摘要/Abstract

摘要： 目的 ·建立 1株新的中国汉族人视网膜母细胞瘤细胞系，并研究其基本特征。方法 ·在无菌环境中，从 1例视网膜母细胞瘤患者瘤体标本中分离原代肿瘤细胞，进行培养、纯化与传代。通过细胞免疫荧光、免疫组织化学染色、 DNA短片段重复序列 (short tandem repeat，STR)检测、核型分析和全外显子测序等方法，观察并鉴定该细胞系和肿瘤组织形态学特征、遗传学特征及表面标志物。结果 ·将该细胞系命名为 SNPH-Rb-C24，其主要特征为神经起源的肿瘤细胞，表达神经细胞黏附分子 1和突触素； STR分型与原肿瘤组织个体来源一致；染色体核型发生复杂改变，但未发现 13号染色体长臂改变；与原肿瘤组织皆表现为 RB1+/+特征，且基因突变谱相似；具有体内成瘤能力。结论 ·成功建立 1株起源于 RB1+/+中国汉族人视网膜母细胞瘤细胞系 SNPH-Rb-C24，该细胞系保留了原肿瘤组织的生物学和遗传学特征。
关键词: 视网膜母细胞瘤, 原代细胞系, RB1基因, 核型分析, 全外显子测序
Abstract:
Objective · To develop and characterize a novel retinoblastoma cell line a Han Chinese patient. Methods · Cells were dispersed tumor tissue harvested an enucleated eyeball harbouring retinoblastoma under sterile conditions. The dissociated cells were cultured, purified and passaged in vitro. Morphologic and genetic analysis and detection of surface biomarkers were performed on the cell line and primary tumorimmunofluorescence, immunohistochemical staining, DNAshort tandemrepeat (STR) analysis, karyotype analysis, and exome sequencing. Results · This human retinoblastoma cell line was designated as SNPH-Rb-C24. It expressed neural cell adhesion molecule 1 and synaptophysin, which confirmed its neuronal derivation. DNA STR analysis showed an identical match between SNPH-Rb-C24 and primary tumor. Karyotype analysis showed complex chromosomal abnormalities in SNPH-Rb-C24, while no alteration in 13q was observed. Comparative exome sequencing identified common mutated genes and RB1+/+ in both SNPH-Rb-C24 and primary tumor. Orthotopic xenograft tumors derived early passage cells were established. Conclusion · A human retinoblastoma cell line (SNPH-Rb-C24) derived a Han Chinese patient with RB1+/+ retinoblastoma is developed, which retains critical biological and genomic features of the donor tumor.
Key words: retinoblastoma, primary cell line, RB1 gene, karyotypeanalysis, exome sequencing


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