Tsai Tai-Hsin, Lieu Ann-Shung, Huang Tzuu-Yuan, Kwan Aij-Lie, Lin Chih-Lung, Hsu Yi-Chiang
Division of Neurosurgery, Department of Surgery, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan.
Department of Surgery, School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
Front Pharmacol. 2021 Nov 8;12:756228. doi: 10.3389/fphar.2021.756228. eCollection 2021.
Glioblastoma multiforme (GBM) is the vicious malignant brain tumor in adults. Despite advances multi-disciplinary treatment, GBM constinues to have a poor overall survival. CDDO-trifluoroethyl-amide (CDDO-TEFA), a trifluoroethylamidederivative of CDDO, is an Nrf2/ARE pathway activator. CDDO-TEFEA is used to inhibit proliferation and induce apoptosis in glioma cells. However, it not clear what effect it may have on tumorigenesis in GBM. This study evaluated the effects of CDDO-TFEA on GBM cells. To do this, we treated GBM8401 cell lines with CDDO-TFEA and assessed apoptosis, cell cycle. DNA content and induction of apoptosis were analyzed by flow cytometry and protein expression by Western blot analysis. CDDO-TFEA significantly inhibited the cell viability and induced cell apoptosis on GBM 8401 cell line. The annexin-FITC/PI assay revealed significant changes in the percentage of apoptotic cells. Treatment with CDDO-TFEA led to a significant reduction in the GBM8401 cells' mitochondrial membrane potential. A significant rise in the percentage of caspase-3 activity was detected in the treated cells. In addition, treatment with CDDO-TFEA led to an accumulation of G/M-phase cells. In addition, these results suggest that regarding increased protein synthesis during mitosis in the MPM-2 staining, indicative of a delay in the G2 checkpoint. An analysis of Cyclin B1, CDK1, Cyclin B1/CDK1 complex and CHK1 and CHK2 expression suggested that cell cycle progression seems also to be regulated by CDDO-TFEA. Therefore, CDDO-TFEA may not only induce cell cycle G2/M arrest, it may also exert apoptosis in established GBM cells. CDDO-TFEA can inhibit proliferation, cell cycle progression and induce apoptosis in GBM cells , possibly though its inhibition of Cyclin B1, CDK1 expression, and Cyclin B1/CDK1 association and the promotion of CHK1 and CHK2 expression.
多形性胶质母细胞瘤（GBM）是成人中恶性程度很高的脑肿瘤。尽管多学科治疗取得了进展，但GBM的总体生存率仍然很低。CDDO-三氟乙酰胺（CDDO-TEFA）是CDDO的三氟乙酰胺衍生物，是一种Nrf2/ARE途径激活剂。CDDO-TEFEA用于抑制胶质瘤细胞的增殖并诱导其凋亡。然而，尚不清楚它对GBM的肿瘤发生可能有什么影响。本研究评估了CDDO-TFEA对GBM细胞的影响。为此，我们用CDDO-TFEA处理GBM8401细胞系，并评估细胞凋亡、细胞周期。通过流式细胞术分析DNA含量和凋亡诱导情况，通过蛋白质印迹分析评估蛋白质表达。CDDO-TFEA显著抑制GBM 8401细胞系的细胞活力并诱导细胞凋亡。膜联蛋白-FITC/PI检测显示凋亡细胞百分比有显著变化。用CDDO-TFEA处理导致GBM8401细胞的线粒体膜电位显著降低。在处理过的细胞中检测到caspase-3活性百分比显著升高。此外，用CDDO-TFEA处理导致G/M期细胞积累。此外，这些结果表明，在MPM-2染色中，有丝分裂期间蛋白质合成增加，这表明G2期检查点延迟。对细胞周期蛋白B1、细胞周期蛋白依赖性激酶1、细胞周期蛋白B1/细胞周期蛋白依赖性激酶1复合物以及CHK1和CHK2表达的分析表明，细胞周期进程似乎也受CDDO-TFEA调控。因此，CDDO-TFEA不仅可能诱导细胞周期G2/M期阻滞，还可能在已建立的GBM细胞中诱导凋亡。CDDO-TFEA可以抑制GBM细胞的增殖、细胞周期进程并诱导其凋亡，可能是通过抑制细胞周期蛋白B1、细胞周期蛋白依赖性激酶1的表达以及细胞周期蛋白B1/细胞周期蛋白依赖性激酶1的结合，并促进CHK1和CHK2的表达来实现的。
