目的:髓母细胞瘤(MB),儿科患者常见且异质的后窝肿瘤,呈现不同的预后结果。为了提高我们对MB复杂生物学的理解,具有必要数据的新型患者肿瘤来源培养物MB模型的开发仍然是一项基本要求.
方法:我们在体外连续传代PUMC-MB1,以建立连续的细胞系。我们使用细胞计数试剂盒-8(CCK-8)检查了体外生长,并使用皮下和颅内异种移植模型检查了体内生长。用苏木精和伊红(HE)染色和免疫组织化学(IHC)对异种移植物进行组织病理学研究。同时,我们使用全基因组测序(WGS)探索了它的分子特征,靶向测序,和RNA测序。在生物信息学分析的指导下,我们在体外和体内验证了PUMC-MB1的药物敏感性。
结果:PUMC-MB1,来自高危MB患者,显示种群倍增时间(PDT)为48.18h,并在20天内在SCID小鼠中实现了100%的肿瘤生长。原始肿瘤和异种移植物的HE和免疫组织化学检查证实了PUMC-MB1作为经典MB的分类。通过WGS进行的基因组分析显示MYC和OTX2扩增同时发生。RNA-seq数据将其分类为第3组MB亚组,而根据世界卫生组织的分类,它属于非WNT/非SHHMB。与D283和D341med的比较分析确定了4065个差异表达基因,在PI3K-AKT途径中具有显著的富集。顺铂,4-氢过氧环磷酰胺/环磷酰胺,长春新碱,和dactolisib(选择性PI3K/mTOR双重抑制剂)在体外和体内显着抑制PUMC-MB1的增殖。
结论:PUMC-MB1,一种新的第3组(非WNT/非SHH)MB细胞系,具有全面的增长特征,病理学,和分子特征。值得注意的是,dactolisib表现出有效的抗增殖作用,毒性最小,承诺一个潜在的治疗途径。PUMC-MB1可以作为揭示MB机制和创新治疗策略的有价值的工具。
OBJECTIVE: Medulloblastoma (MB), a common and heterogeneous posterior fossa tumor in pediatric patients, presents diverse prognostic outcomes. To advance our understanding of MB\'s intricate biology, the development of novel patient tumor-derived culture MB models with necessary data is still an essential requirement.
METHODS: We continuously passaged PUMC-MB1 in vitro in order to establish a continuous cell line. We examined the in vitro growth using Cell Counting Kit-8 (CCK-8) and in vivo growth with subcutaneous and intracranial xenograft models. The xenografts were investigated histopathologically with Hematoxylin and Eosin (HE) staining and immunohistochemistry (IHC). Concurrently, we explored its molecular features using Whole Genome Sequencing (WGS), targeted sequencing, and RNA sequecing. Guided by bioinformatics analysis, we validated PUMC-MB1\'s drug sensitivity in vitro and in vivo.
RESULTS: PUMC-MB1, derived from a high-risk MB patient, displayed a population doubling time (PDT) of 48.18 h and achieved 100% tumor growth in SCID mice within 20 days. HE and Immunohistochemical examination of the original tumor and xenografts confirmed the classification of PUMC-MB1 as a classic MB. Genomic analysis via WGS revealed concurrent MYC and OTX2 amplifications. The RNA-seq data classified it within the Group 3 MB subgroup, while according to the WHO classification, it fell under the Non-WNT/Non-SHH MB. Comparative analysis with D283 and D341med identified 4065 differentially expressed genes, with notable enrichment in the PI3K-AKT pathway. Cisplatin, 4-hydroperoxy cyclophosphamide/cyclophosphamide, vincristine, and dactolisib (a selective PI3K/mTOR dual inhibitor) significantly inhibited PUMC-MB1 proliferation in vitro and in vivo.
CONCLUSIONS: PUMC-MB1, a novel Group 3 (Non-WNT/Non-SHH) MB cell line, is comprehensively characterized for its growth, pathology, and molecular characteristics. Notably, dactolisib demonstrated potent anti-proliferative effects with minimal toxicity, promising a potential therapeutic avenue. PUMC-MB1 could serve as a valuable tool for unraveling MB mechanisms and innovative treatment strategies.