Molecular Signaling Lab, Faculty of Medicine and Health Technology, Tampere University, Tampere 33720, Finland; BioMeditech and Tays Cancer Center, Tampere University Hospital, P.O. Box 553, 33101 Tampere, Finland.
Molecular Signaling Lab, Faculty of Medicine and Health Technology, Tampere University, Tampere 33720, Finland; Department of Biotechnology, Lady Doak College, Thallakulam, Madurai 625002, India.
Life Sci. 2022 Feb 15;291:120307. doi: 10.1016/j.lfs.2022.120307. Epub 2022 Jan 10.
Glioblastoma multiforme (GBM) is the most common and aggressive primary adult brain tumor. GBM is characterized by a heterogeneous population of cells that are resistant to chemotherapy. Recently, we have synthesized CHBC, a novel indole derivative targeted to GBM biomarker G-protein-coupled receptor 17 and inhibitor of GBM cells. In this study, CHBC was further investigated to characterize the efficiency of this agonist at the molecular level and its underlying mechanism in GBM cell death induction.
The effect of CHBC and TMZ was determined using time dependent inhibitor assay in glioblastoma cells, LN229 and SNB19. Drug induced cell cycle arrest was measured using PI staining followed by image analysis. The induction of apoptosis and mechanism of action of CHBC was studied using apoptosis, caspase 3/7 and mitochondrial membrane permeability assays. Modulation of the key genes involved in MAPK signaling pathway was also measured using immunoblotting array.
The inhibitory kinetic study has revealed that CHBC inhibited SNB19 and LN229 cell growth in a time-dependent manner. Furthermore, CHBC with the IC of 85 μM, mediated cell death through an apoptosis mechanism in both studied cell lines. The study also has revealed that CHBC targets GPR17 leading to the induction of apoptosis via the activation of Caspase 3/7 and dysfunction of mitochondrial membrane potential. In addition, CHBC treatment led to marked G2/M cell cycle arrest. The protein array has confirmed the anticancer effect of CHBC by the disruption of the mitogen-activated protein kinase pathway (MAPK).
Taken together, these results demonstrated that CHBC induced G2/M cell cycle arrest and apoptosis by disrupting MAPK signaling in human glioblastoma cells. This study concludes that CHBC represent a class of compounds for treating glioblastoma.
多形性胶质母细胞瘤（GBM）是最常见和最具侵袭性的成人原发性脑肿瘤。GBM 的特征是存在对化疗有抗性的异质细胞群体。最近，我们合成了 CHBC，这是一种针对 GBM 生物标志物 G 蛋白偶联受体 17 的新型吲哚衍生物，也是 GBM 细胞的抑制剂。在这项研究中，我们进一步研究了 CHBC 在分子水平上对 GBM 的治疗效率及其在诱导 GBM 细胞死亡中的潜在机制。
使用时间依赖性抑制剂测定法在Glioblastoma 细胞 LN229 和 SNB19 中测定 CHBC 和 TMZ 的作用。使用 PI 染色和图像分析测定药物诱导的细胞周期停滞。使用凋亡、caspase 3/7 和线粒体膜通透性测定研究 CHBC 诱导凋亡的作用及其作用机制。还使用免疫印迹分析阵列测量了 MAPK 信号通路中关键基因的调节。
抑制动力学研究表明，CHBC 以时间依赖性方式抑制 SNB19 和 LN229 细胞的生长。此外，在两种研究细胞系中，IC 为 85μM 的 CHBC 通过凋亡机制介导细胞死亡。该研究还表明，CHBC 靶向 GPR17，通过激活 Caspase 3/7 和线粒体膜电位功能障碍诱导凋亡。此外，CHBC 处理导致明显的 G2/M 细胞周期停滞。蛋白阵列通过破坏丝裂原活化蛋白激酶途径（MAPK）证实了 CHBC 的抗癌作用。
综上所述，这些结果表明 CHBC 通过破坏 MAPK 信号通路诱导人胶质母细胞瘤细胞的 G2/M 细胞周期停滞和凋亡。这项研究得出结论，CHBC 代表了一类用于治疗胶质母细胞瘤的化合物。
