. The microarrays were conducted by Affymetrix GeneChip Scanner 3000 with GPL571 using pre-treatment tumor tissue samples."}], "state": "complete"}, {"id": "claim-target-result", "statement": "Analysis of the GSE5851 GEO gene-expression profile showed that higher AEP expression was associated with shorter progression-free survival in KRAS wild-type metastatic colorectal cancer patients treated with cetuximab (P = 0.00133).", "kind": "quantitative", "target": {"value": 0.00133, "unit": "p-value", "tolerance": null, "source": "Through the Gene Expression Omnibus (GEO) database, using the baseline gene expression profile (GSE5851) of KRAS wild-type mCRC patients before cetuximab treatment, we found that the level of AEP was closely related to the progression free survival (PFS) (P = 0.00133)."}, "evidence": [{"asset": "source:pmid:36609651", "locator": "sections[1] \"Introduction\" paragraph 4", "quote": "Through the Gene Expression Omnibus (GEO) database, using the baseline gene expression profile (GSE5851) of KRAS wild-type mCRC patients before cetuximab treatment, we found that the level of AEP was closely related to the progression free survival (PFS) (P = 0.00133)."}, {"asset": "source:pmid:36609651", "locator": "sections[1] \"Introduction\" paragraph 4", "quote": "Patients with higher AEP had shorter PFS."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "complete", "objective": "Analysis of the GSE5851 GEO gene-expression profile showed that higher AEP expression was associated with shorter progression-free survival in KRAS wild-type metastatic colorectal cancer patients treated with cetuximab (P = 0.00133).", "steps": [{"id": "read_published_estimate", "order": 1, "name": "Read the published estimate from the source", "kind": "other", "state": "complete", "blocked_on": null, "description": "The claim, method and evidence span published with the source article.", "duration_hours": null, "parameters": {}, "inputs": ["source:pmid:36609651"], "outputs": ["the published estimate and its evidence"], "instruments": [], "reagents": []}, {"id": "obtain_source_data", "order": 2, "name": "Obtain the source data from the named repository", "kind": "data-preparation", "state": "complete", "blocked_on": null, "description": "The GEO series accession resolves and its sample accessions are listed. 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The", "RC patients in our hospital were detected by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry, respective", "ted and detected by immunohistochemistry and ELISA, respectively. The baseline characteristics"], "order_basis": "paper_text_order:860", "id": "assay:1"}, {"role": "data_generation", "definition_id": "ihc", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["nzyme-linked immunosorbent assay (ELISA) and immunohistochemistry, respectively. The survival analysis was car", "ur department were collected and detected by immunohistochemistry and ELISA, respectively. The baseline charac", "ne characteristics are shown in Table 1. The immunohistochemical scoring was defined as follows: according to"], "order_basis": "paper_text_order:906", "id": "assay:2"}, {"role": "data_generation", "definition_id": "transfection", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.85, "evidence": ["AEP were further investigated by lentivirus transfection, CCK8 assay, colony formation assay, real-ti"], "order_basis": "paper_text_order:1089", "id": "assay:3"}, {"role": "data_generation", "definition_id": "apoptosis_assay", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.93, "evidence": ["her investigated by lentivirus transfection, CCK8 assay, colony formation assay, real-time pol"], "order_basis": "paper_text_order:1102", "id": "assay:4"}, {"role": "data_generation", "definition_id": "qpcr", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.93, "evidence": ["assay, real-time polymerase chain reaction (qPCR) and western blot. RESULTS: Through bioinfor"], "order_basis": "paper_text_order:1176", "id": "assay:5"}, {"role": "data_generation", "definition_id": "western_blot", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.93, "evidence": ["al-time polymerase chain reaction (qPCR) and western blot. RESULTS: Through bioinformatics analysis, w"], "order_basis": "paper_text_order:1185", "id": "assay:6"}, {"role": "data_generation", "definition_id": "cell_culture", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.77, "evidence": ["Sciences (Shanghai, China). Caco2 cells were cultured in Dulbecco’s Modified Eagle’s Medium (Gibco, U"], "order_basis": "paper_text_order:2800", "id": "assay:7"}, {"role": "analysis", "definition_id": "survival_analysis", "definition_version": "f2132e96ae6c7a4e", "definition_digest": null, "required_operations": ["logrank_test"], "support_state": "supported", "source_hashes": {"dsl_catalogue": "f2132e96ae6c7a4e"}, "confidence": 1.0, "evidence": ["The median PFS was calculated by Kaplan–Meier survival analysis, and its significance was evaluated by log rank test."], "order_basis": "consumes_earlier_output", "id": "assay:8"}]}