The engagement of Ras/Raf/MEK/ERK and PLCγ1/PKC pathways regulated by TrkB receptor in resistance of glioma cells to elimination upon apoptosis induction

IF 4.6 2区医学 Q1 NEUROSCIENCES

Neuropharmacology Pub Date : 2024-11-07 DOI:10.1016/j.neuropharm.2024.110204

Adrian Zając , Joanna Sumorek-Wiadro , Aleksandra Maciejczyk , Michał Chojnacki , Iwona Wertel , Wojciech Rzeski , Joanna Jakubowicz-Gil

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Abstract

The most aggressive tumors of human central nervous system are anaplastic astrocytoma (AA, III grade) and glioblastoma multiforme (GBM, IV grade) with an extremely bad prognosis. Their malignant character and resistance to standard therapy are correlated to the over-expression of survival pathways such as Ras/Raf/MEK/ERK and PLCγ1/PKC regulated by TrkB receptor. Therefore, the aim of this study was to investigate the engagement of those pathways in human glioma cells resistance for apoptosis induction by Temozolomide treatment. Two cancer MOGGCCM (AA) and T98G (GBM) and normal human astrocytes (NHA) cell lines were utilized. The tested inhibitors single and simultaneous action with Temozolomide affection on apoptosis induction was analyzed by MTT, microscopic observations and flow cytometry. Bcl-2:beclin-1 complexes occurrence was also assessed. siRNAs were used for direct proof of tested pathways engagement in gliomas resistance to apoptosis elimination. The most effective in eliminating gliomas with minimal astrocyte damage was 5 μM PLCγ1 inhibitor (U-73122) for MOGGCCM and 15 μM for T98G cells, and 1 μM LOXO-101 for all cancer cells. Sorafenib, Temozolomide, U-73122, and LOXO-101 effectively eliminate cancer cells. Single applications of sorafenib and Temozolomide were effective, but had lower efficiency than U-73122 and LOXO-101. These drugs induced apoptosis, affecting mitochondrial membrane potential and caspases 3, 8, and 9 activity. The study found that a Bcl-2:beclin-1 complex formation was observed when apoptosis was dominant. Inhibiting the pathways regulated by TrkB receptor combined with Temozolomide action, led to successful gliomas elimination. Those results might serve as basis for modern targeted treatment development.

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受 TrkB 受体调控的 Ras/Raf/MEK/ERK 和 PLCγ1/PKC 通路参与了胶质瘤细胞凋亡诱导过程中的抗消除过程。

人类中枢神经系统最具侵袭性的肿瘤是预后极差的无弹性星形细胞瘤（AA，III 级）和多形性胶质母细胞瘤（GBM，IV 级）。它们的恶性特征和对标准疗法的抗药性与受 TrkB 受体调控的 Ras/Raf/MEK/ERK 和 PLCγ1/PKC 等生存通路的过度表达有关。因此，本研究旨在探讨这些通路在人类胶质瘤细胞抗替莫唑胺治疗诱导凋亡过程中的参与情况。本研究利用了两种癌症细胞株 MOGGCCM（AA）和 T98G（GBM）以及正常人星形胶质细胞（NHA）。通过 MTT、显微镜观察和流式细胞术分析了所测试的抑制剂对诱导细胞凋亡的单一或与替莫唑胺同时作用的影响。还评估了Bcl-2:beclin-1复合物的发生情况。使用siRNA直接证明了所测试的通路参与胶质瘤对凋亡消除的抵抗。对 MOGGCCM 和 T98G 细胞分别使用 5 μM 和 15 μM 的 PLCγ1 抑制剂（U-73122）以及 1 μM 的 LOXO-101 对所有癌细胞最有效。索拉非尼、替莫唑胺、U-73122 和 LOXO-101 能有效消除癌细胞。单次使用索拉非尼和替莫唑胺也有效，但效率低于 U-73122 和 LOXO-101。这些药物可诱导细胞凋亡，影响线粒体膜电位和 Caspases 3、8 和 9 的活性。研究发现，当细胞凋亡占主导地位时，会观察到 Bcl-2：beclin-1 复合物的形成。抑制 TrkB 受体调控的途径与替莫唑胺的作用相结合，成功地消除了胶质瘤。这些结果可作为现代靶向治疗开发的基础。

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来源期刊

Neuropharmacology 医学-神经科学

CiteScore

10.00

自引率

4.30%

发文量

288

审稿时长

45 days

期刊介绍： Neuropharmacology publishes high quality, original research and review articles within the discipline of neuroscience, especially articles with a neuropharmacological component. However, papers within any area of neuroscience will be considered. The journal does not usually accept clinical research, although preclinical neuropharmacological studies in humans may be considered. The journal only considers submissions in which the chemical structures and compositions of experimental agents are readily available in the literature or disclosed by the authors in the submitted manuscript. Only in exceptional circumstances will natural products be considered, and then only if the preparation is well defined by scientific means. Neuropharmacology publishes articles of any length (original research and reviews).