β-Sitosterol alleviates the malignant phenotype of hepatocellular carcinoma cells via inhibiting GSK3B expression.

{"sci_version": "1.2", "schema": "https://corpus.bntrg.co.uk/sci/1.0/sci.schema.json", "id": "pmid:38814517", "kind": "science-record", "revision": 4, "content_hash": "sha256:847ae2a0b7d555b3cb6e1cac12097072897348dc1b8c52c4e731ba8f695294ec", "created_at": "2026-11-01T19:20:46Z", "updated_at": "2026-11-01T22:54:25Z", "generator": {"name": "corpus-assay-engine", "run_id": "reading-sci-2026-11-01"}, "paper": {"state": "complete", "title": "β-Sitosterol alleviates the malignant phenotype of hepatocellular carcinoma cells via inhibiting GSK3B expression.", "doi": "10.1007/s13577-024-01081-y", "pmid": "38814517", "authors": ["Ruoyu Wang", "Dan Tang", "Longyun Ou", "Jiacheng Jiang", "Yu-Nan Wu", "Xuefei Tian"], "journal": "Human cell", "year": 2024, "abstract": "To explore the effects of β-Sitosterol upon hepatocellular carcinoma cell proliferation, apoptosis, migration, invasion, and epithelial-mesenchymal transition (EMT), and to investigate the underlying mechanism using network pharmacology. Human hepatocellular carcinoma cell lines (Huh-7 and HCCLM3) were expose to gradient concentrations of β-Sitosterol (5 μg/mL, 10 μg/mL, and 20 μg/mL). Cell viability and proliferation were assessed using MTT, CCK-8, colony formation, and EdU assays.Flow cytometry was employed to evaluate cell cycle and apoptosis. Scratch and Transwell assays were performed, respectively, to detect cell migration and invasion. The levels of apoptosis-associated proteins (BAX, BCL2, and cleaved caspase3) as well as EMT-associated proteins (E-cadherin, N-cadherin, Snail, and Vimentin) were detected in Huh-7 and HCCLM3 cell lines using Western blot analysis. The drug target gene for β-Sitosterol was screened via PubChem and subsequently evaluated for expression in the GSE112790 dataset. In addition, the expression level of glycogen synthase kinase 3 beta (GSK3B) within the Cancer Genome Atlas-Liver Hepatocellular Carcinoma (TCGA-LIHC) database was analyzed, along with its correlation to the survival outcomes of patients with hepatocellular carcinoma. The diagnostic efficiency of GSK3B was assessed by analyzing the ROC curve. Subsequently, Huh-7 and HCCLM3 cell lines were transfected with the overexpression vector of GSK3B and then treated with β-Sitosterol to further validate the association between GSK3B and β-Sitosterol. GSK3B demonstrated a significantly elevated expression in patients with hepatocellular carcinoma, which could predict hepatocellular carcinoma patients' impaired prognosis based on GEO dataset and TCGA database. GSK3B inhibitor (CHIR-98014) notably inhibited cell proliferation and invasion, promoted cell apoptosis and cell cycle arrest at G0/G1 phase in hepatocellular carcinoma cells. β-Sitosterol treatment further promoted the efffects of GSK3B inhibitor on hepatocellular carcinoma cells. GSK3B overexpression has been found to enhance the proliferative and invasive capabilities of hepatocellular carcinoma cells. Furthermore it has been observed that GSK3B overexpression, it has been obsear can partially reverse the inhibitory effect of β-Sitosterol upon hepatocellular. β-Sitosterol suppressed hepatocellular carcinoma cell proliferation and invasion, and enhanced apoptosis via inhibiting GSK3B expression.", "claims": [{"id": "claim-method", "statement": "The One-way analysis of variance (ANOVA) followed Tukey’s post hoc test was performed using GraphPad Prism 8.0 software.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:38814517", "locator": "sections[13] \"Statistical analysis\" paragraph 2", "quote": "The One-way analysis of variance (ANOVA) followed Tukey’s post hoc test was performed using GraphPad Prism 8.0 software."}], "state": "complete"}, {"id": "claim-data-access", "statement": "All data supporting the findings of this study are available within the paper and its Supplementary Information.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:38814517", "locator": "sections[28] \"notes\" paragraph 1", "quote": "All data supporting the findings of this study are available within the paper and its Supplementary Information."