Background: MicroRNAs (miRNAs) function as post-transcriptional gene expression regulators and influence the development and progression of several cancers, yet their roles in pediatric sarcomas remain poorly defined. Methods: RNA extracted from formalin-fixed paraffin-embedded tumor tissue scrolls of 108 pediatric tumors, including 32 osteosarcoma (OS), 26 Ewing’s sarcoma (EWS), and 50 rhabdomyosarcoma (RMS) cases, were analyzed for microRNA expression using the NanoString multiplex nCounter platform that yielded information on 827 human miRNAs. The expression of candidate miRNAs was validated with in situ hybridization (miRNA-ISH) and QuPath quantification on tissue microarray slides comprising an independent set of 48 OS, 17 EWS, and 104 RMS adult and pediatric cases collectively. Results: The differential expression analysis of nCounter data identified 23 miRNAs enriched in RMS, 33 in EWS, and 45 in OS (>3 fold change andp< 0.01). miR-206 was most strongly associated (>55 fold change,p< 1 × 10−9) with RMS and demonstrated the highest sensitivity and specificity in distinguishing RMS from EWS and OS; this finding was also confirmed by miRNA-ISH. A combined signature of differentially expressed miRNAs reliably separated alveolar from embryonal RMS. The expression of miR-9-5p in EWS and miR-140-5p in OS discriminated among the different tumors and correlated with adverse patient outcome. The nCounter assay exhibited greater sensitivity than miRNA-ISH in detecting miR-206 and miR-140-5p expression. Conclusions: Collectively, these findings demonstrate that distinct miRNA profiles can differentiate pediatric sarcoma types and subtypes and offer clinically relevant insights into tumor biology, prognosis, and potential diagnostic application.
背景：微小RNA（miRNA）作为转录后基因表达调控因子，在多种癌症的发生发展中发挥作用，但其在儿童肉瘤中的具体功能尚不明确。方法：本研究通过NanoString多重nCounter平台对108例儿童肿瘤（包括32例骨肉瘤、26例尤文肉瘤和50例横纹肌肉瘤）的石蜡包埋组织切片进行RNA提取，检测了827种人类miRNA的表达谱。采用原位杂交技术和QuPath定量分析，在包含48例骨肉瘤、17例尤文肉瘤及104例横纹肌肉瘤（涵盖成人与儿童病例）的组织芯片上对候选miRNA进行独立验证。结果：nCounter数据的差异表达分析显示，横纹肌肉瘤中富集23种miRNA，尤文肉瘤中富集33种，骨肉瘤中富集45种（变化倍数>3且p<0.01）。其中miR-206与横纹肌肉瘤的关联性最强（变化倍数>55，p<1×10−9），在区分横纹肌肉瘤与尤文肉瘤、骨肉瘤时表现出最高的敏感性和特异性，该结果亦通过原位杂交得到证实。差异表达miRNA组合特征可有效区分腺泡状与胚胎性横纹肌肉瘤亚型。尤文肉瘤中的miR-9-5p和骨肉瘤中的miR-140-5p表达水平具有肿瘤特异性，且与患者不良预后相关。在检测miR-206和miR-140-5p表达时，nCounter技术较原位杂交显示出更高灵敏度。结论：本研究表明，特征性miRNA谱可有效鉴别儿童肉瘤类型及亚型，为肿瘤生物学机制、预后评估及潜在诊断应用提供了具有临床价值的见解。
MicroRNA Profiling Identifies Diagnostic and Prognostic Markers in Pediatric Sarcoma
肿瘤(癌症)患者之家