Background/Objectives: Osteosarcoma is an aggressive bone malignancy with high metastatic potential to the lungs. CTCs, as seeds of metastasis, play an important role in the spread of this cancer, and, therefore, their isolation, culture, and gene expression analysis promises valuable insights into the progression and metastatic cascade of osteosarcoma. The aim of this study was to isolate and culture CTCs from osteosarcoma-bearing mice and compare their migration, radioresistance, and gene expression with their parental cell line. Methods: CTCs from LM8-inoculated mice were isolated and cultured. The gene expression of the CTC-derived cell lines was then compared to the parental cell line. Furthermore, a Transwell assay, a clonogenic assay after irradiation, and immunohistochemical stainings were used to compare the CTC-derived cell lines with the parental cell line. Results: The CTC-derived cell lines differed significantly in gene expression from their parental cell line. 361 differentially expressed genes were identified, among which GO and KEGG analysis revealed major differences in the expression of genes related to antigen processing and presentation and extracellular matrix constituents. In addition, the CTC-derived cell lines were observed to have a higher migratory capacity and comparable radioresistance compared to the parental cell line. CD44 expression was found to be conserved in CTC-derived cell lines. Conclusions: This study provides a comparison between CTC-derived and their parental cell lines in terms of gene expression, migration, and radioresistance. Our findings allow for further research in the field of osteosarcoma CTCs and their generation. Furthermore, the identified DEGs between CTCs and their parental cell line can serve as a reference point for targeted therapies against osteosarcoma CTCs.
背景/目的：骨肉瘤是一种具有高肺转移倾向的侵袭性骨恶性肿瘤。循环肿瘤细胞作为转移的种子，在该癌症的扩散中起着重要作用，因此，对其分离、培养和基因表达分析有望为骨肉瘤的进展和转移级联提供有价值的见解。本研究旨在从荷骨肉瘤小鼠中分离和培养循环肿瘤细胞，并将其迁移能力、放射抗性和基因表达与亲本细胞系进行比较。方法：分离并培养接种LM8小鼠的循环肿瘤细胞。随后将循环肿瘤细胞衍生细胞系的基因表达与亲本细胞系进行比较。此外，采用Transwell实验、照射后克隆形成实验以及免疫组织化学染色法，比较循环肿瘤细胞衍生细胞系与亲本细胞系。结果：循环肿瘤细胞衍生细胞系的基因表达与亲本细胞系存在显著差异。共鉴定出361个差异表达基因，其中GO和KEGG分析显示，与抗原加工呈递和细胞外基质成分相关的基因表达存在主要差异。此外，与亲本细胞系相比，循环肿瘤细胞衍生细胞系表现出更高的迁移能力和相当的放射抗性。研究发现CD44表达在循环肿瘤细胞衍生细胞系中得以保留。结论：本研究在基因表达、迁移能力和放射抗性方面对循环肿瘤细胞衍生细胞系及其亲本细胞系进行了比较。我们的发现为骨肉瘤循环肿瘤细胞及其生成领域的进一步研究提供了基础。此外，所鉴定的循环肿瘤细胞与亲本细胞系之间的差异表达基因可作为针对骨肉瘤循环肿瘤细胞靶向治疗的参考依据。
肿瘤(癌症)患者之家