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "complete", "objective": "", "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:38814517"], "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:GSE112790"], "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:GSE112790"], "outputs": ["the replicated result and its comparison"], "instruments": [], "reagents": []}], "materials": [], "instruments": [], "reagents": [], "conditions": {}, "gaps": []}, "data": {"state": "partial", "assets": [{"id": "source:pmid:38814517", "role": "manuscript", "format": "xml", "media_type": "application/xml", "uri": "https://doi.org/10.1007/s13577-024-01081-y", "sha256": "sha256:3447645109ad225e2dbcfe221b7bcf1938ee9b266eb66b92cccd0f0ebdcd36b3", "bytes": 36185, "access": "open", "state": "resolved", "reader": null, "reader_status": "reader_required", "identifier": "10.1007/s13577-024-01081-y", "version": null, "note": "Full article fetched from PMC; the checksum covers the rendered section body, not the original XML bytes.", "structure": {"pmcid": "PMC11194219", "source": "europe_pmc_fulltext", "coverage": "whole_body_without_references", "text_hash_scope": "rendered_section_body"}}, {"id": "data:geo:GSE112790", "role": "raw", "format": "geo-series", "media_type": "application/vnd.ncbi.geo-series+json", "uri": "https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE112790", "sha256": null, "bytes": null, "access": "open", "state": "resolved", "reader": null, "reader_status": "reader_required", "identifier": "GSE112790", "version": null, "note": "All data supporting the findings of this study are available within the paper and its Supplementary Information.", "structure": {"repository": "GEO", "access_route": "GEO accession URL", "access_statement_verbatim": "All data supporting the findings of this study are available within the paper and its Supplementary Information.", "access_statement_kind": "data_availability_statement", "sample_accessions": ["GSM3083512", "GSM3083513", "GSM3083514", "GSM3083515", "GSM3083516", "GSM3083517", "GSM3083518", "GSM3083519", "GSM3083520", "GSM3083521", "GSM3083522", "GSM3083523", "GSM3083524", "GSM3083525", "GSM3083526", "GSM3083527", "GSM3083528", "GSM3083529", "GSM3083530", "GSM3083531", "GSM3083532", "GSM3083533", "GSM3083534", "GSM3083535", "GSM3083536", "GSM3083537", "GSM3083538", "GSM3083539", "GSM3083540", "GSM3083541", "GSM3083542", "GSM3083543", "GSM3083544", "GSM3083545", "GSM3083546", "GSM3083547", "GSM3083548", "GSM3083549", "GSM3083550", "GSM3083551", "GSM3083552", "GSM3083553", "GSM3083554", "GSM3083555", "GSM3083556", "GSM3083557", "GSM3083558", "GSM3083559", "GSM3083560", "GSM3083561", "GSM3083562", "GSM3083563", "GSM3083564", "GSM3083565", "GSM3083566", "GSM3083567", "GSM3083568", "GSM3083569", "GSM3083570", "GSM3083571", "GSM3083572", "GSM3083573", "GSM3083574", "GSM3083575", "GSM3083576", "GSM3083577", "GSM3083578", "GSM3083579", "GSM3083580", "GSM3083581", "GSM3083582", "GSM3083583", "GSM3083584", "GSM3083585", "GSM3083586", "GSM3083587", "GSM3083588", "GSM3083589", "GSM3083590", "GSM3083591", "GSM3083592", "GSM3083593", "GSM3083594", "GSM3083595", "GSM3083596", "GSM3083597", "GSM3083598", "GSM3083599", "GSM3083600", "GSM3083601", "GSM3083602", "GSM3083603", "GSM3083604", "GSM3083605", "GSM3083606", "GSM3083607", "GSM3083608", "GSM3083609", "GSM3083610", "GSM3083611", "GSM3083612", "GSM3083613", "GSM3083614", "GSM3083615", "GSM3083616", "GSM3083617", "GSM3083618", "GSM3083619", "GSM3083620", "GSM3083621", "GSM3083622", "GSM3083623", "GSM3083624", "GSM3083625", "GSM3083626", "GSM3083627", "GSM3083628", "GSM3083629", "GSM3083630", "GSM3083631", "GSM3083632", "GSM3083633", "GSM3083634", "GSM3083635", "GSM3083636", "GSM3083637", "GSM3083638", "GSM3083639", "GSM3083640", "GSM3083641", "GSM3083642", "GSM3083643", "GSM3083644", "GSM3083645", "GSM3083646", "GSM3083647", "GSM3083648", "GSM3083649", "GSM3083650", "GSM3083651", "GSM3083652", "GSM3083653", "GSM3083654", "GSM3083655", "GSM3083656", "GSM3083657", "GSM3083658", "GSM3083659", "GSM3083660", "GSM3083661", "GSM3083662", "GSM3083663", "GSM3083664", "GSM3083665", "GSM3083666", "GSM3083667", "GSM3083668", "GSM3083669", "GSM3083670", "GSM3083671", "GSM3083672", "GSM3083673", "GSM3083674", "GSM3083675", "GSM3083676", "GSM3083677", "GSM3083678", "GSM3083679", "GSM3083680", "GSM3083681", "GSM3083682", "GSM3083683", "GSM3083684", "GSM3083685", "GSM3083686", "GSM3083687", "GSM3083688", "GSM3083689", "GSM3083690", "GSM3083691", "GSM3083692", "GSM3083693", "GSM3083694", "GSM3083695", "GSM3083696", "GSM3083697", "GSM3083698", "GSM3083699", "GSM3083700", "GSM3083701", "GSM3083702", "GSM3083703", "GSM3083704", "GSM3083705", "GSM3083706", "GSM3083707", "GSM3083708", "GSM3083709"], "sample_count": 