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

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.

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Induction of Mitosis Delay And Apoptosis By RTA 404 In Glioblastoma Multiforme

10.21203/rs.3.rs-789618/v1 ◽

2021 ◽

Author(s):

Tai-Hsin Tsai ◽

Ann-Shung Lieu ◽

Tzuu-Yuan Huang ◽

Aij-Lie Kwan ◽

Chih-Lung Lin ◽

...

Keyword(s):

Cell Cycle ◽

Glioblastoma Multiforme ◽

Cell Apoptosis ◽

Cell Cycle Progression ◽

Structural Changes ◽

Cyclin B1 ◽

External Pressure ◽

Membrane Potential Change ◽

Cycle Progression

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, a trifluoroethylamidederivative of CDDO, is an Nrf2/ARE pathway activator. RTA 404 is used to inhibit proliferation and induce differentiation and apoptosis in glioma cells. However, it not clear what effect it may have on tumorigenesis in GBM.MethodsThis in vitro study evaluated the effects of RTA 404 on GBM cells. To do this, we treated GBM8401 cell lines with RTA 404 and assessed apoptosis, cell cycle. DNA content and induction of apoptosis were analyzed by flow cytometry and protein expression by Western blot analysis.ResultsRTA 404 significantly inhibited the proliferation induced cell apoptosis on GBM 8401 cell line. Typical plasma membrane undergoes structural changes that cause translocation of phosphatidylserine from the inside to outside. Due to cell external pressure cause mitochondrial membrane potential change lead to cell apoptosis. Caspase-3 active respond to apoptosis phenomenon, continuous progression of apoptosis.In addition, treatment with RTA 404 led to an accumulation of G2/M-phase cells. An analysis of Cyclin B1, CDK1 and Cyclin B1/CDK1 complex association suggested that cell cycle progression seems also to be regulated by RTA 404. Therefore, RTA 404 may not only induced cell cycle G2/M arrest, it may also exert apoptosis in established GBM cells. ConclusionRTA 404 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.

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(S)-10-Hydroxycamptothecin Inhibits Esophageal Squamous Cell Carcinoma Growth In Vitro and In Vivo Via Decreasing Topoisomerase I Enzyme Activity

Cancers ◽

10.3390/cancers11121964 ◽

2019 ◽

Vol 11 (12) ◽

pp. 1964 ◽

Cited By ~ 3

Author(s):

Mengqiu Song ◽

Shuying Yin ◽

Ran Zhao ◽

Kangdong Liu ◽

Joydeb Kumar Kundu ◽

...

Keyword(s):

Cell Cycle ◽

Enzymatic Activity ◽

Topoisomerase I ◽

Cell Cycle Progression ◽

Caspase 3 ◽

Cyclin B1 ◽

Phase Cell ◽

Cycle Progression

Topoisomerase (TOP) I plays a major role in the process of supercoiled DNA relaxation, thereby facilitating DNA replication and cell cycle progression. The expression and enzymatic activity of TOP I is positively correlated with tumor progression. Although the anticancer activity of (S)-10-Hydroxycamptothecin (HCPT), a TOP I specific inhibitor, has been reported in various cancers, the effect of HCPT on esophageal cancer is yet to be examined. In this study, we investigate the potential of HCPT to inhibit the growth of ESCC cells in vitro and verify its anti-tumor activity in vivo by using a patient-derived xenograft (PDX) tumor model in mice. Our study revealed the overexpression of TOP I in ESCC cells and treatment with HCPT inhibited TOP I enzymatic activity at 24 h and decreased expression at 48 h and 72 h. HCPT also induced DNA damage by increasing the expression of H2A.XS139. HCPT significantly decreased the proliferation and anchorage-independent growth of ESCC cells (KYSE410, KYSE510, KYSE30, and KYSE450). Mechanistically, HCPT inhibited the G2/M phase cell cycle transition, decreased the expression of cyclin B1, and elevated p21 expression. In addition, HCPT stimulated ESCC cells apoptosis, which was associated with elevated expression of cleaved PARP, cleaved caspase-3, cleaved caspase-7, Bax, Bim, and inhibition of Bcl-2 expression. HCPT dramatically suppressed PDX tumor growth and decreased the expression of Ki-67 and TOP I and increased the level of cleaved caspase-3 and H2A.XS139 expression. Taken together, our data suggested that HCPT inhibited ESCC growth, arrested cell cycle progression, and induced apoptosis both in vitro and in vivo via decreasing the expression and activity of TOP I enzyme.

