MiR-96-5p Suppresses Progression of Arsenite-Induced Human Keratinocyte Proliferation and Malignant Transformation by Targeting Denticleless E3 Ubiquitin Protein Ligase Homolog.

{"sci_version": "1.2", "schema": "https://corpus.bntrg.co.uk/sci/1.0/sci.schema.json", "id": "pmid:38133379", "kind": "science-record", "revision": 4, "content_hash": "sha256:316fdb0db744545705d2cdf48ca739b4c2932f8f50b8ce6853766f7a95fe5776", "created_at": "2026-11-01T19:20:03Z", "updated_at": "2026-11-01T22:53:20Z", "generator": {"name": "corpus-assay-engine", "run_id": "reading-sci-2026-11-01"}, "paper": {"state": "complete", "title": "MiR-96-5p Suppresses Progression of Arsenite-Induced Human Keratinocyte Proliferation and Malignant Transformation by Targeting Denticleless E3 Ubiquitin Protein Ligase Homolog.", "doi": "10.3390/toxics11120978", "pmid": "38133379", "authors": ["Yan Li", "Qiaoshi Zhao", "Jinyin Yao", "Chunpeng Lv", "Yanhui Gao", "Dianjun Sun", "Yanmei Yang"], "journal": "Toxics", "year": 2023, "abstract": "Long-term exposure to arsenic has been linked to a variety of cancers, among which skin cancer is the most prevalent form. However, the mechanism underlying arsenic carcinogenesis is unclear, and there is still limited information on the role of miRNAs in arsenic-induced skin cancer. This study aims to explore the role of miR-96-5p in the arsenite-induced proliferation and malignant transformation of human HaCaT keratinocytes. The GEO database (accession numbers GSE97303, GSE97305, and GSE97306) was used to extract mRNA and miRNA expression profiles of HaCaT cells treated with or without 0.1 μmol/L sodium arsenite for 3 and 7 weeks. In this paper, according to the CCK8 assay result, HaCaT cells exposed to 0.1 μmol/L sodium arsenite for 48 h were finalized. CCK8, MTT, EdU incorporation, and colony formation assays were used to determine the viability and proliferation of HaCaT cells and transformed HaCaT (T-HaCaT) cells. The subcellular localization and relative expression levels of DTL, as well as miR-96-5p in HaCaT cells induced by arsenite, were determined via immunofluorescence, RT-qPCR, and Western blot. Dual-luciferase reporter assay was performed to identify miR-96-5p bound directly to DTL. Transfection of miR-96-5p mimics or DTL siRNA was conducted to verify the arsenite-induced viability of HaCaT cells and T-HaCaT cells. T-HaCaT cells and nude mice were used to construct arsenite-induced malignant transformation and an in vivo xenograft model to demonstrate the over-expressed effect of miR-96-5p. The results showed that DTL was the target gene of miR-96-5p. Meanwhile, we also found that 0.1 μmol/L sodium arsenite upregulated DTL by decreasing the miR-96-5p level, leading to the proliferation and malignant transformation of HaCaT cells. MiR-96-5p agomir treatment slowed the growth of transplanted HaCaT cells transformed by arsenite in a manner associated with DTL downregulation in the nude mice xenograft model. Taken together, we confirmed that miR-96-5p, as a potent regulator of DTL, suppressed arsenite-induced HaCaT cell proliferation and malignant transformation, which might provide a novel therapeutic target for the treatment of arsenic-induced skin cancer.", "claims": [{"id": "claim-method", "statement": "The statistical significance between the two groups was assessed using Student’s 2-tailed t-test. One-way ANOVA followed by a Bonferroni-Dunn test was used for the comparison of more than two groups.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:38133379", "locator": "sections[17] \"2.15. Statistical Analysis\" paragraph 2", "quote": "The statistical significance between the two groups was assessed using Student’s 2-tailed t-test. One-way ANOVA followed by a Bonferroni-Dunn test was used for the comparison of more than two groups."}], "state": "complete"}, {"id": "claim-data-access", "statement": "The data presented in this study are available on request from the corresponding author.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:38133379", "locator": "sections[29] \"Data Availability Statement\" paragraph 2", "quote": "The data presented in this study are available on request from the corresponding author."