Objective: To investigate the distribution and characteristics of gene mutations in osteosarcoma, and to analyze the frequency and types of detectable mutations, and to identify potential targets for individualized treatment of osteosarcoma. Methods: The fresh tissue or paraffin-embedded tissue samples of 64 cases of osteosarcoma that were surgically resected or biopsied and then subject to next generation sequencing, were collected from Beijing Jishuitan Hospital, China from November 2018 to December 2021. The tumor DNA was extracted to detect the somatic and germline mutations using targeted sequencing technology. Results: Among the 64 patients, 41 were males and 23 were females. The patient age ranged from 6 to 65 years with a median age of 17 years, including 36 children (under 18 years old) and 28 adults. There were 52 cases of conventional osteosarcoma, 3 cases of telangiectatic osteosarcoma, 7 cases of secondary osteosarcoma, and 2 cases of parosteosarcoma. The detection rate of gene mutations was overall 84.4% (54/64). There were 324 variations in 180 mutated genes, including 125 genes with copy number variations, 109 single nucleotide variants, 83 insertions or deletions, and 7 gene fusions. The most common mutated genes were TP53, VEGFA, CCND3, ATRX, MYC, RB1, PTEN, GLI1, CDK4 and PTPRD. Among them, TP53 had the highest mutation rate (21/64, 32.8%), single nucleotide variant was the main mutation type (14/23, 60.9%), and 2 cases carried the TP53 germline mutation. VEGFA and CCND3 showed copy number amplification simultaneously in 7 cases. Conclusions: The high-frequency mutation of TP53 suggests that it plays an important role in the pathogenesis and development of osteosarcoma. VEGFA, CCND3 and ATRX are mutated genes in osteosarcoma and worthy of further studies. Combination of pathologic diagnosis and next generation sequencing with clinical practice can guide individualized treatment for patients with refractory, recurrent and metastatic osteosarcoma.
目的： 探讨骨肉瘤基因突变的分布和特点，并对突变位点的频率和类型进行分析，寻找骨肉瘤个体化治疗的潜在靶点。 方法： 收集北京积水潭医院2018年11月至2021年12月接受手术切除或活检并进行二代测序检测的64例骨肉瘤的新鲜组织或石蜡包埋组织标本，提取DNA通过靶向测序技术检测肿瘤组织的体细胞突变及胚系突变基因变异情况，结合患者的临床病理特征进行分析。 结果： 64例患者中，男性41例，女性23例，发病年龄6~65岁，中位年龄17岁，其中儿童（18岁以下）36例，成人28例。骨肉瘤组织学亚型包括普通型骨肉瘤52例、毛细血管扩张型骨肉瘤3例，继发性骨肉瘤7例，骨旁骨肉瘤2例。基因突变的检出率为84.4%（54/64），共检测出180个突变基因中存在324个变异信息，包括拷贝数变异125个，点突变109个，插入或缺失83个，基因融合7个。最常见的突变基因为TP53、VEGFA、CCND3、ATRX、MYC、RB1、PTEN、GLI1、CDK4及PTPRD。其中TP53的突变比例最高（21/64，32.8%），点突变是其主要变异类型（14/23，60.9%），并有2例携带TP53胚系突变基因。VEGFA和CCND3在7例患者中同时出现拷贝数扩增。 结论： TP53的高频突变提示其在骨肉瘤的发生发展中起着重要作用，VEGFA、CCND3及ATRX等是值得进一步研究的骨肉瘤突变基因，结合临床可为难治或复发转移患者提供个体化治疗依据。.