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SKLB316, a novel small-molecule inhibitor of cell-cycle progression, induces G2/M phase arrest and apoptosis in vitro and inhibits tumor growth in vivo

Cancer Letters ◽

10.1016/j.canlet.2014.09.042 ◽

2014 ◽

Vol 355 (2) ◽

pp. 297-309 ◽

Cited By ~ 23

Author(s):

Yong Xia ◽

Qian Lei ◽

Yongxia Zhu ◽

Tinghong Ye ◽

Ningyu Wang ◽

...

Keyword(s):

Cell Cycle ◽

Tumor Growth ◽

Small Molecule ◽

Cell Cycle Progression ◽

Cycle Progression ◽

Phase Arrest ◽

Molecule Inhibitor ◽

M Phase

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Differential effects of UCN-01, staurosporine and CGP 41 251 on cell cycle progression and CDC2/cyclin B1 regulation in A431 cells synchronized at M phase by nocodazole

Anti-Cancer Drugs ◽

10.1097/00001813-199901000-00009 ◽

1999 ◽

Vol 10 (1) ◽

pp. 67-78 ◽

Cited By ~ 27

Author(s):

Tadakazu Akiyama ◽

Makiko Shimizu ◽

Masami Okabe ◽

Tatsuya Tamaoki ◽

Shiro Akinaga

Keyword(s):

Cell Cycle ◽

Cell Cycle Progression ◽

Cyclin B1 ◽

A431 Cells ◽

Cycle Progression ◽

Differential Effects ◽

M Phase

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Tryprostatin A, a specific and novel inhibitor of microtubule assembly

Biochemical Journal ◽

10.1042/bj3330543 ◽

1998 ◽

Vol 333 (3) ◽

pp. 543-548 ◽

Cited By ~ 152

Author(s):

Takeo USUI ◽

Masuo KONDOH ◽

Cheng-Bin CUI ◽

Tadanori MAYUMI ◽

Hiroyuki OSADA

Keyword(s):

Cell Cycle ◽

Cell Growth ◽

Cell Cycle Progression ◽

Microtubule Assembly ◽

Inhibition Mechanism ◽

Dependent Manner ◽

Cycle Progression ◽

Terminal Domain ◽

M Phase

We have investigated the cell cycle inhibition mechanism and primary target of tryprostatin A (TPS-A) purified from Aspergillus fumigatus. TPS-A inhibited cell cycle progression of asynchronously cultured 3Y1 cells in the M phase in a dose- and time-dependent manner. In contrast, TPS-B (the demethoxy analogue of TPS-A) showed cell-cycle non-specific inhibition on cell growth even though it inhibited cell growth at lower concentrations than TPS-A. TPS-A treatment induced the reversible disruption of the cytoplasmic microtubules of 3Y1 cells as observed by indirect immunofluorescence microscopy in the range of concentrations that specifically inhibited M-phase progression. TPS-A inhibited the assembly in vitro of microtubules purified from bovine brains (40% inhibition at 250 µM); however, there was little or no effect on the self-assembly of purified tubulin when polymerization was induced by glutamate even at 250 µM TPS-A. TPS-A did not inhibit assembly promoted by taxol or by digestion of the C-terminal domain of tubulin. However, TPS-A blocked the tubulin assembly induced by inducers interacting with the C-terminal domain, microtubule-associated protein 2 (MAP2), tau and poly-(l-lysine). These results indicate that TPS-A is a novel inhibitor of MAP-dependent microtubule assembly and, through the disruption of the microtubule spindle, specifically inhibits cell cycle progression at the M phase.

