Shahcheraghi Seyed H, Lotfi Marzieh, Soukhtanloo Mohammad, Ghayour Mobarhan Majid, Jaliani Hossein Z, Sadeghnia Hamid R, Ghorbani Ahmad
Department of Medical Genetics and Molecular Medicine, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
Department of Medical Genetics, School of Medicine, Shahid Sadoughi University of medical sciences, Yazd, Iran.
Curr Mol Pharmacol. 2021;14(1):79-87. doi: 10.2174/1874467213666200512075507.
Glioblastoma is one of the most aggressive tumors of the central nervous system. Galbanic acid, a natural sesquiterpene coumarin, has shown favorable effects on cancerous cells in previous studies.
The aim of the present work was to evaluate the effects of galbanic acid on proliferation, migration, and apoptosis of the human malignant glioblastoma (U87) cells.
The anti-proliferative activity of the compound was determined by the MTT assay. Cell cycle alterations and apoptosis were analyzed via flow cytometry. Action on cell migration was evaluated by scratch assay and gelatin zymography. Quantitative Real-Time PCR was used to determine the expression of genes involved in cell migration (matrix metalloproteinases, MMPs) and survival (the pathways of PI3K/Akt/mTOR and WNT/β-catenin). Alteration in the level of protein Akt was determined by Western blotting.
Galbanic acid significantly decreased cell proliferation, inhibited cell cycle, and stimulated apoptosis of the glioblastoma cells. Moreover, it could decrease the migration capability of glioblastoma cells, which was accompanied by inhibition in the activity and expression of MMP2 and MMP9. While galbanic acid reduced the gene expression of Akt, mTOR, and PI3K and increased the PTEN expression, it had no significant effect on WNT, β-catenin, and APC genes. In addition, the protein level of p-Akt decreased after treatment with galbanic acid. The effects of galbanic acid were observed at concentrations lower than those of temozolomide.
Galbanic acid decreased proliferation, cell cycle progression, and survival of glioblastoma cells through inhibiting the PI3K/Akt/mTOR pathway. This compound also reduced the migration capability of the cells by suppressing the activity and expression of MMPs.
胶质母细胞瘤是中枢神经系统中最具侵袭性的肿瘤之一。没药酸是一种天然倍半萜香豆素，在先前的研究中已显示出对癌细胞有良好的作用。
本研究旨在评估没药酸对人恶性胶质母细胞瘤（U87）细胞增殖、迁移和凋亡的影响。
通过MTT法测定该化合物的抗增殖活性。通过流式细胞术分析细胞周期变化和凋亡情况。通过划痕试验和明胶酶谱法评估对细胞迁移的作用。采用定量实时PCR测定参与细胞迁移（基质金属蛋白酶，MMPs）和存活（PI3K/Akt/mTOR和WNT/β-连环蛋白通路）的基因表达。通过蛋白质印迹法测定蛋白Akt水平的变化。
没药酸显著降低胶质母细胞瘤细胞的增殖，抑制细胞周期，并刺激其凋亡。此外，它可以降低胶质母细胞瘤细胞的迁移能力，同时伴随着MMP2和MMP9活性及表达的抑制。没药酸降低了Akt、mTOR和PI3K的基因表达并增加了PTEN的表达，而对WNT、β-连环蛋白和APC基因无显著影响。此外，没药酸处理后p-Akt的蛋白水平降低。在低于替莫唑胺的浓度下观察到了没药酸的作用。
没药酸通过抑制PI3K/Akt/mTOR通路降低胶质母细胞瘤细胞的增殖、细胞周期进程和存活。该化合物还通过抑制MMPs的活性和表达降低了细胞的迁移能力。
