Cytotoxicity, apoptosis, cell cycle arrest, reactive oxygen species, mitochondrial membrane potential, and Western blotting analysis of ruthenium(II) complexes

2013 ◽  
Vol 18 (8) ◽  
pp. 873-882 ◽  
Author(s):  
Gan-Jian Lin ◽  
Guang-Bin Jiang ◽  
Yang-Yin Xie ◽  
Hong-Liang Huang ◽  
Zhen-Hua Liang ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Cell Cycle ◽  
Membrane Potential ◽  
Cell Cycle Arrest ◽  
Mitochondrial Membrane Potential ◽  
Western Blotting ◽  
Mitochondrial Membrane ◽  
Oxygen Species ◽  
Western Blotting Analysis ◽  
Cycle Arrest

scholarly journals Effects of Berberine on Cell Cycle, DNA, Reactive Oxygen Species, and Apoptosis in L929 Murine Fibroblast Cells

2015 ◽  
Vol 2015 ◽  
pp. 1-13 ◽  
Author(s):  
Manman Gu ◽  
Jing Xu ◽  
Chunyang Han ◽  
Youxi Kang ◽  
Tengfei Liu ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Cell Cycle ◽  
Dna Damage ◽  
Membrane Potential ◽  
Mitochondrial Membrane Potential ◽  
Cell Apoptosis ◽  
Mitochondrial Membrane ◽  
Oxygen Species ◽  
Cycle Arrest ◽  
Murine Fibroblast

Berberine, an isoquinoline alkaloid isolated from several traditional Chinese herbal medicines (TCM), exhibits a strong antimicrobial activity in the treatment of diarrhea. However, it causes human as well as animal toxicity from heavy dosage. The present study was conducted to investigate the cytotoxicity of berberine and its possible trigger mechanisms resulting in cell cycle arrest, DNA damage, ROS (reactive oxygen species) level, mitochondrial membrane potential change, and cell apoptosis in L929 murine fibroblast (L929) cells. The cells were culturedin vitroand treated with different concentrations of berberine for 24 h. The results showed that cell viability was significantly decreased in a subjected dose-dependent state; berberine concentrations were higher than 0.05 mg/mL. Berberine at a concentration above 0.1 mg/mL altered the morphology of L929 cells. Cells at G2/M phase were clear that the level of ROS and cell apoptosis rates increased in 0.1 mg/mL group. Each DNA damage indicator score (DIS) increased in groups where concentration of berberine was above 0.025 mg/mL. The mitochondrial membrane potential counteractive balance mechanics were significantly altered when concentrations of berberine were above 0.005 mg/mL. In all, the present study suggested that berberine at high dosage exhibited cytotoxicity on L929 which was related to resultant: cell cycle arrest; DNA damage; accumulation of intracellular ROS; reduction of mitochondrial membrane potential; and cell apoptosis.

scholarly journals Pterostilbene Induces Cell Apoptosis and Cell Cycle Arrest in T-Cell Leukemia/Lymphoma by Suppressing the ERK1/2 Pathway

2017 ◽  
Vol 2017 ◽  
pp. 1-11 ◽  
Author(s):  
Gaomei Chang ◽  
Wenqin Xiao ◽  
Zhijian Xu ◽  
Dandan Yu ◽  
Bo Li ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Cell Cycle ◽  
Membrane Potential ◽  
T Cell ◽  
Cell Cycle Arrest ◽  
Mitochondrial Membrane ◽  
Oxygen Species ◽  
T Cell Leukemia ◽  
Cell Leukemia ◽  
Cycle Arrest

