MiR-16-5p suppresses breast cancer proliferation by targeting ANLN.

{"sci_version": "1.2", "schema": "https://corpus.bntrg.co.uk/sci/1.0/sci.schema.json", "id": "pmid:34743685", "kind": "science-record", "revision": 5, "content_hash": "sha256:bf2c6fd2823431ddd5a5a7afd69fba46d79f37ae0b94607956234b50f88fb58e", "created_at": "2026-11-01T17:36:25Z", "updated_at": "2026-11-01T22:47:49Z", "generator": {"name": "corpus-assay-engine", "run_id": "reading-sci-2026-11-01"}, "paper": {"state": "complete", "title": "MiR-16-5p suppresses breast cancer proliferation by targeting ANLN.", "doi": "10.1186/s12885-021-08914-1", "pmid": "34743685", "authors": ["Ziming Wang", "Siyuan Hu", "Xinyang Li", "Zhiwei Liu", "Danyang Han", "Yukun Wang", "Limin Wei", "Guangping Zhang", "Xinshuai Wang"], "journal": "BMC cancer", "year": 2021, "abstract": "BACKGROUND: In recent years, gene expression-based analysis has been used for disease biomarker discovery, providing ways for better diagnosis, leading to improvement of clinical treatment efficacy. This study aimed to explore the role of miR-16-5p and ANLN in breast cancer (BC).\nMETHODS: Cohort datasets of BC were obtained from the Gene Expression Omnibus (GEO) and the Cancer Genome Atlas (TCGA) and analyzed by bioinformatics tools. qRT-PCR and western blotting were applied to validate ANLN and its protein expression. A dual-luciferase reporter assay was used to prove the regulatory relationship of miR-16-5p and ANLN. Finally, MTT, wound healing, Transwell invasion and flow cytometry analyses of the cell cycle and apoptosis were performed to assess cell proliferation, migration, invasion, cell cycle and apoptosis, respectively.\nRESULTS: A total of 195 differentially expressed genes (DEGs) and 50 overlapping microRNAs (miRNAs) were identified. Among these DEGs and miRNAs, ANLN, associated with poor overall survival in BC, overlapped in the GSE29431, GSE42568, TCGA and GEPIA2 databases. Moreover, ANLN was highly expressed, while miR-16-5p was lower in BC cells than in breast epithelial cells. Then, we confirmed that ANLN was directly targeted by miR-16-5p in BC cells. Over-expression of miR-16-5p and knock-down of ANLN remarkably inhibited cell proliferation and migration as well as cell invasion, arrested the cells in G2/M phase and induced apoptosis in BC cells.\nCONCLUSIONS: These findings suggest that miR-16-5p restrains proliferation, migration and invasion while affecting cell cycle and promotes apoptosis by regulating ANLN, thereby providing novel candidate biomarkers for the diagnosis and treatment of BC.", "claims": [{"id": "claim-method", "statement": "Through applying limma package, FDR < 0.05 and |logFC| > 1.5 [24, 25] were set as the screening threshold values to identify differentially expressed genes (DEGs) from GSE86374.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:34743685", "locator": "sections[3] \"Data sources and processing and identification of DEGs\" paragraph 2", "quote": "Through applying limma package, FDR < 0.05 and |logFC| > 1.5 [24, 25] were set as the screening threshold values to identify differentially expressed genes (DEGs) from GSE86374."}], "state": "complete"}, {"id": "claim-data-access", "statement": "The datasets of BC in this study were downloaded from the GEO and UCSC Xena (https://www.ncbi.nlm.nih.gov/geo/; https://xenabrowser.net/datapages/) databases.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:34743685", "locator": "sections[34] \"notes\" paragraph 1", "quote": "The datasets of BC in this study were downloaded from the GEO and UCSC Xena (https://www.ncbi.nlm.nih.gov/geo/; https://xenabrowser.net/datapages/) databases."}], "state": "complete"}, {"id": "claim-target-result", "statement": "These findings suggest that miR-16-5p restrains proliferation, migration and invasion while affecting cell cycle and promotes apoptosis by regulating ANLN, thereby providing novel candidate biomarkers for the diagnosis and treatment of BC.", "kind": "qualitative", "target": null, "evidence": [{"asset": "source:pmid:34743685", "locator": "sections[0] \"Abstract\" paragraph 4", "quote": "These findings suggest that miR-16-5p restrains proliferation, migration and invasion while affecting cell cycle and promotes apoptosis by regulating ANLN, thereby providing novel candidate biomarkers for the diagnosis and treatment of BC."