198, "group_sizes": null, "group_sizes_basis": null, "platform": "570", "taxon": "Homo sapiens", "title": "Comprehensive molecular characterization of liver cancer and inheritance of the phenotypic traits during tumor recurrence [tissue]", "resolution_checked_at": "2026-11-01T15:39:54Z", "resolution_check": "identifier_resolution_only_no_download_no_licence_read", "downloaded": false, "licence_read": false}}], "gaps": ["asset:data:geo:GSE112790 not downloaded", "asset:data:geo:GSE112790 licence not read"]}, "analysis": {"state": "complete", "methods": [{"id": "method-1", "family": "group_comparison", "specific": "anova", "label": "Group comparison", "dsl_operations": ["group_summary", "welch_t", "bootstrap_mean_difference", "group_comparison_anova"], "missing_operations": [], "expressible": true, "parameters": {}, "data_shapes": ["two_or_more_groups_continuous"], "evidence": {"score": 1.0, "band": "verbatim", "quote": "The One-way analysis of variance (ANOVA) followed Tukey’s post hoc test was performed using GraphPad Prism 8.0 software."}}, {"id": "method-2", "family": "diagnostic_evaluation", "specific": "roc", "label": "Diagnostic test evaluation", "dsl_operations": ["confusion_counts", "roc_curve", "sensitivity_specificity", "solve_confusion_matrix", "classification_metrics", "published_precision_envelope"], "missing_operations": [], "expressible": true, "parameters": {}, "data_shapes": ["reference_standard_and_test"], "evidence": {"score": 0.7, "band": "medium", "quote": "ROC curve"}}], "targets": [], "gaps": []}, "capability": {"state": "complete", "requirements": [{"capability_id": "capability:computational_analysis", "label": "Computational analysis platform", "assay_families": ["meta_analysis", "machine_learning", "survival_analysis", "statistical_regression", "pca", "molecular_docking", "network_analysis", "differential_expression", "gwas"], "availability_state": "owned", "tier_unlocked": "R1", "candidate": "Automated Assays Laravel application and simulated adapters"}], "gaps": []}, "readiness": {"state": "complete", "tier": "R1", "blocking_axis": "none", "blocked_on": "nothing:replicable_now", "axes": {"method": {"tier": "R1", "expressible": true, "required_operations": ["group_summary", "welch_t", "bootstrap_mean_difference", "group_comparison_anova"], "missing_operations": [], "basis": "The paper names the method it used and the corpus's method extractor maps it onto the DSL catalogue: anova."}, "capability": {"tier": "R1", "capability_id": "capability:computational_analysis", "availability_state": "owned", "basis": "The computational analysis is owned. The physical bench the paper used is not, so a physical validation cannot be run here."}, "data": {"tier": "R1", "reachable": true, "accessions": ["GSE112790"], "resolved_accessions": ["GSE112790"], "statements": ["All data supporting the findings of this study are available within the paper and its Supplementary Information."], "basis": "The paper names GSE112790 in GEO and the accession resolves in the repository. No dataset was downloaded and no licence was read."