Cancer Biology Research Center, Cancer Institute of I.R. Iran, Tehran University of Medical Sciences, Tehran, Iran.
Department of Pharmacology, School of Medicine, Tehran University of Medical Sciences, P.O. Box 13145-784, Tehran, Iran.
Cancer Chemother Pharmacol. 2021 Jun;87(6):827-842. doi: 10.1007/s00280-021-04242-0. Epub 2021 Mar 10.
Glioblastoma multiforme (GBM) is one of the most aggressive human cancers. The c-MET receptor tyrosine kinase (RTK) which is frequently deregulated in GBM is considered as a promising target for GBM treatment. The c-MET plays a key role in cell proliferation, cell cycle progression, invasion, angiogenesis, and metastasis. Here, we investigated the anti-tumour activity of foretinib, a c-MET inhibitor, on three human GBM cells (T98G, U87MG and U251).
Anti-proliferative effect of foretinib was determined using MTT, crystal violet staining, and clonogenic assays. PI and Annexin V/PI staining flow cytometry were used to evaluate the effects of foretinib on cell cycle and apoptosis, respectively. Scratch assay, qRT-PCR, western blot, and zymography analyses were applied to elucidate the molecular mechanisms underlying the anti-tumour activity of foretinib.
Foretinib treatment reduced phosphorylation of c-MET on T98G and U251 cells, but not in U87MG cells. The highest inhibitory effect was observed in T98G cells (IC = 4.66 ± 0.29 µM) and the lowest one in U87MG cells (IC = 29.99 ± 1.31 µM). The results showed that foretinib inhibited the proliferation of GBM cells through a G2/M cell cycle arrest and mitochondrial-mediated apoptosis in association with alternation in expression of the related genes and protein-regulated G2/M phase and apoptosis. Foretinib diminished GBM cell invasion through downregulation of the proteolytic cascade of MMP2, uPA and uPAR and epithelial-mesenchymal transition (EMT)-related genes. A different GBM cell sensitivity pattern was noticeable in all experiments which demonstrated T98G as a sensitive and U87MG as a resistant phenotype to foretinib treatment.
The results indicated that foretinib might have the therapeutic potential against human GBM which deserve further investigation.
多形性胶质母细胞瘤（GBM）是最具侵袭性的人类癌症之一。在 GBM 中经常失调的 c-MET 受体酪氨酸激酶（RTK）被认为是 GBM 治疗的一个有前途的靶点。c-MET 在细胞增殖、细胞周期进程、侵袭、血管生成和转移中发挥关键作用。在这里，我们研究了 c-MET 抑制剂 foretinib 对三种人类 GBM 细胞（T98G、U87MG 和 U251）的抗肿瘤活性。
使用 MTT、结晶紫染色和集落形成测定法确定 foretinib 的抗增殖作用。PI 和 Annexin V/PI 染色流式细胞术分别用于评估 foretinib 对细胞周期和细胞凋亡的影响。划痕试验、qRT-PCR、western blot 和明胶酶谱分析用于阐明 foretinib 抗肿瘤活性的分子机制。
Foretinib 处理降低了 T98G 和 U251 细胞上 c-MET 的磷酸化，但在 U87MG 细胞中没有。在 T98G 细胞中观察到最高的抑制作用（IC = 4.66±0.29μM），而在 U87MG 细胞中观察到最低的抑制作用（IC = 29.99±1.31μM）。结果表明，foretinib 通过 G2/M 细胞周期阻滞和线粒体介导的凋亡抑制 GBM 细胞增殖，与相关基因和蛋白表达的改变以及 G2/M 期和凋亡的调节有关。Foretinib 通过下调 MMP2、uPA 和 uPAR 的蛋白水解级联以及上皮-间充质转化（EMT）相关基因来减少 GBM 细胞的侵袭。在所有实验中都观察到不同的 GBM 细胞敏感性模式，表明 T98G 对 foretinib 治疗敏感，而 U87MG 对 foretinib 治疗耐药。
结果表明，foretinib 可能对人类 GBM 具有治疗潜力，值得进一步研究。
