通过点印迹筛选确定替莫唑胺调控蛋白作为胶质瘤多指标组学研究的潜在靶点

IF 3.3 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Frontiers in bioscience (Landmark edition) Pub Date : 2024-10-22 DOI:10.31083/j.fbl2910364
Anna Maria Bielecka-Wajdman, Grzegorz Machnik, Christina Linnebacher, Michael Linnebacher, Katarzyna Stec-Grosman, Ewa Obuchowicz
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引用次数: 0

摘要

背景:恶性胶质瘤是脑癌中的一种异质性肿瘤,其特点是浸润性生长,肿瘤边界不清晰可辨。由于无法进行根治性手术切除和靶向治疗,因此预后较差。尽管替莫唑胺(TMZ)仍是治疗胶质瘤的主要化疗药物,但由于肿瘤耐药性的产生,其疗效有限。因此,改善这些肿瘤的诊断和治疗刻不容缓。寻找和开发胶质瘤的特异性生物标志物对确定治疗目标、监测治疗以及构建个性化疗法都很有用。然而,目前仍没有可靠的标志物能改变胶质瘤治疗的质量:本研究采用点印迹法分析了三种胶质瘤细胞系中 84 种癌症相关蛋白的表达差异:商用 T98G 细胞和两种患者衍生细胞系。研究还调查了 TMZ 对蛋白质表达、细胞形态和迁移变化的影响(Proteome Profiler Human XL Oncology Array、LeviCell System、显微成像)。被分析的细胞系在蛋白质表达和蛋白质组变化方面具有显著不同的可塑性,而这些变化是由 TMZ 诱导的:点印迹分析显示,所有细胞系共有十种蛋白质,其中五种(Cathepsin b、FGF、Survivin、AXL、Osteopontin)受 TMZ 影响。由于暴露于 TMZ,在 HROG02 和 T98G 细胞系中都检测到了参与化疗抗性和侵袭的蛋白质(TIE-2、Thrombospondin)。在 HROG17 细胞的对照培养物(未暴露于 TMZ)中,参与新陈代谢的蛋白质受到强烈抑制:本文提供的数据揭示了替莫唑胺对一组蛋白质表达的调节作用:结论:本研究数据揭示了替莫唑胺对一组蛋白质表达的调节作用,这些蛋白质包括:Cathepsin b、成纤维细胞生长因子(FGF)、Survivin、AXL和Osteopontin。不过,还需要对体液中的蛋白质进行进一步的高通量分析和检测。
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A Dot-Blot Screening for Identifying the Temozolomide-Regulated Proteins as Potential Targets for Glioma Multi-OMICS Studies.

Background: Malignant gliomas represent a heterogenous group of brain cancers that are characterized by infiltrative growth that lacks a clearly identifiable tumor border. The lack of the possibility of radical surgical resection and targeted therapy results in a poor prognosis. Although Temozolomide (TMZ) is still the leading chemotherapeutic agent in glioma treatment, its efficacy is limited due to the development of tumor resistance. Therefore, there an urgent need to improve the diagnosis and treatment of these tumors. Finding and developing biomarkers that are specific to glioma could be useful for both identifying therapy targets and monitoring treatment as well as for constructing a personalized therapy. However, there are still no reliable markers that would change the quality of glioma treatment.

Methods: In this study, differences in the expression of 84 cancer-related proteins in three glioma cell lines were analyzed using the dot-blot method: commercial T98G cells and two patient-derived cell lines. The influence of TMZ on changes in protein expression, cell morphology, and migration was also investigated (Proteome Profiler Human XL Oncology Array, LeviCell System, Microscopic imaging). The lines that were analyzed were characterized by a remarkably different plasticity of protein expression and the proteomic alterations that were induced by TMZ.

Results: A dot-blot analysis revealed ten proteins that were common to all of the lines and five (Cathepsin b, FGF, Survivin, AXL, Osteopontin) that were modulated by the TMZ. As a result of the exposure of TMZ, the proteins that are involved in chemoresistance and invasion (TIE-2, Thrombospondin) were detected in both the HROG02 and T98G cell lines. In the control culture (not exposed to TMZ) of HROG17 cells, the proteins that are involved in metabolism were strongly suppressed.

Conclusions: The presented data sheds new light on the modulatory effect of Temozolomide on the expression of a protein panel: Cathepsin b, fibroblast growth factor (FGF), Survivin, AXL, and Osteopontin that may suggest their potential as therapeutic targets or biomarkers to monitoring therapy effects. However, further high-throughput analysis and detection of the proteins in the body fluids are necessary.

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