Molecular and cellular probes · 2025
Identification and experimental validation of biomarkers for acute myeloid leukemia based on single-cell RNA sequencing data.
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Yang Xin, Zhou Siyue, Chen Yuanju, Wang Ying, Cheng Yujing, Zhang Chan, Li Qi
Аннотация
BACKGROUND: Acute myeloid leukemia (AML) is a highly diverse malignant tumor at the molecular level with a high mortality rate. However, its underlying mechanisms remain poorly understood, and reliable biomarkers are lacking.
METHODS: AML-related biomarkers were identified through a comprehensive analysis of single-cell RNA sequencing (scRNA-seq), Mendelian randomization (MR), and gene regulatory networks. The biomarkers were then subjected to GSEA enrichment analysis and in vitro experimental validation.
RESULTS: scRNA-seq identified a total of 22,742 cells with detectable gene expression. Among the differentially expressed genes in the cells, 174 marker genes were screened out. MR analysis of marker genes showed that ITGB2, AIF1, CA2, CST7, and JCHAIN had a significant causal relationship with AML. Cumulative recovery curve analysis showed that AIF1, CST7, and ITGB2 were in the top motifs with the highest normalized enrichment scores. Therefore, they were recorded as biomarkers. Cellular experiments confirmed that AIF1, CST7, and ITGB2 promote cell proliferation and inhibit apoptosis.
CONCLUSION: This study identified AIF1, CST7, and ITGB2 as biomarkers for AML through scRNA-seq, MR, and transcription factor enrichment analysis. Additionally, in vitro experiments, including cell transfection, cell proliferation, and flow cytometry, validated the important role of these biomarkers in promoting the malignant phenotype of AML.
METHODS: AML-related biomarkers were identified through a comprehensive analysis of single-cell RNA sequencing (scRNA-seq), Mendelian randomization (MR), and gene regulatory networks. The biomarkers were then subjected to GSEA enrichment analysis and in vitro experimental validation.
RESULTS: scRNA-seq identified a total of 22,742 cells with detectable gene expression. Among the differentially expressed genes in the cells, 174 marker genes were screened out. MR analysis of marker genes showed that ITGB2, AIF1, CA2, CST7, and JCHAIN had a significant causal relationship with AML. Cumulative recovery curve analysis showed that AIF1, CST7, and ITGB2 were in the top motifs with the highest normalized enrichment scores. Therefore, they were recorded as biomarkers. Cellular experiments confirmed that AIF1, CST7, and ITGB2 promote cell proliferation and inhibit apoptosis.
CONCLUSION: This study identified AIF1, CST7, and ITGB2 as biomarkers for AML through scRNA-seq, MR, and transcription factor enrichment analysis. Additionally, in vitro experiments, including cell transfection, cell proliferation, and flow cytometry, validated the important role of these biomarkers in promoting the malignant phenotype of AML.