Identification and validation of HOXC6 as a diagnostic biomarker for Ewing sarcoma: insights from machine learning algorithms and in vitro experiments

IntroductionEarly diagnosis of Ewing sarcoma (ES) is critical for improving patient prognosis. However, the accurate diagnosis of ES remains challenging, underscoring the need for novel diagnostic biomarkers to enhance diagnostic precision and reliability. This study aimed to identify potential gene...

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Main Authors: Yonghua Pang, Jiahui Liang, Yakai Deng, Weinan Chen, Yunyan Shen, Jing Li, Xin Wang, Zhiyao Ren
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-04-01
Series:Frontiers in Immunology
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Online Access:https://www.frontiersin.org/articles/10.3389/fimmu.2025.1449355/full
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author Yonghua Pang
Jiahui Liang
Jiahui Liang
Yakai Deng
Weinan Chen
Yunyan Shen
Jing Li
Xin Wang
Zhiyao Ren
author_facet Yonghua Pang
Jiahui Liang
Jiahui Liang
Yakai Deng
Weinan Chen
Yunyan Shen
Jing Li
Xin Wang
Zhiyao Ren
author_sort Yonghua Pang
collection DOAJ
description IntroductionEarly diagnosis of Ewing sarcoma (ES) is critical for improving patient prognosis. However, the accurate diagnosis of ES remains challenging, underscoring the need for novel diagnostic biomarkers to enhance diagnostic precision and reliability. This study aimed to identify potential gene expression-based biomarkers for the diagnosis of ES.MethodsWe selected the GSE17679, GSE45544, and GSE68776 datasets from the Gene Expression Omnibus (GEO) database. After correcting for batch effects, we combined ES and normal tissue samples from the GSE17679 and GSE45544 datasets to create a combined cohort. Two-thirds of both the tumor and normal samples from the combined cohort were randomly selected for the training cohort, while the remaining one-third served as the internal validation cohort. Additionally, the GSE68776 dataset was used for external validation. To identify key diagnostic genes, we applied three machine learning algorithms: least absolute shrinkage and selection operator (LASSO), support vector machine recursive feature elimination (SVM-RFE), and random forest (RF).ResultsHOXC6 was identified as a key diagnostic biomarker for ES. It demonstrated strong diagnostic performance across all cohorts, with area under the curve (AUC) values of 0.956 (95% CI: 0.909−0.990) in the training cohort, 0.995 (95% CI: 0.977−1.000) in the internal validation cohort, and 0.966 (95% CI: 0.910−0.999) in the external validation cohort. Functional validation through HOXC6 knockdown in the RD-ES cell line revealed that its suppression significantly inhibited cell proliferation and migration. Furthermore, transcriptome sequencing suggested potential oncogenic mechanisms underlying HOXC6 function.DiscussionThese findings highlight HOXC6 as a promising diagnostic biomarker for ES, demonstrating robust performance across multiple datasets. Additionally, its functional role suggests potential as a therapeutic target.
