PHB2 promotes SHIP2 ubiquitination via the E3 ligase NEDD4 to regulate AKT signaling in gastric cancer.

{"sci_version": "1.2", "schema": "https://corpus.bntrg.co.uk/sci/1.0/sci.schema.json", "id": "pmid:38200519", "kind": "science-record", "revision": 3, "content_hash": "sha256:88d3c205337e00ff51d3938b1117597a942b196ff4c8f9dd32fec02018d88d91", "created_at": "2026-11-01T19:20:07Z", "updated_at": "2026-11-01T22:11:00Z", "generator": {"name": "corpus-assay-engine", "run_id": "reading-sci-2026-11-01"}, "paper": {"state": "complete", "title": "PHB2 promotes SHIP2 ubiquitination via the E3 ligase NEDD4 to regulate AKT signaling in gastric cancer.", "doi": "10.1186/s13046-023-02937-1", "pmid": "38200519", "authors": ["Liang Xu", "Wanying Xiang", "Jiezhen Yang", "Jing Gao", "Xinyue Wang", "Li Meng", "Kaihong Ye", "Xiao Hong Zhao", "Xu Dong Zhang", "Lei Jin", "Yan Ye"], "journal": "Journal of experimental & clinical cancer research : CR", "year": 2024, "abstract": "BACKGROUND: Prohibitin 2 (PHB2) exhibits opposite functions of promoting or inhibiting tumour across various cancer types. In this study, we aim to investigate its functions and underlying mechanisms in the context of gastric cancer (GC).\nMETHODS: PHB2 protein expression levels in GC and normal tissues were examined using western blot and immunohistochemistry. PHB2 expression level associations with patient outcomes were examined through Kaplan-Meier plotter analysis utilizing GEO datasets (GSE14210 and GSE29272). The biological role of PHB2 and its subsequent regulatory mechanisms were elucidated in vitro and in vivo. GC cell viability and proliferation were assessed using MTT cell viability analysis, clonogenic assays, and BrdU incorporation assays, while the growth of GC xenografted tumours was measured via IHC staining of Ki67. The interaction among PHB2 and SHIP2, as well as between SHIP2 and NEDD4, was identified through co-immunoprecipitation, GST pull-down assays, and deletion-mapping experiments. SHIP2 ubiquitination and degradation were assessed using cycloheximide treatment, plasmid transfection and co-immunoprecipitation, followed by western blot analysis.\nRESULTS: Our analysis revealed a substantial increase in PHB2 expression in GC tissues compared to adjacent normal tissues. Notably, higher PHB2 levels correlated with poorer patient outcomes, suggesting its clinical relevance. Functionally, silencing PHB2 in GC cells significantly reduced cell proliferation and retarded GC tumour growth, whereas overexpression of PHB2 further enhanced GC cell proliferation. Mechanistically, PHB2 physically interacted with Src homology 2-containing inositol 5-phosphatase 2 (SHIP2) in the cytoplasm of GC cells, thus leading to SHIP2 degradation via its novel E3 ligase NEDD4. It subsequently activated the PI3K/Akt signaling pathway and thus promoted GC cell proliferation.\nCONCLUSIONS: Our findings highlight the importance of PHB2 upregulation in driving GC progression and its association with adverse patient outcomes. Understanding the functional impact of PHB2 on GC growth contributes valuable insights into the molecular underpinnings of GC and may pave the way for the development of targeted therapies to improve patient outcomes.", "claims": [{"id": "claim-method", "statement": "PHB2 expression level associations with patient outcomes were examined through Kaplan–Meier plotter analysis utilizing GEO datasets (GSE14210 and GSE29272).", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:38200519", "locator": "sections[0] \"Abstract\" paragraph 2", "quote": "PHB2 expression level associations with patient outcomes were examined through Kaplan–Meier plotter analysis utilizing GEO datasets (GSE14210 and GSE29272)."}], "state": "complete"}, {"id": "claim-data-access", "statement": "All data supporting the findings of this study are available from the corresponding author on reasonable request.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:38200519", "locator": "sections[35] \"Availability of data and materials\" paragraph 2", "quote": "All data supporting the findings of this study are available from the corresponding author on reasonable request."}], "state": "complete"}, {"id": "claim-target-result", "statement": "PHB2 promotes gastric cancer progression by binding SHIP2, enhancing NEDD4-mediated SHIP2 degradation, and activating the PI3K/Akt pathway.", "kind": "qualitative", "target": null, "evidence": [{"asset": "source:pmid:38200519", "locator": "sections[0] \"Abstract\" paragraph 3", "quote": "Mechanistically, PHB2 physically interacted with Src homology 2-containing inositol 5-phosphatase 2 (SHIP2) in the cytoplasm of GC cells, thus leading to SHIP2 degradation via its novel E3 ligase NEDD4. It subsequently activated the PI3K/Akt signaling pathway and thus promoted GC cell proliferation."