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "complete", "objective": "These findings suggest that miR-16-5p restrains proliferation, migration and invasion while affecting cell cycle and promotes apoptosis by regulating ANLN, thereby providing novel candidate biomarkers for the diagnosis and treatment of BC.", "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:34743685"], "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:GSE29431"], "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:GSE29431"], "outputs": ["the replicated result and its comparison"], "instruments": [], "reagents": []}], "materials": [], "instruments": [], "reagents": [], "conditions": {}, "gaps": []}, "data": {"state": "partial", "assets": [{"id": "source:pmid:34743685", "role": "manuscript", "format": "xml", "media_type": "application/xml", "uri": "https://doi.org/10.1186/s12885-021-08914-1", "sha256": "sha256:e3d038503913d733ee535eab8bec2b076c98808a465d069c9f2793ffe743db27", "bytes": 34684, "access": "open", "state": "resolved", "reader": null, "reader_status": "reader_required", "identifier": "10.1186/s12885-021-08914-1", "version": null, "note": "Full article fetched from PMC; the checksum covers the rendered section body, not the original XML bytes.", "structure": {"pmcid": "PMC8574041", "source": "europe_pmc_fulltext", "coverage": "whole_body_without_references", "text_hash_scope": "rendered_section_body"}}, {"id": "data:geo:GSE29431", "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=GSE29431", "sha256": null, "bytes": null, "access": "open", "state": "resolved", "reader": null, "reader_status": "reader_required", "identifier": "GSE29431", "version": null, "note": "The datasets of BC in this study were downloaded from the GEO and UCSC Xena (https://www.ncbi.nlm.nih.gov/geo/; 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"identifier_resolution_only_no_download_no_licence_read", "downloaded": false, "licence_read": false}}], "gaps": ["asset:data:geo:GSE29431 not downloaded", "asset:data:geo:GSE29431 licence not read", "asset:data:geo:GSE42568 not downloaded", "asset:data:geo:GSE42568 licence not read", "asset:data:geo:GSE86374 not downloaded", "asset:data:geo:GSE86374 licence not read"]}, "analysis": {"state": "complete", "methods": [{"id": "method-1", "family": "differential_expression", "specific": "limma_voom", "label": "Differential expression analysis", "dsl_operations": ["normalise_counts", "differential_expression_fit", "fdr_bh"], "missing_operations": [], "expressible": true, "parameters": {}, "data_shapes": ["count_matrix_with_condition"], "evidence": {"score": 1.0, "band": "verbatim", "quote": "Through applying limma package, FDR < 0.05 and |logFC| > 1.5 [24, 25] were set as the screening threshold values to identify differentially expressed genes (DEGs) from GSE86374."}}], "targets": [], 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"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": ["GSE86374", "GSE29431", "GSE42568"], "resolved_accessions": ["GSE86374", "GSE29431", "GSE42568"], "statements": ["The datasets of BC in this study were downloaded from the GEO and UCSC Xena (https://www.ncbi.nlm.nih.gov/geo/; https://xenabrowser.net/datapages/) databases."], "basis": "The paper names GSE86374, GSE29431, GSE42568 in Gene Expression Omnibus (GEO) and the accession resolves in the repository. No dataset was downloaded and no licence was read."}}, "can_run": {"dsl_operations": ["normalise_counts", "differential_expression_fit", "fdr_bh"], "assets": ["source:pmid:34743685", "data:geo:GSE29431"], "steps": ["run_analysis"]}, "must_build": {"dsl_operations": [], "converters": [], "readers": [], "capabilities": [], "data_gaps": ["data:geo:GSE29431"]}, "gaps": []}, "review": {"state": "partial", "machine_critique": {"kind": "extraction_review", "reader": {"model": "MiniMax-M3.1-Flash", "provider": "minimax", "provider_family": "minimax", "read_at": "2026-11-01T17:06:18Z", "independence": "single_reader"}, "evidence_check": {"method": "verbatim", "availability": "verbatim"}, "calls": 5, "usage": {"prompt_tokens": 10868, "completion_tokens": 2531, "cached_tokens": 264}}, "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:34743685", "type": "paper", "label": "pmid:34743685"}, {"id": "source:pmid:34743685", "type": "manuscript", "label": "PMID 