}], "state": "complete"}, {"id": "claim-target-result", "statement": "Arsenite upregulates DTL by reducing miR-96-5p, promoting HaCaT-cell proliferation and malignant transformation; increasing miR-96-5p suppresses these effects by downregulating DTL.", "kind": "quantitative", "target": {"value": 29.0, "unit": "per cent", "tolerance": null, "source": "As shown in Figure 5D, when miR-96-5p mimics increased miR-96-5p level in HaCaT cells, the level of DTL mRNA in the miR-96-5p mimic-transfected cells reduced 29% (p < 0.01, Figure 5E), compared with cells transfected with the scramble, supporting the idea that miR-96-5p is capable of targeting the DTL 3′UTR to prevent DTL expression."}, "evidence": [{"asset": "source:pmid:38133379", "locator": "sections[0] \"Abstract\" paragraph 1", "quote": "The results showed that DTL was the target gene of miR-96-5p. Meanwhile, we also found that 0.1 μmol/L sodium arsenite upregulated DTL by decreasing the miR-96-5p level, leading to the proliferation and malignant transformation of HaCaT cells."}, {"asset": "source:pmid:38133379", "locator": "sections[22] \"3.4. MiR-96-5p, a Negative Regulator of DTL, Mediated the Proliferation of HaCaT Cells Induced by Arsenite\" paragraph 5", "quote": "As shown in Figure 5D, when miR-96-5p mimics increased miR-96-5p level in HaCaT cells, the level of DTL mRNA in the miR-96-5p mimic-transfected cells reduced 29% (p < 0.01, Figure 5E), compared with cells transfected with the scramble, supporting the idea that miR-96-5p is capable of targeting the DTL 3′UTR to prevent DTL expression."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "complete", "objective": "Arsenite upregulates DTL by reducing miR-96-5p, promoting HaCaT-cell proliferation and malignant transformation; increasing miR-96-5p suppresses these effects by downregulating DTL.", "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:38133379"], "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:GSE97303"], "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:GSE97303"], "outputs": ["the replicated result and its comparison"], "instruments": [], "reagents": []}], "materials": [], "instruments": [], "reagents": [], "conditions": {}, "gaps": []}, "data": {"state": "partial", "assets": [{"id": "source:pmid:38133379", "role": "manuscript", "format": "xml", "media_type": "application/xml", "uri": "https://doi.org/10.3390/toxics11120978", "sha256": "sha256:bff26ad26c1cdd270517d1ca8dae7dd31b98229d67404b6655d0070730aa4408", "bytes": 60199, "access": "open", "state": "resolved", "reader": null, "reader_status": "reader_required", "identifier": "10.3390/toxics11120978", "version": null, "note": "Full article fetched from PMC; the checksum covers the rendered section body, not the original XML bytes.", "structure": {"pmcid": "PMC10747408", "source": "europe_pmc_fulltext", "coverage": "whole_body", "text_hash_scope": "rendered_section_body"}}, {"id": "data:geo:GSE97303", "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=GSE97303", "sha256": null, "bytes": null, "access": "open", "state": "resolved", "reader": null, "reader_status": "reader_required", "identifier": "GSE97303", "version": null, "note": "The data presented in this study are available on request from the corresponding author.", "structure": {"repository": "GEO", "access_route": "GEO accession URL", "access_statement_verbatim": "The data presented in this study are available on request from the corresponding author.", "access_statement_kind": "data_availability_statement", "sample_accessions": ["GSM2561403", "GSM2561404", "GSM2561405", "GSM2561406", "GSM2561407", "GSM2561408", "GSM2561409", "GSM2561410", "GSM2561411", "GSM2561412", "GSM2561413", "GSM2561414", "GSM2561415", "GSM2561416", "GSM2561417", "GSM2561418", "GSM2561419", "GSM2561420"], "sample_count": 18, "group_sizes": null, "group_sizes_basis": null, "platform": "15207", "taxon": "Homo sapiens", "title": "Differential miRNA and mRNA Expression in Immortalized Human Keratinocytes (HaCaT) after Low Arsenic Exposure [mRNA]", "resolution_checked_at": "2026-11-01T19:09:46Z", "resolution_check": "identifier_resolution_only_no_download_no_licence_read", "downloaded": false, "licence_read": false}}, {"id": "data:geo:GSE97305", "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=GSE97305", "sha256": null, "bytes": null, "access": "open", "state": "resolved", "reader": null, "reader_status": "reader_required", "identifier": "GSE97305", "version": null, "note": "The data presented in this study are available on request from the corresponding author.", "structure": {"repository": "GEO", "access_route": "GEO accession