MicroRNA-376b-3p targets RGS1 mRNA to inhibit proliferation, metastasis, and apoptosis in osteosarcoma.

{"sci_version": "1.2", "schema": "https://corpus.bntrg.co.uk/sci/1.0/sci.schema.json", "id": "pmid:34988161", "kind": "science-record", "revision": 5, "content_hash": "sha256:d8fc8692c8da5e4b3baea46044b8b2aac6df564d9e70b68d5e3cecf02edb7a9e", "created_at": "2026-11-01T17:36:42Z", "updated_at": "2026-11-01T22:48:10Z", "generator": {"name": "corpus-assay-engine", "run_id": "reading-sci-2026-11-01"}, "paper": {"state": "complete", "title": "MicroRNA-376b-3p targets RGS1 mRNA to inhibit proliferation, metastasis, and apoptosis in osteosarcoma.", "doi": "10.21037/atm-21-4949", "pmid": "34988161", "authors": ["Lei Zhang", "Meng Yao", "Weikang Ma", "Yongqing Jiang", "Wenbo Wang"], "journal": "Annals of translational medicine", "year": 2021, "abstract": "BACKGROUND: To investigate the role of microRNA-376b-3p (miR-376b-3p) and regulator of G protein signaling 1 (RGS1) in the proliferation, metastasis, and apoptosis of osteosarcoma.\nMETHODS: Differentially expressed genes (DEGs) between tumor and normal tissues from GSE14359 and GSE33382 in the cancer genome atlas (TCGA) dataset were analyzed with GEO2R online. Similarly, differentially expressed miRNAs from GSE70367 were also analyzed with GEO2R. The interaction between the differentially expressed miRNAs and the shared distal metastasis-related DEGs from the two datasets were analyzed using miRWalk and Cytoscape. RGS1 and miR-376-3p were chosen to verify the prediction. RGS1 stably expressing and silencing cells were established based on the MG63 and U2OS cell lines. The targeting of RGS1 with miR-376b-3p was confirmed with Starbase prediction and luciferase reporter assay. Cell proliferation, metastasis, and apoptosis were characterized in vitro and in xenograft mice.\nRESULTS: A total of 10 up-regulated and 8 down-regulated DEGs were characterized as shared metastasis-related DEGs for GSE14359 and GSE33382. Among these DEGs, RGS1 was targeted with miR-376b-3p, a predicted down-regulated miRNA in GSE70367. High expression of RGS1 predicted proliferation, invasion, metastases, and poor prognosis in osteosarcoma. Overexpression of RGS1 promoted proliferation, invasion, mobility, and stemness in MG63 and U2OS cells, while silencing of RGS1 had the opposite effect in both cell lines. High expression of RGS1 promoted tumor growth in xenograft nude mice. RGS1 was targeted with miR-376b-3p; the addition of miR-376b-3p down-regulated RGS1, and suppressed cell proliferation, invasion, and metastasis. Meanwhile, sponging of miR-376b-3p had the opposite effect. The suppressive effects of miR-376b-3p could be abolished with RGS1, as cell proliferation, stemness, metastasis, and invasion were all promoted with RGS1 co-transfection in both cell lines.\nCONCLUSIONS: Our study indicated that RGS1 is a tumor-promoting gene in osteosarcoma, which could be inhibited with miR-376b-3p.", "claims": [{"id": "claim-method", "statement": "For GEO2R analysis, Log2|Fold-Changes| (Log2 FC) >1 and P<0.05 was set as the screening criteria for DEGs.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:34988161", "locator": "sections[3] \"GEO2R online analysis of DEGs\" paragraph 2", "quote": "For GEO2R analysis, Log2|Fold-Changes| (Log2 FC) >1 and P<0.05 was set as the screening criteria for DEGs."}], "state": "complete"}, {"id": "claim-data-access", "statement": "Data Sharing Statement: Available at https://dx.doi.org/10.21037/atm-21-4949", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:34988161", "locator": "sections[28] \"(untitled)\" paragraph 1", "quote": "Data Sharing Statement: Available at https://dx.doi.org/10.21037/atm-21-4949"}], "state": "complete"}, {"id": "claim-target-result", "statement": "Among the shared metastasis-related DEGs, RGS1 was targeted with miR-376b-3p, a predicted down-regulated miRNA in GSE70367.", "kind": "qualitative", "target": null, "evidence": [{"asset": "source:pmid:34988161", "locator": "sections[0] \"Abstract\" paragraph 3", "quote": "Among these DEGs, RGS1 was targeted with miR-376b-3p, a predicted down-regulated miRNA in GSE70367."