骨肉瘤在恶性骨肿瘤中预后最差，且缺乏有效的生物标志物。我们的研究旨在探索m6A相关和免疫相关的生物标志物。我们从多个公共数据库中下载了骨肉瘤和健康对照组的基因表达谱，并利用生物信息学对其基于m6A的基因表达进行肿瘤分型。随后，我们利用最小绝对收缩和选择算子以及多元Cox回归分析构建了骨肉瘤的预后模型，并使用CIBERSORTx算法计算其免疫细胞组成。我们还对这两个基因进行了药物敏感性分析。最后，我们使用免疫组化验证了分析结果。我们还在体外实验中通过qRT-PCR检测了RBM15基因。我们收集了1738例骨肉瘤患者和24,344例非骨肉瘤患者的常规血液数据，并使用两个独立样本t检验验证了CIBERSORTx分析免疫细胞差异的准确性。基于m6A基因表达的肿瘤分型分析是最可靠的。基于RNA结合蛋白15(RBM15)和YTDC1构成的两个基因的预后模型高危组患者的生存率显著低于低危组患者(P < 0.05)。CIBERSORTx免疫细胞组分分析表明，RBM15与gama delta T细胞呈负相关，与活化自然杀伤细胞呈正相关。药物敏感性分析显示，这两个基因与多种药物呈现不同程度的相关性。免疫组化结果表明，这两个基因在骨肉瘤中的表达显著高于旁肿组织。qRT-PCR实验结果显示，在骨肉瘤细胞系和对照细胞系中，RBM15的表达显著高于对照组。绝对淋巴细胞值、淋巴细胞百分比、红细胞压积和红细胞计数在骨肉瘤组中均低于对照组(P < 0.001)。RBM15和YTHDC1可作为与m6A相关的骨肉瘤潜在预后生物标志物。©2023版权所有。

Osteosarcoma has the worst prognosis among malignant bone tumors, and effective biomarkers are lacking. Our study aims to explore m6A-related and immune-related biomarkers. Gene expression profiles of osteosarcoma and healthy controls were downloaded from multiple public databases, and their m6A-based gene expression was utilized for tumor typing using bioinformatics. Subsequently, a prognostic model for osteosarcoma was constructed using the least absolute shrinkage and selection operator and multivariate Cox regression analysis, and its immune cell composition was calculated using the CIBERSORTx algorithm. We also performed drug sensitivity analysis for these two genes. Finally, analysis was validated using immunohistochemistry. We also examined the RBM15 gene by qRT-PCR in an in vitro experiment. We collected routine blood data from 1738 patients diagnosed with osteosarcoma and 24,344 non-osteosarcoma patients and used two independent sample t tests to verify the accuracy of the CIBERSORTx analysis for immune cell differences. The analysis based on m6A gene expression tumor typing was most reliable using the two typing methods. The prognostic model based on the two genes constituting RNA-binding motif protein 15 (RBM15) and YTDC1 had a much lower survival rate for patients in the high-risk group than those in the low-risk group (P < 0.05). CIBERSORTx immune cell component analysis demonstrated that RBM15 showed a negative and positive correlation with T cells gamma delta and activated natural killer cells, respectively. Drug sensitivity analysis showed that these two genes showed varying degrees of correlation with multiple drugs. The results of immunohistochemistry revealed that the expression of these two genes was significantly higher in osteosarcoma than in paraneoplastic tissues. The results of qRT-PCR experiments showed that the expression of RBM15 was significantly higher in both osteosarcomas than in the control cell lines. Absolute lymphocyte value, lymphocyte percentage, hematocrit and erythrocyte count were lower in osteosarcoma than in the control group (P < 0.001). RBM15 and YTHDC1 can serve as potential prognostic biomarkers associated with m6A in osteosarcoma.© 2023. The Author(s).