首都医科大学学报 ›› 2023, Vol. 44 ›› Issue (5): 733-740.doi: 10.3969/j.issn.1006-7795.2023.05.005

• 恶性肿瘤诊治进展 • 上一篇    下一篇

人胶质母细胞瘤细胞系Y1203的构建及应用

郑义1,宋莱荣2,徐晓颖2,吴震2,李晓杰2,解磊3,陈峰1,李文斌1*   

  1. 1.首都医科大学附属北京天坛医院肿瘤综合治疗中心, 北京 100070; 2.首都医科大学附属北京天坛医院神经外科,北京 100070;3.河北医科大学附属第三医院神经外科,石家庄 050051
  • 收稿日期:2023-07-18 出版日期:2023-10-20 发布日期:2023-10-26
  • 通讯作者: 李文斌 E-mail:liwenbin@ccmu.edu.cn
  • 基金资助:
    国家自然科学基金项目(81972338),首都医科大学附属北京天坛医院人才引进项目(RCYJ-2020-2025-LWB),首都医科大学附属北京天坛医院横向课题

Construction and application of the human glioblastoma cell line Y1203

Zheng Yi1, Song Lairong2, Xu Xiaoying2, Wu Zhen2, Li Xiaojie2, Xie Lei3, Chen Feng1, Li Wenbin1*   

  1. 1.Department of Neuro-Oncology, Cancer Center, Beijing Tiantan Hospital, Capital Medical University, Beijing 100070, China;2. Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, Beijing 100070, China;3.Department of Neurosurgery, The Third Affiliated Hospital of Hebei Medical University, Shijiazhuang 050051, China
  • Received:2023-07-18 Online:2023-10-20 Published:2023-10-26
  • Supported by:
    this study was supported by National Natural Science Foundation of China(81972338),Beijing Tiantan Hospital, Capital Medical University Talent Recruitment(RCYJ-2020-2025-LWB), Horizontal Scientific Research in Beijing Tiantan Hospital,Capital Medical University.

摘要: 目的  构建一株具备干细胞表型的胶质母细胞瘤细胞系Y1203,用于研究肿瘤细胞干性维持以及治疗耐药的潜在机制。方法  Y1203细胞来源于一位49岁女性胶质母细胞瘤患者复发再手术切除的肿瘤组织。使用Y1203细胞构建人源胶质母细胞瘤原位及皮下移植(patient derived tumor xenograft, PDX)模型。采用流式细胞术检测不同代数Y1203细胞的细胞周期。采用实时荧光定量聚合酶链反应检测Y1203细胞的干细胞标志物转录因子SOX-2(transcription factor SOX-2, SOX2)和细胞黏附分子分化抗原簇-44(cluster of differentiation-44, CD44)表达。基于转录组测序技术(RNA sequencing,RNA-Seq)比较Y1203细胞和U87细胞表达谱差异。结果  短串联重复序列(short tandem repeats,STR)鉴定确认Y1203细胞与患者肿瘤组织同源,且其与ExPASy数据库中涵盖的常见细胞系完全不同。基因检测结果提示异柠檬酸脱氢酶1(isocitrate dehydrogenase 1,IDH1)无突变,端粒酶反转录酶(telomerase reverse transcriptase,TERT)启动子突变,原癌基因(B-Raf proto-oncogene,BRAF V600E)、抑癌基因(tumor protein p53,TP53)和转录调控因子(ATRX chromatin remodeler,ATRX)基因突变。人胶质母细胞瘤细胞系Y1203可实现在体外无限增殖并用于建立PDX模型。与其他胶质瘤细胞系相比,Y1203细胞中高表达的肿瘤干细胞标志物为SOX2和CD44,并显著富集胶质瘤干细胞相关通路。结论  人胶质母细胞瘤细胞系Y1203分化低,恶性程度较高,对放射性治疗和化学治疗抵抗性强,具有胶质母细胞瘤干细胞特性。该细胞系的建立为胶质母细胞瘤临床前研究提供了实验基础。

关键词: 胶质母细胞瘤, 原代培养, 干细胞特性, 胶质母细胞瘤原位及皮下移植模型

Abstract: Objective  To establish a glioblastoma cell line, Y1203, with stem cell-like phenotypes for the study of tumor cell stemness maintenance and potential mechanisms of treatment resistance. Methods  Y1203 cells were derived from tumor tissue obtained by recurrent resection in a 49-year-old female glioblastoma patient. Y1203 cells were used to establish patient-derived tumor xenograft (PDX) models both orthotopically and subcutaneously. Flow cytometry was applied to analyze the cell cycle distribution of different passages of Y1203 cells. Real-time quantitative polymerase chain reaction was performed to assess the expression of stem cell markers, including transcription factor SOX-2 (SOX2) and cluster of differentiation-44 (CD44). RNA-Seq was conducted to compare the expression profiles between Y1203 and U87 cells. Results  The short tandem repeats (STR) analysis confirmed the homogeneity between Y1203 cells and the patient’s tumor tissue, and they were distinct from commonly used cell lines available in the ExPASy database. Genetic testing revealed isocitrate dehydrogenase 1 (IDH1) wild-type, telomerase reverse transcriptase (TERT) promoter mutation, B-Raf proto-oncogene (BRAF V600E) mutation, tumor protein p53 (TP53) mutation, and ATRX chromatin remodeler (ATRX) mutation. The human glioblastoma cell line Y1203 exhibited unlimited proliferation in vitro and thus orthotopic and subcutaneous glioblastoma models was successfully established. Compared to other glioma cell lines, Y1203 cells showed high expression of the tumor stem cell markers CD44 and SOX2, as well as significant enrichment of glioma stem cell-associated pathways. Conclusions  The human glioblastoma cell line Y1203 shows low differentiation, high malignancy, and strong resistance to radiotherapy and chemotherapy, and it possesses glioblastoma stem cell characteristics. The establishment of Y1203 cell line provides an experimental basis for preclinical studies of glioblastoma.

Key words: glioblastoma, primary culture, stemness, patient derived tumor xenograft model

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