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spelling doaj-art-aa633b8c2f7e4c5ea7ce7b07fb12d7c42025-08-20T03:07:28ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-04-011610.3389/fimmu.2025.14493551449355Identification and validation of HOXC6 as a diagnostic biomarker for Ewing sarcoma: insights from machine learning algorithms and in vitro experimentsYonghua Pang0Jiahui Liang1Jiahui Liang2Yakai Deng3Weinan Chen4Yunyan Shen5Jing Li6Xin Wang7Zhiyao Ren8Department of Orthopedics, The 904th Hospital of the Joint Logistics Support Force, People's Liberation Army of China, Wuxi, Jiangsu, ChinaDepartment of Breast Surgery, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, ChinaDepartment of General Surgery, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, ChinaDepartment of Orthopedics, The 904th Hospital of the Joint Logistics Support Force, People's Liberation Army of China, Wuxi, Jiangsu, ChinaDepartment of Orthopedics, The 904th Hospital of the Joint Logistics Support Force, People's Liberation Army of China, Wuxi, Jiangsu, ChinaDepartment of Orthopedics, The 904th Hospital of the Joint Logistics Support Force, People's Liberation Army of China, Wuxi, Jiangsu, ChinaDepartment of Orthopedics, Linyi People's Hospital, Linyi, Shandong, ChinaFaculty of Medicine and Health Sciences, Ghent University, Ghent, BelgiumFaculty of Medicine and Health Sciences, Ghent University, Ghent, BelgiumIntroductionEarly diagnosis of Ewing sarcoma (ES) is critical for improving patient prognosis. However, the accurate diagnosis of ES remains challenging, underscoring the need for novel diagnostic biomarkers to enhance diagnostic precision and reliability. This study aimed to identify potential gene expression-based biomarkers for the diagnosis of ES.MethodsWe selected the GSE17679, GSE45544, and GSE68776 datasets from the Gene Expression Omnibus (GEO) database. After correcting for batch effects, we combined ES and normal tissue samples from the GSE17679 and GSE45544 datasets to create a combined cohort. Two-thirds of both the tumor and normal samples from the combined cohort were randomly selected for the training cohort, while the remaining one-third served as the internal validation cohort. Additionally, the GSE68776 dataset was used for external validation. To identify key diagnostic genes, we applied three machine learning algorithms: least absolute shrinkage and selection operator (LASSO), support vector machine recursive feature elimination (SVM-RFE), and random forest (RF).ResultsHOXC6 was identified as a key diagnostic biomarker for ES. It demonstrated strong diagnostic performance across all cohorts, with area under the curve (AUC) values of 0.956 (95% CI: 0.909−0.990) in the training cohort, 0.995 (95% CI: 0.977−1.000) in the internal validation cohort, and 0.966 (95% CI: 0.910−0.999) in the external validation cohort. Functional validation through HOXC6 knockdown in the RD-ES cell line revealed that its suppression significantly inhibited cell proliferation and migration. Furthermore, transcriptome sequencing suggested potential oncogenic mechanisms underlying HOXC6 function.DiscussionThese findings highlight HOXC6 as a promising diagnostic biomarker for ES, demonstrating robust performance across multiple datasets. Additionally, its functional role suggests potential as a therapeutic target.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1449355/fullEwing sarcomabioinformaticsmachine learningHOXC6diagnostic biomarker
spellingShingle Yonghua Pang
Jiahui Liang
Jiahui Liang
Yakai Deng
Weinan Chen
Yunyan Shen
Jing Li
Xin Wang
Zhiyao Ren
Identification and validation of HOXC6 as a diagnostic biomarker for Ewing sarcoma: insights from machine learning algorithms and in vitro experiments
Frontiers in Immunology
Ewing sarcoma
bioinformatics
machine learning
HOXC6
diagnostic biomarker
title Identification and validation of HOXC6 as a diagnostic biomarker for Ewing sarcoma: insights from machine learning algorithms and in vitro experiments
title_full Identification and validation of HOXC6 as a diagnostic biomarker for Ewing sarcoma: insights from machine learning algorithms and in vitro experiments
title_fullStr Identification and validation of HOXC6 as a diagnostic biomarker for Ewing sarcoma: insights from machine learning algorithms and in vitro experiments
title_full_unstemmed Identification and validation of HOXC6 as a diagnostic biomarker for Ewing sarcoma: insights from machine learning algorithms and in vitro experiments
title_short Identification and validation of HOXC6 as a diagnostic biomarker for Ewing sarcoma: insights from machine learning algorithms and in vitro experiments
title_sort identification and validation of hoxc6 as a diagnostic biomarker for ewing sarcoma insights from machine learning algorithms and in vitro experiments
topic Ewing sarcoma
bioinformatics
machine learning
HOXC6
diagnostic biomarker
url https://www.frontiersin.org/articles/10.3389/fimmu.2025.1449355/full
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