TP53 Sanger sequencing for osteosarcoma in .ab1 files

Osteosarcoma (OS) is the most common malignant bone tumor affecting adolescents and young adults. The detection of cancer-related genetic alterations has a growing impact to guide diagnosis, prognosis, and targeted therapies. In this study, Sanger sequencing was performed to investigate somatic and germline <i>TP53</i> mutations (exons 4-8) in an adolescent patient with OS. No PCR products for <i>TP53</i> exon 5 were detected in the tumor sample by PCR analysis prior to Sanger sequencing, suggesting a significant deletion in this exon. Sanger sequencing analysis revealed the missense variant <i>TP53</i> c.712T&gt;A (p.Cys238Ser) in tumor tissue sample and the <i>TP53 </i>c.215C&gt;G (p.Pro72Arg) germline missense variant was identified in the peripheral blood sample. Taken together, these findings reinforce the idea that <i>TP53</i> mutations represent key oncogenic drivers in OS patients.

骨肉瘤（Osteosarcoma, OS）是最常见的累及青少年与年轻成人的恶性骨肿瘤。癌症相关遗传变异的检测对指导诊断、预后评估及靶向治疗的影响日益加深。本研究采用桑格测序（Sanger sequencing）技术，对1例骨肉瘤青少年患者的体细胞及生殖系TP53基因外显子4-8开展突变检测。在桑格测序前的聚合酶链式反应（Polymerase Chain Reaction, PCR）分析中，未在肿瘤样本中检测到TP53基因外显子5的PCR产物，提示该外显子存在显著缺失。桑格测序分析显示，肿瘤组织样本中存在TP53基因c.712T>A（p.Cys238Ser）错义变异，外周血样本中检出TP53基因c.215C>G（p.Pro72Arg）生殖系错义变异。综上，上述研究结果进一步证实，TP53突变是骨肉瘤患者关键的致癌驱动因素。