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Overproduction of Human Myt1 Kinase Induces a G2 Cell Cycle Delay by Interfering with the Intracellular Trafficking of Cdc2-Cyclin B1 Complexes

Molecular and Cellular Biology ◽

10.1128/mcb.19.7.5113 ◽

1999 ◽

Vol 19 (7) ◽

pp. 5113-5123 ◽

Cited By ~ 80

Author(s):

Feng Liu ◽

Cynthia Rothblum-Oviatt ◽

Christine E. Ryan ◽

Helen Piwnica-Worms

Keyword(s):

Cell Cycle ◽

Intracellular Trafficking ◽

Cell Cycle Progression ◽

Cyclin B1 ◽

Cyclin B ◽

Interaction Domain ◽

Cycle Progression ◽

Cell Cycle Delay ◽

Mutant Forms

ABSTRACT The Myt1 protein kinase functions to negatively regulate Cdc2-cyclin B complexes by phosphorylating Cdc2 on threonine 14 and tyrosine 15. Throughout interphase, human Myt1 localizes to the endoplasmic reticulum and Golgi complex, whereas Cdc2-cyclin B1 complexes shuttle between the nucleus and the cytoplasm. Here we report that overproduction of either kinase-active or kinase-inactive forms of Myt1 blocked the nuclear-cytoplasmic shuttling of cyclin B1 and caused cells to delay in the G2 phase of the cell cycle. The COOH-terminal 63 amino acids of Myt1 were identified as a Cdc2-cyclin B1 interaction domain. Myt1 mutants lacking this domain no longer bound cyclin B1 and did not efficiently phosphorylate Cdc2-cyclin B1 complexes in vitro. In addition, cells overproducing mutant forms of Myt1 lacking the interaction domain exhibited normal trafficking of cyclin B1 and unperturbed cell cycle progression. These results suggest that the docking of Cdc2-cyclin B1 complexes to the COOH terminus of Myt1 facilitates the phosphorylation of Cdc2 by Myt1 and that overproduction of Myt1 perturbs cell cycle progression by sequestering Cdc2-cyclin B1 complexes in the cytoplasm.

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NIPA Phosphorylation and Inactivation at G2/M Is Mediated by ERK2.

Blood ◽

10.1182/blood.v114.22.2513.2513 ◽

2009 ◽

Vol 114 (22) ◽

pp. 2513-2513

Author(s):

Michael Zech ◽

Anna Lena Illert ◽

Corinna Albers ◽

Florian Bassermann ◽

Christian Peschel ◽

...

Keyword(s):