34743685 full text"}, {"id": "data:geo:GSE29431", "type": "dataset", "label": "GEO GSE29431"}], "activities": [{"id": "activity-reading-sci-2026-11-01", "type": "classification", "used": [{"id": "pmid:34743685", "type": "paper", "label": "pmid:34743685"}, {"id": "source:pmid:34743685", "type": "manuscript", "label": "PMID 34743685 full text"}, {"id": "data:geo:GSE29431", "type": "dataset", "label": "GEO GSE29431"}], "agent": "agent:corpus-assay-engine"}, {"id": "activity-read-34743685", "type": "extraction", "used": ["source:pmid:34743685"], "agent": "agent:minimax-reader"}, {"id": "activity-resolve-GSE29431", "type": "accession_resolution", "used": ["data:geo:GSE29431"], "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:34743685", "parent_hash": "sha256:352ffabd07ce5524d4a2f5bdeb677338a7e3a3ca7f780e797a0d8bc51092ba83", "fork_of": null, "generation": 4}, "gaps": ["asset:data:geo:GSE29431 licence not read", "asset:data:geo:GSE29431 not downloaded", "asset:data:geo:GSE42568 licence not read", "asset:data:geo:GSE42568 not downloaded", "asset:data:geo:GSE86374 licence not read", "asset:data:geo:GSE86374 not downloaded", "execution.not_run", "review.human_critique", "review.independent_reader", "review.machine_critique"], "assays": [{"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": ["nalyzed by bioinformatics tools. qRT-PCR and western blotting were applied to validate ANLN and its protei"], "order_basis": "paper_text_order:473", "id": "assay:1"}, {"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": ["low cytometry analyses of the cell cycle and apoptosis were performed to assess cell proliferation,", "eration, migration, invasion, cell cycle and apoptosis, respectively. RESULTS: A total of 195 diffe", "arrested the cells in G2/M phase and induced apoptosis in BC cells. CONCLUSIONS: These findings sug", "sion while affecting cell cycle and promotes apoptosis by regulating ANLN, thereby providing novel"], "order_basis": "paper_text_order:659", "id": "assay:2"}, {"role": "data_generation", "definition_id": "flow_cytometry", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": [", MTT, wound healing, Transwell invasion and flow cytometry analyses of the cell cycle and apoptosis wer", "factor at 37 °C in an incubator with 5% CO2. Flow cytometry analysis of cell cycle and apoptosis MCF-7 a", "odide stained cells for 30 min at 4 °C. A BD FACS caliber was used to measure cell cycle. BC c", "col. Finally, apoptosis was determined by BD FACS caliber. All experiments were independently"], "order_basis": "paper_text_order:702", "id": "assay:3"}, {"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": ["n BC cells. Over-expression of miR-16-5p and knock-down of ANLN remarkably inhibited cell proliferat", "ycling and cultured for 24 h. The cells were transfected with miR-16-5p mimics, si-ANLN and NC in eac", "ANLN and NC in each well for 24 h. Then, the transfected cells were collected. Precooled PBS and 70%", "ted as the same as cell cycle. After 24 h of transfection, cells were washed with PBS and collected in"], "order_basis": "paper_text_order:1344", "id": "assay:4"}, {"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": ["BC cell lines and HEK293T cells were mainly cultured in Dulbecco′s modified Eagle′s medium (DMEM) su", "bator with 5% CO2. MCF-10A cells were mainly cultured in Dulbecco′s modified Eagle′s medium/Ham′s F12", "Methods Cell culture The human cell lines MCF-7, T47D, MDA-MB-231, EMF-192A, SKBR-3 and MCF-", "tometry analysis of cell cycle and apoptosis MCF-7 and MDA-MB-231 cells (0.5 × 106 cells/well)"], "order_basis": "paper_text_order:1809", "id": "assay:5"}, {"role": "analysis", "definition_id": "differential_expression", "definition_version": "f2132e96ae6c7a4e", "definition_digest": null, "required_operations": ["normalise_counts", "differential_expression_fit", "fdr_bh"], "support_state": "supported", "source_hashes": {"dsl_catalogue": "f2132e96ae6c7a4e"}, "confidence": 1.0, "evidence": ["Through applying limma package, FDR < 0.05 and |logFC| > 1.5 [24, 25] were set as the screening threshold values to identify differentially expressed genes (DEGs) from GSE86374."], "order_basis": "consumes_earlier_output", "id": "assay:6"}]}