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To establish a radiation injury model using primary human lung fibroblasts and screen for key genes driving radiation-induced pulmonary fibrosis through transcriptome sequencing, thereby elucidating their potential mechanisms and providing new therapeutic targets and strategies for the clinical prevention and treatment of radiotherapy-induced lung injury.
Primary human lung fibroblasts were isolated by collagenase digestion and serially passaged. Cell purity was confirmed by flow cytometry prior to establishing an in vitro radiation model. Transcriptome analysis was performed based on qualified sequencing data. DESeq2 was used for differential expression analysis to identify upregulated genes, which were subsequently validated. Protein-protein interactions (PPI) among the differentially expressed gene sets were predicted using the String database and visualized using the igraph package. Finally, functional and pathway enrichment analyses of the core genes were performed via gene ontology (GO), Kyoto encyclopedia of genes and genomes (KEGG) and gene set enrichment analysis (GSEA) using the clusterProfiler software.
All samples met the quality control standards. Transcriptome analysis identified three significantly upregulated key differentially expressed genes: sulfatase 1 (SULF1), sulfatase 2 (SULF2), and oligodendrocyte myelin glycoprotein (OMG). These findings were consistently validated in a public dataset. PPI network analysis constructed with Cytoscape revealed that SULF1 occupied a central hub position within the network, exhibiting a strong connection with SULF2, and both directly interacted with OMG. GSEA indicated that SULF1/2 and OMG were significantly enriched in pathways related to DNA function, glutathione transferase activity, and neuroimmune regulation. These results suggested that SULF1/2 and OMG may function in radiation-induced lung injury by participating in processes such as DNA damage response, oxidative stress, and neuroimmune interactions.
SULF1, SULF2 and OMG are key molecules in human lung fibroblasts responding to radiation injury and may drive the progression of radiation-induced lung injury through multiple pathways. As potential therapeutic targets, they provide a novel theoretical basis for the clinical prevention and treatment of radiotherapy-induced lung injury.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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