Cell Cycle ◽

Cell Cycle Progression ◽

High Efficiency ◽

Conflicts Of Interest ◽

Cyclin B1 ◽

Cell Cycle Checkpoint ◽

Critical Event ◽

Checkpoint Control ◽

Cycle Progression

Abstract Abstract 2513 Poster Board II-490 Regulated oscillation of protein expression is an essential mechanism of cell cycle control. The SCF class of E3 ubiquitin ligases is involved in this process by targeting cell cycle regulatory proteins for degradation by the proteasome. We previously reported cloning of NIPA (Nuclear Interaction Partner of ALK) in complex with constitutively active oncogenic fusions of ALK, contributing to the development of lymphomas and sarcomas. Subsequently we characterized NIPA as a F-Box protein that defines an oscillating ubiquitin E3-ligase. The SCF-NIPA complex targets nuclear cyclin B1 for ubiquitination in interphase while phosphorylation of NIPA in late G2 phase and mitosis inactivates the complex to allow for accumulation of cyclin B1, a critical event for proper G2/M transition. Thus, SCF-NIPA executes an important G2/M checkpoint control. We recently specified three serine residues Ser 354, 359 and 395 implicated in NIPA phosphorylation at G2/M. These data suggest a sequential NIPA phosphorylation, where initial Ser 354 and 359 phosphorylation is most crucial for SCF-NIPA inactivation by dissociating the SCF-NIPA complex. Here we aimed to find the kinase responsible in this initial most important phosphorylation step. Using in vitro kinase assays we identified both ERK1 and ERK2 to phosphorylate NIPA with high efficiency. Mutation of either Ser 354 or Ser 359 abolished ERK-dependent NIPA phosphorylation. Inhibition of ERK1/2 activity in cell lines by specific inhibitors resulted in decreased NIPA phosphorylation at G2/M. To differentiate between phosphorylation by ERK1 and ERK2, we combined cell cycle analysis with stable expression of microRNA's targeting both isoforms. To this end NIH/3T3 cells were retrovirally transduced with microRNAs targeting ERK1 and 2 and cell cycle progression was analysed by BRDU/PI labeling. Using this approach, we are able to show that ERK2 but not ERK1 mediates NIPA phosphorylation at G2/M. Furthermore, ERK2 silencing leads to a distinct phenotype in cell cycle progression with a delay of ERK2 knockdown cells at the G2/M transition. Thus, our data indicate, that the recently described divergent functions of ERK1 and ERK2 in cell cycle regulation could be in part due to the differential ability of these kinases to phosphorylate and inactivate NIPA at G2/M. Since checkpoint proteins such as NIPA are constitutively inactivated in tumor cells ERK2 might represent an interesting target to reconstitute important cell cycle checkpoint controls in malignant cells. Disclosures: No relevant conflicts of interest to declare.

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LMNB2 promotes the progression of colorectal cancer by silencing p21 expression

Cell Death and Disease ◽

10.1038/s41419-021-03602-1 ◽

2021 ◽

Vol 12 (4) ◽

Author(s):

Chen-Hua Dong ◽

Tao Jiang ◽

Hang Yin ◽

Hu Song ◽

Yi Zhang ◽

...

Keyword(s):

Colorectal Cancer ◽

Cell Cycle ◽

Cell Proliferation ◽

Cell Cycle Progression ◽

Molecular Mechanisms ◽

Gene Set Enrichment Analysis ◽

Molecular Therapy ◽

Cycle Progression ◽

Cancer Tissues

AbstractColorectal cancer is the second common cause of death worldwide. Lamin B2 (LMNB2) is involved in chromatin remodeling and the rupture and reorganization of nuclear membrane during mitosis, which is necessary for eukaryotic cell proliferation. However, the role of LMNB2 in colorectal cancer (CRC) is poorly understood. This study explored the biological functions of LMNB2 in the progression of colorectal cancer and explored the possible molecular mechanisms. We found that LMNB2 was significantly upregulated in primary colorectal cancer tissues and cell lines, compared with paired non-cancerous tissues and normal colorectal epithelium. The high expression of LMNB2 in colorectal cancer tissues is significantly related to the clinicopathological characteristics of the patients and the shorter overall and disease-free cumulative survival. Functional analysis, including CCK8 cell proliferation test, EdU proliferation test, colony formation analysis, nude mouse xenograft, cell cycle, and apoptosis analysis showed that LMNB2 significantly promotes cell proliferation by promoting cell cycle progression in vivo and in vitro. In addition, gene set enrichment analysis, luciferase report analysis, and CHIP analysis showed that LMNB2 promotes cell proliferation by regulating the p21 promoter, whereas LMNB2 has no effect on cell apoptosis. In summary, these findings not only indicate that LMNB2 promotes the proliferation of colorectal cancer by regulating p21-mediated cell cycle progression, but also suggest the potential value of LMNB2 as a clinical prognostic marker and molecular therapy target.

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