PO.MCB10.01 · 分子与细胞生物学
跨物种microRNA表达模式鉴定骨肉瘤中的潜在治疗靶点
Cross-species microRNA expression patterns identify potential therapeutic targets in osteosarcoma
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
骨肉瘤是人类和犬类中最常见的原发性骨肿瘤。在两个物种中，骨肉瘤均显示出明显的肺转移倾向，使犬类成为该病稳健且具有临床相关性的转化模型。在当前标准治疗下，局限性骨肉瘤患者的5年生存率约为60-70%，但对于就诊时即为转移性疾病的患者则降至约20-30%。在犬类中，截肢后化疗的中位生存时间约为10-12个月，不足20%的患者存活超过两年。尽管支持治疗有渐进改善，但在两个物种中，骨肉瘤的治疗策略在三十多年里基本未变，凸显了对新型预后指标和治疗靶点的需求。microRNA（miRNA）是小的非编码RNA，可作为抑癌基因或癌基因发挥作用。它们在多种组织中表达，且已知在癌症中出现失调。在本研究中，评估了20个骨肉瘤细胞系（6个人类和14个犬类），源自原发和转移性肿瘤，通过一系列体外功能实验评估关键恶性行为。这些实验包括增殖（倍增时间）、迁移（划痕和transwell实验）、非贴壁存活（失巢凋亡实验）、侵袭（滴落侵袭实验）和集落形成（克隆形成实验）的测量。随后从每个细胞系中分离RNA，并使用实时定量聚合酶链反应（RT-qPCR）定量48个候选miRNA的表达。使用Pearson's r和错误发现率（FDR）分析，将每个miRNA与所测行为进行相关分析。经FDR校正后，10个miRNA在p<0.05水平上与至少一种行为显著相关。值得注意的是，miR-708-5p与多种恶性表型显著相关，包括倍增时间、迁移和集落形成。因此，miRNA表达谱揭示了几个与恶性行为相关的候选miRNA，提示部分miRNA可能促进疾病进展，并代表未来治疗开发的有前景靶点。
查看英文原文 English abstract
Osteosarcoma is the most common primary bone tumour in both humans and dogs. In both species, osteosarcoma displays a marked propensity for pulmonary metastasis, making canines a robust and clinically relevant translational model of the disease. With current standard-of-care treatment, the 5-year survival rate for humans with localized osteosarcoma is approximately 60-70%, but falls to around 20-30% for patients presenting with metastatic disease. In dogs, amputation followed by chemotherapy yields a median survival time of roughly 10-12 months, with fewer than 20% surviving beyond two years. Despite incremental improvements in supportive care, therapeutic strategies for osteosarcoma have remained largely unchanged for more than three decades in both species, highlighting the need for novel prognostic indicators and therapeutic targets. MicroRNAs (miRNAs) are small, non-coding RNAs that can function as tumour suppressors or oncogenes. They are expressed across diverse tissues and are known to exhibit dysregulation in cancer. In this study, 20 osteosarcoma cell lines (6 human and 14 canine), derived from both primary and metastatic tumours, were evaluated for key malignant behaviours using a series of in vitro functional assays. These included measurements of proliferation (doubling time), migration (scratch and transwell assays), anchorage-independent survival (anoikis assay), invasion (drop invasion assay), and colony formation (clonogenic assay). RNA was subsequently isolated from each cell line, and expression of 48 candidate miRNAs was quantified using real-time quantitative polymerase chain reaction (RT-qPCR). Each miRNA was correlated with measured behaviours using Pearson's r and false discovery rate (FDR) analysis. Ten miRNAs were significantly associated with at least one behaviour at p < 0.05 following FDR correction. Notably, miR-708-5p demonstrated significant correlations with multiple malignant phenotypes, including doubling time, migration, and colony formation. MiRNA expression profiles, therefore, revealed several candidates associated with malignant behaviour, suggesting that select miRNAs may contribute to disease progression and represent promising targets for future therapeutic development.
利益披露 Disclosure
S. C. Fernandes, None..
E. N. Vanderboon, None..
A. Viloria-Petit, None..
R. Wood, None..
G. A. Wood, None.