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "complete", "objective": "Among the shared metastasis-related DEGs, RGS1 was targeted with miR-376b-3p, a predicted down-regulated miRNA in GSE70367.", "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:34988161"], "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:GSE14359"], "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:GSE14359"], "outputs": ["the replicated result and its comparison"], "instruments": [], "reagents": []}], "materials": [], "instruments": [], "reagents": [], "conditions": {}, "gaps": []}, "data": {"state": "partial", "assets": [{"id": "source:pmid:34988161", "role": "manuscript", "format": "xml", "media_type": "application/xml", "uri": "https://doi.org/10.21037/atm-21-4949", "sha256": "sha256:7818d366b57bd39053b420f027391b3374d3d3bcbbd73c3b32dc17c70e97bc0a", "bytes": 36830, "access": "open", "state": "resolved", "reader": null, "reader_status": "reader_required", "identifier": "10.21037/atm-21-4949", "version": null, "note": "Full article fetched from PMC; 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METHODS: Differentially exp", "r assay. Cell proliferation, metastasis, and apoptosis were characterized in vitro and in xenograft"], "order_basis": "paper_text_order:32", "id": "assay:1"}, {"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": ["cells were established based on the MG63 and U2OS cell lines. The targeting of RGS1 with miR-3", "invasion, mobility, and stemness in MG63 and U2OS cells, while silencing of RGS1 had the oppos", "purchased from ATCC, Manassas, VA, USA) were cultured in Dulbecco’s modified Eagle’s medium containin", "fic, Inc., Waltham, MA, USA). The cells were cultured in a 37 °C humidified incubator (Thermo Fisher"], "order_basis": "paper_text_order:821", "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": 0.99, "evidence": ["and invasion were all promoted with RGS1 co-transfection in both cell lines. CONCLUSIONS: Our study i", "sinki (as revised in 2013). Cell culture and transfection MG63 and U2OS cell lines (purchased from ATC", "ic, Inc., Waltham, MA, USA) with 5% CO2. For transfection, a lentivirus-packaged RGS1 vector, short ha", "in Biotechology (Shanghai, China) (10). Cell transfection was applied according to the manufacturer’s"], "order_basis": "paper_text_order:2000", "id": "assay:3"}, {"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": ["enced cells were verified every 2 weeks with western blotting. The sequence of the shRNAs and mimics were"], "order_basis": "paper_text_order:3764", "id": "assay:4"}, {"role": "data_generation", "definition_id": "animal_model", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["tumor volume was recorded every 2 days. The mice were sacrificed 2 weeks later and the tumor volume and tumor", "rom Charles River Inc. (Beijing, China). The mice were raised in a SPF house, with a 12 h day/night", "tumor volume was recorded every 2 days. The mice were sacrificed 2 weeks later and the tumor volum", "nk skin at a density of 5×106 cells per nude mouse (n= five mice per group). The tumor volume w"], "order_basis": "paper_text_order:4302", "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": ["For GEO2R analysis, Log2|Fold-Changes| (Log2 FC) >1 and P<0.05 was set as the screening criteria for DEGs."], "order_basis": "consumes_earlier_output", "id": "assay:6"}]}