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中国癌症防治杂志

中国癌症防治杂志 ›› 2025, Vol. 17 ›› Issue (4): 465-472.doi: 10.3969/j.issn.1674-5671.2025.04.11

• 论著 • 上一篇    下一篇

替莫唑胺获得性耐药胶质母细胞瘤细胞的脂质代谢组学分析

  

  1. 苏州大学附属第三医院核医学科； 苏州大学附属第三医院急诊外科；无锡市惠山区人民医院检验科
  • 出版日期:2025-08-25 发布日期:2025-09-12
  • 通讯作者: 贺晨 E-mail：13082510827@163.com
  • 基金资助:
    常州市科技计划项目（CJ20235086）；无锡市卫生健康委科研项目（Q202415）

Lipid metabolomics analysis of glioblastoma cells with acquired resistantance to temozolomide

  • Online:2025-08-25 Published:2025-09-12

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摘要： 目的 探讨替莫唑胺耐药性胶质母细胞瘤细胞的脂质代谢特征。 方法 选取人胶质母细胞瘤U87细胞，通过浓度递增诱导法构建对替莫唑胺耐药的细胞模型U87TR，并利用CCK⁃8法验证耐药性；分别采用平板克隆形成、Transwell侵袭和划痕愈合实验评估细胞的克隆形成、侵袭和迁移能力。采用改良版Bligh⁃Dyer法提取U87和U87TR细胞脂质，利用超高效液相色谱⁃串联质谱技术对脂质代谢物进行鉴定、定量、聚类及差异分析。采用主成分分析法和正交偏最小二乘判别（orthogonal partial least squares⁃discriminant analysis，OPLS⁃DA）法筛选与替莫唑胺获得性耐药相关的脂质代谢物。 结果 成功构建了替莫唑胺获得性耐药U87TR细胞模型。与敏感亲本细胞相比，U87TR细胞表现出显著增强的克隆形成、侵袭、迁移能力（均P<0.05）。脂质代谢组学分析显示，两种细胞共鉴定出32亚类(主要包括磷脂酰胆碱、甘油三酯和鞘磷脂等)共417种脂质代谢物，其中260个脂质代谢物含量的差异具有统计学意义。与U87细胞相比，U87TR细胞中PG34∶2(18∶1_16∶1)、PG38∶5(18∶1_20∶4)、BMP34∶3(18∶2_16∶1)的表达水平显著降低（均P<0.05）。相反，PG34∶2(18∶2_16∶0)、PG36∶4(18∶2_18∶2)、PG36∶3(18∶2_18∶1)等的表达水平则显著升高（均P<0.05）。OPLS⁃DA进一步证实，PI36∶3(18∶2_18∶1)、PA32∶0、LacCer d18∶1/14∶0等脂质代谢物能有效区分耐药细胞和敏感细胞。 结论 替莫唑胺获得性耐药胶质母细胞瘤细胞相较于敏感细胞表现出独特的脂质代谢特征，这可能与在胶质母细胞瘤中观察到的药物耐受性相关。

关键词: 胶质母细胞瘤, 脂质组学, 替莫唑胺, 耐药, 超高效液相色谱-串联质谱

Abstract: Objective To investigate the alterations in lipid metabolism characteristics of temozolomide⁃resistant glioblastoma cells. Methods Human glioblastoma U87 cells were utilized to develop a temozolomide⁃resistant cell model (U87TR) through a concentration gradient induction method, with resistance confirmed by CCK⁃8 assay. The clonogenic, invasive, and migration abilities of the cells were assessed using the plate colony formation assay, Transwell invasion assay, and wound healing assay, respectively. Lipids were extracted from both U87 and U87TR cells employing a modified Bligh⁃Dyer method, and lipid metabolites were subsequently identified, quantified, clustered, and analyzed for differences using ultra high performance liquid chromatography⁃tandem mass spectrometry. Principal component analysis and orthogonal partial least squares⁃discriminant analysis (OPLS⁃DA) were utilized to identify lipid metabolites associated with temozolomide⁃acquired resistance. Results The temozolomide⁃acquired resistant U87TR model was successfully established. Compared to the sensitive parental cells, U87TR cells demonstrated significantly enhanced clonogenic formation, invasion, and migration capabilities (all P<0.05). Lipidomic profiling identified a total of 417 lipid metabolites in 32 categories (mainly including phosphatidylcholines, triglycerides, and sphingomyelin), among which 260 lipid metabolites showed statistically significant differences in content. The expression levels of PG34∶2(18∶1_16∶1), PG38∶5 (18∶1_20∶4), and BMP34∶3 (18∶2_16∶1) were significantly decreased in U87TR cells compared to U87 cells (all P<0.05). Conversely, the expression levels of PG34∶2 (18∶2_16∶0), PG36∶4 (18∶2_18∶2), PG36∶3 (18∶2_18∶1)， etc. were significantly increased (all P<0.05). OPLS⁃DA confirmed that lipid metabolites such as PI36∶3 (18∶2_18∶1), PA32∶0, and LacCer d18∶1/14∶0 effectively differentiate resistant cells from sensitive ones. Conclusions Temozolomide⁃resistant glioblastoma cells exhibit distinct lipid metabolite profiles compared to sensitive cells，which may be associated with the drug resistance observed in  glioblastoma.

Key words: Glioblastoma, Lipidomics, Temozolomide, Drug resistance； Ultra high performance liquid chromatography?tandem mass spectrometry 

中图分类号: 

  • R739.41