骨肉瘤是最常见的原发性恶性骨肿瘤。由于化疗药物对肿瘤细胞的选择性和敏感性不足，加上使用大剂量药物，化疗药物常常具有全身性毒性。利用现代测序技术筛选大量肿瘤样本中的肿瘤标志物是筛选高度特异和选择性抗肿瘤药物的常用方法。本研究旨在利用最新报告的肿瘤源性癌基因诱导的复制应激（ORS）基因表达特征，识别潜在的生物标志物，并在不同的药物数据库中筛选潜在的抗骨肉瘤药物。在本研究中，我们在TARGET数据库中获得了89个与骨肉瘤相关的样本，所有样本均包括生存信息。根据六个已报告的ORS基因标记（NAT10/DDX27/ZNF48/C8ORF33/MOCS3/MPP6）的中位表达水平，我们将89个骨肉瘤基因表达数据集分为高表达组和低表达组，然后进行差异表达基因(DEG)分析。六组差异表达基因的共存基因被用作骨肉瘤的复制应激相关基因(RSGs)。然后，使用LASSO回归、Cox风险比例回归预测模型和十折交叉验证测试筛选关键的RSGs。从Gene Expression Omnibus (GEO)数据库中收集的GSE21257数据集用于验证预测模型。最终选定的关键RSGs用于在L1000PWD和DGIdb数据库中挖掘潜在药物。在经过预测模型的进一步验证后，我们确定了与骨肉瘤中的ORS有关的七个基因作为关键的RSGs，包括转录因子7类似2(TCF7L2)，溶质载体家族27成员4(SLC27A4)，前蛋白酶酶/切割酶5(PCSK5)，核仁蛋白6(NOL6)，螺旋螺旋螺旋螺旋结构域蛋白4(CHCHD4)，真核生物翻译起始因子3亚单位B(EIF3B)和细胞色素C氧化酶1的合成(SCO1)。然后，我们在两个药物数据库中筛选了这七个关键的RSGs，并发现了六种潜在的抗骨肉瘤药物(DGIdb数据库：雷帕格列奈、他克莫司、雷帕霉素、环孢素和氢氯噻嗪；L1000PWD数据库：小分子VU-0365117-1)。七个RSGs (TCF7L2, SLC27A4, PCSK5, NOL6, CHCHD4, EIF3B, 和SCO1) 可能与骨肉瘤中的ORS基因特征相关。雷帕格列奈、他克莫司、雷帕霉素、环孢素、氢氯噻嗪和小分子VU-0365117-1是骨肉瘤的潜在治疗药物。© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.

Osteosarcoma is the most common type of primary malignant bone tumor. Due to the lack of selectivity and sensitivity of chemotherapy drugs to tumor cells, coupled with the use of large doses, chemotherapy drugs often have systemic toxicity. The use of modern sequencing technology to screen tumor markers in a large number of tumor samples is a common method for screening highly specific and selective anti-tumor drugs. This study aims to identify potential biomarkers using the latest reported gene expression signatures of oncogene-induced replication stress (ORS) in aggressive cancers, and potential anti-osteosarcoma drugs were screened in different drug databases. In this study, we obtained 89 osteosarcoma-related samples in the TARGET database, all of which included survival information. According to the median expression of each of six reported ORS gene markers (NAT10/DDX27/ZNF48/C8ORF33/MOCS3/MPP6), we divided 89 osteosarcoma gene expression datasets into a high expression group and a low expression group and then performed a differentially expressed gene (DEG) analysis. The coexisting genes of 6 groups of DEGs were used as replication stress-related genes (RSGs) of osteosarcoma. Then, key RSGs were screened using LASSO regression, a Cox risk proportional regression prognostic model and a tenfold cross-validation test. GSE21257 datasets collected from the Gene Expression Omnibus (GEO) database were used to verify the prognostic model. The final key RSGs selected were used in the L1000PWD and DGIdb databases to mine potential drugs. After further validation by the prognostic model, we identified seven genes associated with ORS in osteosarcoma as key RSGs, including transcription factor 7 like 2 (TCF7L2), solute carrier family 27 member 4 (SLC27A4), proprotein convertase subtilisin/kexin type 5 (PCSK5), nucleolar protein 6 (NOL6), coiled-coil-coil-coil-coil-helix domain containing 4 (CHCHD4), eukaryotic translation initiation factor 3 subunit B (EIF3B), and synthesis of cytochrome C oxidase 1 (SCO1). Then, we screened the seven key RSGs in two drug databases and found six potential anti-osteosarcoma drugs (D GIdb database: repaglinide, tacrolimus, sirolimus, cyclosporine, and hydrochlorothiazide; L1000PWD database: the small molecule VU-0365117-1). Seven RSGs (TCF7L2, SLC27A4, PCSK5, NOL6, CHCHD4, EIF3B, and SCO1) may be associated with the ORS gene signatures in osteosarcoma. Repaglinide, tacrolimus, sirolimus, cyclosporine, hydrochlorothiazide and the small molecule VU-0365117-1 are potential therapeutic drugs for osteosarcoma.© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.