Induction of Mitosis Delay and Apoptosis by CDDO-TFEA in Glioblastoma Multiforme

Tai-Hsin Tsai; Ann-Shung Lieu; Tzuu-Yuan Huang; Aij-Lie Kwan; Chih-Lung Lin; Yi-Chiang Hsu

doi:10.3389/fphar.2021.756228

Induction of Mitosis Delay and Apoptosis by CDDO-TFEA in Glioblastoma Multiforme

Front Pharmacol. 2021 Nov 8:12:756228. doi: 10.3389/fphar.2021.756228. eCollection 2021.

Authors

Tai-Hsin Tsai^{1

2

3}, Ann-Shung Lieu^{1

2

3}, Tzuu-Yuan Huang⁴, Aij-Lie Kwan^{1

2

3}, Chih-Lung Lin^{1

2

3}, Yi-Chiang Hsu⁵

Affiliations

¹ Division of Neurosurgery, Department of Surgery, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan.
² Department of Surgery, School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
³ Graduate Institutes of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
⁴ Department of Neurosurgery, Changhua Christian Hospital, Changhua, Taiwan.
⁵ School of Medicine, I-Shou University, Kaohsiung, Taiwan.

Abstract

Background: 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. Methods: This in vitro 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. Results: 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 G₂/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. Conclusion: CDDO-TFEA can inhibit proliferation, cell cycle progression and induce apoptosis in GBM cells in vitro, possibly though its inhibition of Cyclin B1, CDK1 expression, and Cyclin B1/CDK1 association and the promotion of CHK1 and CHK2 expression.

Keywords: CDDO-TFEA; GBM; RTA 404; apoptosis; cell cycle.