URL", "access_statement_verbatim": "The data presented in this study are available on request from the corresponding author.", "access_statement_kind": "data_availability_statement", "sample_accessions": ["GSM2561453", "GSM2561454", "GSM2561455", "GSM2561456", "GSM2561457", "GSM2561458", "GSM2561459", "GSM2561460", "GSM2561461", "GSM2561462", "GSM2561463", "GSM2561464", "GSM2561465", "GSM2561466", "GSM2561467", "GSM2561468", "GSM2561469", "GSM2561470"], "sample_count": 18, "group_sizes": null, "group_sizes_basis": null, "platform": "19117", "taxon": "synthetic construct; Homo sapiens", "title": "Differential miRNA and mRNA Expression in Immortalized Human Keratinocytes (HaCaT) after Low Arsenic Exposure [miRNA]", "resolution_checked_at": "2026-11-01T19:09:46Z", "resolution_check": "identifier_resolution_only_no_download_no_licence_read", "downloaded": false, "licence_read": false}}, {"id": "data:geo:GSE97306", "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=GSE97306", "sha256": null, "bytes": null, "access": "open", "state": "resolved", "reader": null, "reader_status": "reader_required", "identifier": "GSE97306", "version": null, "note": "The data presented in this study are available on request from the corresponding author.", "structure": {"repository": "GEO", "access_route": "GEO accession URL", "access_statement_verbatim": "The data presented in this study are available on request from the corresponding author.", "access_statement_kind": "data_availability_statement", "sample_accessions": ["GSM2561403", "GSM2561404", "GSM2561405", "GSM2561406", "GSM2561407", "GSM2561408", "GSM2561409", "GSM2561410", "GSM2561411", "GSM2561412", "GSM2561413", "GSM2561414", "GSM2561415", "GSM2561416", "GSM2561417", "GSM2561418", "GSM2561419", "GSM2561420", "GSM2561453", "GSM2561454", "GSM2561455", "GSM2561456", "GSM2561457", "GSM2561458", "GSM2561459", "GSM2561460", "GSM2561461", "GSM2561462", "GSM2561463", "GSM2561464", "GSM2561465", "GSM2561466", "GSM2561467", "GSM2561468", "GSM2561469", "GSM2561470"], "sample_count": 36, "group_sizes": null, "group_sizes_basis": null, "platform": "19117;15207", "taxon": "Homo sapiens; synthetic construct", "title": "Differential miRNA and mRNA Expression in Immortalized Human Keratinocytes (HaCaT) after Low Arsenic Exposure", "resolution_checked_at": "2026-11-01T19:09:46Z", "resolution_check": "identifier_resolution_only_no_download_no_licence_read", "downloaded": false, "licence_read": false}}], "gaps": ["asset:data:geo:GSE97303 not downloaded", "asset:data:geo:GSE97303 licence not read", "asset:data:geo:GSE97305 not downloaded", "asset:data:geo:GSE97305 licence not read", "asset:data:geo:GSE97306 not downloaded", "asset:data:geo:GSE97306 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 statistical significance between the two groups was assessed using Student’s 2-tailed t-test. One-way ANOVA followed by a Bonferroni-Dunn test was used for the comparison of more than two groups."}}], "targets": [{"id": "target-published-estimate", "claim_id": "claim-target-result", "operation": "bootstrap_mean_difference", "expected_value": 29.0, "unit": "per cent", "tolerance": null, "state": "complete", "note": "The published estimate as the reader quoted it from the paper. No tolerance is published because the paper states none."}, {"id": "target-published-estimate-p-value", "claim_id": "claim-target-result", "operation": "bootstrap_mean_difference", "expected_value": 0.01, "unit": "p-value", "tolerance": null, "state": "complete", "note": "The threshold the paper applied to its own result, as the paper states it. A run that reproduces the effect is judged against this threshold."}], "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": ["GSE97303", "GSE97305", "GSE97306"], "resolved_accessions": ["GSE97303", "GSE97305", "GSE97306"], "statements": ["The data presented in this study are available on request from the corresponding author."], "basis": "The paper names GSE97303, GSE97305, GSE97306 in GEO database 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:38133379", "data:geo:GSE97303"], "steps": ["run_analysis"]}, "must_build": {"dsl_operations": [], "converters": [], "readers": [], "capabilities": [], "data_gaps": ["data:geo:GSE97303"]}, "gaps": []}, "review": {"state": "partial", "machine_critique": {"kind": "extraction_review", "reader": {"model": "MiniMax-M3.1-Flash", "provider": "minimax", "provider_family": "minimax", "read_at": "2026-11-01T19:03:14Z", "independence": "single_reader"}, "evidence_check": {"method": "verbatim", "availability": "verbatim"}, "calls": 