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "complete", "objective": "PHB2 promotes gastric cancer progression by binding SHIP2, enhancing NEDD4-mediated SHIP2 degradation, and activating the PI3K/Akt pathway.", "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:38200519"], "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. This lane did not download the dataset or read its licence.", "duration_hours": null, "parameters": {}, "inputs": ["data:geo:GSE14210"], "outputs": ["the source data with its repository provenance"], "instruments": [], "reagents": []}, {"id": "run_analysis", "order": 3, "name": "Run the published analysis", "kind": "analysis", "state": "complete", "blocked_on": null, "description": "Apply the paper's statistical method to the resolved series and compare the result with the published target.", "duration_hours": null, "parameters": {}, "inputs": ["data:geo:GSE14210"], "outputs": ["the replicated result and its comparison"], "instruments": [], "reagents": []}], "materials": [], "instruments": [], "reagents": [], "conditions": {}, "gaps": []}, "data": {"state": "partial", "assets": [{"id": "source:pmid:38200519", "role": "manuscript", "format": "xml", "media_type": "application/xml", "uri": "https://doi.org/10.1186/s13046-023-02937-1", "sha256": 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PHB2 expression le", "tion and co-immunoprecipitation, followed by western blot analysis. RESULTS: Our analysis revealed a s", "ive score. Two-tailed Student’s t-test. C, D Western blotting (C) and qRT-PCR (D) analysis of PHB2 express", "ollowed by Tukey’s multiple comparison. G, H Western blotting (G) and qRT-PCR (H) analysis of PHB2 in a pa"], "order_basis": "paper_text_order:383", "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": ["tissues were examined using western blot and immunohistochemistry. PHB2 expression level associations with pat", "h of GC xenografted tumours was measured via IHC staining of Ki67. The interaction among PHB2", "ted tumours was measured via IHC staining of Ki67. The interaction among PHB2 and SHIP2, as we", "rmal gastric tissues. Scale bar, 50 μm. IRS: Immunoreactive score. Two-tailed Student’s t-test. C, D Wes"], "order_basis": "paper_text_order:400", "id": "assay:2"}, {"role": "data_generation", "definition_id": "animal_model", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.77, "evidence": ["tory mechanisms were elucidated in vitro and in vivo. GC cell viability and proliferation were as"], "order_basis": "paper_text_order:677", "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": ["bility and proliferation were assessed using MTT cell viability analysis, clonogenic assays,"], "order_basis": "paper_text_order:742", "id": "assay:4"}, {"role": "data_generation", "definition_id": "transfection", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["essed using cycloheximide treatment, plasmid transfection and co-immunoprecipitation, followed by west", "e 1. List of primers. Supplementary Table 2. shRNA and siRNA sequences. Supplementary Table 3.", "of primers. Supplementary Table 2. shRNA and siRNA sequences. Supplementary Table 3. Gastric ca"], "order_basis": "paper_text_order:1170", "id": "assay:5"}, {"role": "data_generation", "definition_id": "cell_culture", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["Chinese Academy of Sciences. AGS cells were cultured in F-12 HAM’S (HyClone, USA), HGC-27, MKN-28, a", "USA), HGC-27, MKN-28, and MKN-45 cells were cultured in RPMI-1640 (HyClone, USA), and GES-1 and HEK2", "lone, USA), and GES-1 and HEK293T cells were cultured in DMEM (HyClone, USA), supplemented with 10% F"], "order_basis": "paper_text_order:2692", "id": "assay:6"}, {"role": "data_generation", "definition_id": "mass_spectrometry", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.93, "evidence": ["dates identified by immunoprecipitation plus mass spectrometry analysis in HGC-27 cells."], "order_basis": "paper_text_order:7580", "id": "assay:7"}, {"role": "analysis", "definition_id": "group_comparison", "definition_version": "f2132e96ae6c7a4e", "definition_digest": null, "required_operations": ["group_summary", "welch_t", "bootstrap_mean_difference", "group_comparison_anova"], "support_state": "supported", "source_hashes": {"dsl_catalogue": "f2132e96ae6c7a4e"}, "confidence": 0.8, "evidence": ["ANOVA"], "order_basis": "consumes_earlier_output", "id": "assay:8"}]}