}}, "can_run": {"dsl_operations": ["group_summary", "welch_t", "bootstrap_mean_difference", "group_comparison_anova"], "assets": ["source:pmid:38814517", "data:geo:GSE112790"], "steps": ["run_analysis"]}, "must_build": {"dsl_operations": [], "converters": [], "readers": [], "capabilities": [], "data_gaps": ["data:geo:GSE112790"]}, "gaps": []}, "review": {"state": "partial", "machine_critique": {"kind": "extraction_review", "reader": {"model": "MiniMax-M3.1-Flash", "provider": "minimax", "provider_family": "minimax", "read_at": "2026-11-01T19:20:05Z", "independence": "single_reader"}, "evidence_check": {"method": "verbatim", "availability": "verbatim"}, "calls": 5, "usage": {"prompt_tokens": 10576, "completion_tokens": 1797, "cached_tokens": 256}}, "human_critique": null, "stages": [{"id": "extraction", "state": "complete", "reader": "MiniMax-M3.1-Flash", "provider": "minimax"}], "gaps": ["review.independent_reader", "review.machine_critique", "review.human_critique"]}, "execution": {"state": "absent", "status": "not_run", "mode": null, "runs": [], "gaps": ["execution.not_run"]}, "provenance": {"entities": [{"id": "pmid:38814517", "type": "paper", "label": "pmid:38814517"}, {"id": "source:pmid:38814517", "type": "manuscript", "label": "PMID 38814517 full text"}, {"id": "data:geo:GSE112790", "type": "dataset", "label": "GEO GSE112790"}], "activities": [{"id": "activity-reading-sci-2026-11-01", "type": "classification", "used": [{"id": "pmid:38814517", "type": "paper", "label": "pmid:38814517"}, {"id": "source:pmid:38814517", "type": "manuscript", "label": "PMID 38814517 full text"}, {"id": "data:geo:GSE112790", "type": "dataset", "label": "GEO GSE112790"}], "agent": "agent:corpus-assay-engine"}, {"id": "activity-read-38814517", "type": "extraction", "used": ["source:pmid:38814517"], "agent": "agent:minimax-reader"}, {"id": "activity-resolve-GSE112790", "type": "accession_resolution", "used": ["data:geo:GSE112790"], "agent": "agent:corpus-assay-engine"}], "agents": [{"id": "agent:corpus-assay-engine", "type": "software", "label": "Corpus assay engine"}, {"id": "agent:minimax-reader", "type": "software", "label": "MiniMax literature reader"}], "source_run_id": "reading-sci-2026-11-01", "dsl_catalogue_fingerprint": "343fb30756ba6ec3", "capability_table_fingerprint": "2cb3e42f047d2ecf", "snapshot_id": "ba34b49490d9bfaf"}, "lineage": {"parent_id": "pmid:38814517", "parent_hash": "sha256:20c5b5eda6d4b408db7b038e7c28b7d84ea7d3a3f3b5501190b476f0450bf805", "fork_of": null, "generation": 3}, "gaps": ["asset:data:geo:GSE112790 licence not read", "asset:data:geo:GSE112790 not downloaded", "execution.not_run", "review.human_critique", "review.independent_reader", "review.machine_critique"], "assays": [{"role": "data_generation", "definition_id": "apoptosis_assay", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["hepatocellular carcinoma cell proliferation, apoptosis, migration, invasion, and epithelial-mesench", "etry was employed to evaluate cell cycle and apoptosis. Scratch and Transwell assays were performed", "t cell migration and invasion. The levels of apoptosis-associated proteins (BAX, BCL2, and cleaved", "ll proliferation and invasion, promoted cell apoptosis and cell cycle arrest at G0/G1 phase in hepa"], "order_basis": "paper_text_order:159", "id": "assay:1"}, {"role": "data_generation", "definition_id": "flow_cytometry", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.93, "evidence": ["MTT, CCK-8, colony formation, and EdU assays.Flow cytometry was employed to evaluate cell cycle and apop"], "order_basis": "paper_text_order:557", "id": "assay:2"}, {"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": ["etected in Huh-7 and HCCLM3 cell lines using Western blot analysis. The drug target gene for β-Sitoste"], "order_basis": "paper_text_order:931", "id": "assay:3"}, {"role": "data_generation", "definition_id": "clinical_cohort", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["its correlation to the survival outcomes of patients with hepatocellular carcinoma. The diagnostic eff", "rated a significantly elevated expression in patients with hepatocellular carcinoma, which could predic"], "order_basis": "paper_text_order:1315", "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": 0.85, "evidence": ["bsequently, Huh-7 and HCCLM3 cell lines were transfected with the overexpression vector of GSK3B and"], "order_basis": "paper_text_order:1477", "id": "assay:5"}, {"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": 1.0, "evidence": ["The One-way analysis of variance (ANOVA) followed Tukey’s post hoc test was performed using GraphPad Prism 8.0 software."], "order_basis": "consumes_earlier_output", "id": "assay:6"}]}