HtrA3: a promising prognostic biomarker and therapeutic target for head and neck squamous cell carcinoma.

{"sci_version": "1.2", "schema": "https://corpus.bntrg.co.uk/sci/1.0/sci.schema.json", "id": "pmid:37842043", "kind": "science-record", "revision": 3, "content_hash": "sha256:897b2235b4637d1a809fcc8f6f5e8d6a869783da82d686fd518853ad8e9f23aa", "created_at": "2026-11-01T19:02:42Z", "updated_at": "2026-11-01T22:10:28Z", "generator": {"name": "corpus-assay-engine", "run_id": "reading-sci-2026-11-01"}, "paper": {"state": "complete", "title": "HtrA3: a promising prognostic biomarker and therapeutic target for head and neck squamous cell carcinoma.", "doi": "10.7717/peerj.16237", "pmid": "37842043", "authors": ["Yan Chen", "Jianfeng Yang", "Hangbin Jin", "Weiwei Wen", "Ying Xu", "Xiaofeng Zhang", "Yu Wang"], "journal": "PeerJ", "year": 2023, "abstract": "Objective: The dysregulation of the human high-temperature requirement A (HtrA) family of serine proteases is associated with many malignancies. However, there are few reports on HtrAs in head and neck squamous cell carcinoma (HNSCC). The aim of this study was to investigate the expression, prognostic value, and biological functions of HtrAs in HNSCC.\nMethods: The RNA-sequencing data and clinical data of HNSCC were downloaded from The Cancer Genome Atlas (TCGA) database. The GSE30784 and GSE31056 datasets from the Gene Expression Omnibus (GEO) database were used for further verification. This study explored the differential expression of HtrAs and assessed their potential impact on the prognosis of HNSCC patients using a survival module. Correlations between clinical characteristics and HtrA expression levels were then explored using a Wilcoxon rank sum test. A Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Set Enrichment Analysis (GSEA) were performed using \"clusterProfile\" in the R software. A Pearson/Spearman correlation test was applied to analyze the relationship between HtrAs and immune infiltration level/checkpoint genes. Validation of HtrA expression levels were carried out by RT-PCR and western blot in human squamous carcinoma cell lines (Fadu and Cal-27) and human non-tumorigenic bronchial epithelium cells (BEAS-2B). Finally, through cell transfection, CCK-8, Ki-67 immunofluorescence, and flow cytometry assays, the effect of HtrA3 knockdown on the malignant biological behavior of HNSCC cells was explored.\nResults: The gene expression levels of HtrAs were significantly upregulated and associated with patient age, TNM stage, clinical stage, and TP53 mutation status in the TCGA-HNSCC cohort. High expressions of HtrA1/3 were associated with shorter overall survival, shorter progress-free interval, and lower disease-specific survival in HNSCC. A nomogram for HtrAs was constructed and validated. HtrA-related genes were significantly enriched in the immune response and cell apoptosis pathway. In addition, the expression of HtrAs showed significant correlations with B cells, M cells, DC cell infiltration, and immune infiltration checkpoint (CD276, TNFRSF14). Validation of HtrA expression was carried out by RT-PCR and western blot. Results of in vitro experiments indicated that HtrA3 gene knockdown inhibits the proliferation of FaDu and Cal-27 cells while concurrently promoting apoptosis.\nConclusions: HtrA3 shows significant potential as both a prognostic marker and a promising therapeutic target for HNSCC, highlighting its relevance and importance in future research and potential clinical applications.", "claims": [{"id": "claim-method", "statement": "A log rank test was used for the survival analysis.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:37842043", "locator": "sections[15] \"Statistical analysis\" paragraph 2", "quote": "A log rank test was used for the survival analysis."}], "state": "complete"}, {"id": "claim-data-access", "statement": "The following information was supplied regarding data availability:\nThe data is available at NCBI GEO: GSE30784, GSE31056.\nThe raw measurements are available in the Supplemental Files.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:37842043", "locator": "sections[38] \"Data Availability Statement\" paragraphs 1-4", "quote": "The following information was supplied regarding data availability:\nThe data is available at NCBI GEO: GSE30784, GSE31056.\nThe raw measurements are available in the Supplemental Files."}], "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:37842043"], "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. 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Methods: The RNA-sequencing data and clinical data of HNSCC were downloa"], "order_basis": "paper_text_order:428", "id": "assay:1"}, {"role": "data_generation", "definition_id": "western_blot", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["ession levels were carried out by RT-PCR and western blot in human squamous carcinoma cell lines (Fadu", "trA expression was carried out by RT-PCR and western blot. Results of in vitro experiments indicated t", "analyze correlation. Band intensities on the western blot were quantified using ImageJ software (versi"], "order_basis": "paper_text_order:1306", "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": ["elium cells (BEAS-2B). Finally, through cell transfection, CCK-8, Ki-67 immunofluorescence, and flow c", "d flow cytometry assays, the effect of HtrA3 knockdown on the malignant biological behavior of HNSC", "vitro experiments indicated that HtrA3 gene knockdown inhibits the proliferation of FaDu and Cal-2", "ta file. Supplemental Information 7Raw data: Transfection qPCR10.7717/peerj.16237/supp-7Click here for"], "order_basis": "paper_text_order:1462", "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": 0.99, "evidence": ["tly enriched in the immune response and cell apoptosis pathway. In addition, the expression of HtrA", "nd Cal-27 cells while concurrently promoting apoptosis. Conclusions: HtrA3 shows significant potent", "EAS-2B). Finally, through cell transfection, CCK-8, Ki-67 immunofluorescence, and flow cytometr"], "order_basis": "paper_text_order:1476", "id": "assay:4"}, {"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": ["lly, through cell transfection, CCK-8, Ki-67 immunofluorescence, and flow cytometry assays, the effect of Ht", ". Finally, through cell transfection, CCK-8, Ki-67 immunofluorescence, and flow cytometry assay", "ta file. Supplemental Information 9Raw data: Ki67-FAUD10.7717/peerj.16237/supp-9Click here for", "a file. Supplemental Information 10Raw data: Ki67-CAL2710.7717/peerj.16237/supp-10Click here f"], "order_basis": "paper_text_order:1483", "id": "assay:5"}, {"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": ["ection, CCK-8, Ki-67 immunofluorescence, and flow cytometry assays, the effect of HtrA3 knockdown on the"], "order_basis": "paper_text_order:1513", "id": "assay:6"}, {"role": "data_generation", "definition_id": "qpcr", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.93, "evidence": ["ta file. Supplemental Information 5Raw data: qPCR Validation10.7717/peerj.16237/supp-5Click he"], "order_basis": "paper_text_order:4288", "id": "assay:7"}, {"role": "analysis", "definition_id": "survival_analysis", "definition_version": "f2132e96ae6c7a4e", "definition_digest": null, "required_operations": ["logrank_test"], "support_state": "supported", "source_hashes": {"dsl_catalogue": "f2132e96ae6c7a4e"}, "confidence": 1.0, "evidence": ["A log rank test was used for the survival analysis."], "order_basis": "consumes_earlier_output", "id": "assay:8"}]}