5, "usage": {"prompt_tokens": 18912, "completion_tokens": 1610, "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:38133379", "type": "paper", "label": "pmid:38133379"}, {"id": "source:pmid:38133379", "type": "manuscript", "label": "PMID 38133379 full text"}, {"id": "data:geo:GSE97303", "type": "dataset", "label": "GEO GSE97303"}], "activities": [{"id": "activity-reading-sci-2026-11-01", "type": "classification", "used": [{"id": "pmid:38133379", "type": "paper", "label": "pmid:38133379"}, {"id": "source:pmid:38133379", "type": "manuscript", "label": "PMID 38133379 full text"}, {"id": "data:geo:GSE97303", "type": "dataset", "label": "GEO GSE97303"}], "agent": "agent:corpus-assay-engine"}, {"id": "activity-read-38133379", "type": "extraction", "used": ["source:pmid:38133379"], "agent": "agent:minimax-reader"}, {"id": "activity-resolve-GSE97303", "type": "accession_resolution", "used": ["data:geo:GSE97303"], "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:38133379", "parent_hash": "sha256:40e9ba3af3af9d4f7175d551925f5410888c6e9d45930322383097799c026a5f", "fork_of": null, "generation": 3}, "gaps": ["asset:data:geo:GSE97303 licence not read", "asset:data:geo:GSE97303 not downloaded", "asset:data:geo:GSE97305 licence not read", "asset:data:geo:GSE97305 not downloaded", "asset:data:geo:GSE97306 licence not read", "asset:data:geo:GSE97306 not downloaded", "execution.not_run", "review.human_critique", "review.independent_reader", "review.machine_critique"], "assays": [{"role": "data_generation", "definition_id": "clinical_trial", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.93, "evidence": ["and miRNA expression profiles of HaCaT cells treated with or without 0.1 μmol/L sodium arsenite for 3 and 7 weeks"], "order_basis": "paper_text_order:704", "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": ["and 7 weeks. In this paper, according to the CCK8 assay result, HaCaT cells exposed to 0.1 μmo", "l/L sodium arsenite for 48 h were finalized. CCK8, MTT, EdU incorporation, and colony formatio", "dium arsenite for 48 h were finalized. CCK8, MTT, EdU incorporation, and colony formation ass"], "order_basis": "paper_text_order:806", "id": "assay:2"}, {"role": "data_generation", "definition_id": "ihc", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.93, "evidence": ["lls induced by arsenite, were determined via immunofluorescence, RT-qPCR, and Western blot. Dual-luciferase"], "order_basis": "paper_text_order:1212", "id": "assay:3"}, {"role": "data_generation", "definition_id": "qpcr", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["ite, were determined via immunofluorescence, RT-qPCR, and Western blot. Dual-luciferase reporter", ", were determined via immunofluorescence, RT-qPCR, and Western blot. Dual-luciferase reporter"], "order_basis": "paper_text_order:1232", "id": "assay:4"}, {"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": ["ermined via immunofluorescence, RT-qPCR, and Western blot. Dual-luciferase reporter assay was performe"], "order_basis": "paper_text_order:1245", "id": "assay:5"}, {"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": ["to identify miR-96-5p bound directly to DTL. Transfection of miR-96-5p mimics or DTL siRNA was conduct", "DTL. Transfection of miR-96-5p mimics or DTL siRNA was conducted to verify the arsenite-induced"], "order_basis": "paper_text_order:1349", "id": "assay:6"}, {"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": ["ls and T-HaCaT cells. T-HaCaT cells and nude mice were used to construct arsenite-induced malignant", "nite-induced malignant transformation and an in vivo xenograft model to demonstrate the over-expr"], "order_basis": "paper_text_order:1507", "id": "assay:7"}, {"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": ["hnology Co., Ltd. (Wuhan, China). Cells were cultured in a DMEM medium (HyClone, Logan, UT, USA) supp"], "order_basis": "paper_text_order:2562", "id": "assay:8"}, {"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 statistical significance between the two groups was assessed using Student’s 2-tailed t-test. One-way ANOVA followed by a Bonferroni-Dunn test was used for the comparison of more than two groups."], "order_basis": "consumes_earlier_output", "id": "assay:9"}]}