Pterostilbene is a natural 3,5-dimethoxy analog oftrans-resveratrol that has been reported to have antitumor, antioxidant, and anti-inflammatory effects. T-cell leukemia/lymphoma is one of the more aggressive yet uncommon non-Hodgkin lymphomas. Although there has been increasing research into T-cell leukemia/lymphoma, the molecular mechanisms of the antitumor effects of pterostilbene against this malignancy are still largely unknown. The aim of this study is to confirm the effects of pterostilbene in T-cell leukemia/lymphoma. Jurkat and Hut-78 cells treated with pterostilbene were evaluated for cell proliferation using Cell Counting Kit-8, and apoptosis, cell cycle progression, reactive oxygen species generation, and mitochondrial membrane potential were analyzed using flow cytometry. The level of protein expression was detected by western blot. The results demonstrated that pterostilbene significantly inhibited the growth of T-cell leukemia/lymphoma cell lines in vitro and induced apoptosis in a dose- and time-dependent manner. Moreover, pterostilbene treatment markedly induced S-phase cell cycle arrest, which was accompanied by downregulation of cdc25A, cyclin A2, and CDK2. Pterostilbene also induced the generation of reactive oxygen species and the loss of mitochondrial membrane potential and inhibited ERK1/2 phosphorylation. Taken together, our study demonstrated the potential of pterostilbene to be an effective treatment for T-cell leukemia/lymphoma.

Reserpine Induces Apoptosis and Cell Cycle Arrest in Hormone Independent Prostate Cancer Cells through Mitochondrial Membrane Potential Failure

2019 ◽  
Vol 18 (9) ◽  
pp. 1313-1322 ◽  
Author(s):  
Manjula Devi Ramamoorthy ◽  
Ashok Kumar ◽  
Mahesh Ayyavu ◽  
Kannan Narayanan Dhiraviam
Keyword(s):  
Prostate Cancer ◽  
Reactive Oxygen Species ◽  
Cell Cycle ◽  
Membrane Potential ◽  
Cancer Cells ◽  
Mitochondrial Membrane Potential ◽  
Mitochondrial Membrane ◽  
Reactive Oxygen ◽  
Oxygen Species ◽  
Prostate Cancer Cells

Background: Reserpine, an indole alkaloid commonly used for hypertension, is found in the roots of Rauwolfia serpentina. Although the root extract has been used for the treatment of cancer, the molecular mechanism of its anti-cancer activity on hormonal independent prostate cancer remains elusive. Methods: we evaluated the cytotoxicity of reserpine and other indole alkaloids, yohimbine and ajmaline on Prostate Cancer cells (PC3) using MTT assay. We investigated the mechanism of apoptosis using a combination of techniques including acridine orange/ethidium bromide staining, high content imaging of Annexin V-FITC staining, flow cytometric quantification of the mitochondrial membrane potential and Reactive Oxygen Species (ROS) and cell cycle analysis. Results: Our results indicate that reserpine inhibits DNA synthesis by arresting the cells at the G2 phase and showed all standard sequential features of apoptosis including, destabilization of mitochondrial membrane potential, reduced production of reactive oxygen species and DNA ladder formation. Our in silico analysis further confirmed that indeed reserpine docks to the catalytic cleft of anti-apoptotic proteins substantiating our results. Conclusion: Collectively, our findings suggest that reserpine can be a novel therapeutic agent for the treatment of androgen-independent prostate cancer.

Tet-Regulated Expression of Mutant Neutrophil Elastase in HL-60 Cells: A Cellular Model of Severe Congenital Neutropenia.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 1452-1452
Author(s):  
Andrew A.G. Aprikyan ◽  
Nara A. Markosyan
Keyword(s):  
Cell Cycle ◽  
Membrane Potential ◽  
Cell Cycle Arrest ◽  
Mitochondrial Membrane Potential ◽  
Neutrophil Elastase ◽  
Mitochondrial Membrane ◽  
Apoptotic Cell ◽  
Induced Expression ◽  
Maturation Arrest ◽  
Cycle Arrest

Abstract Severe congenital neutropenia (SCN) is an inheritable hematopoietic disorder characterized by extremely low level of circulating neutrophils, ineffective intramedulary hematopoiesis with “maturation arrest” at the promyelocytic stage of differentiation, recurrent severe infections, and evolution to acute myelogenous leukemia (AML). Accelerated apoptosis of bone marrow-derived myeloid progenitor cells and cell cycle arrest of CD34+ cells was reported in SCN patients. We also reported that heterozygous mutations in the neutrophil elastase (NE) gene have been identified in approximately 80% of SCN patients. Transient expression of mutant but not normal NE triggered accelerated apoptotic cell death of human promyelocytic HL-60 cells supporting the causative role of mutant NE in pathogenesis of SCN. Here we present cellular model of SCN based on newly established tet-off HL-60 human promyelocytic cell line with inducible doxycycline-regulated expression of mutant NE with 8-amino acid deletion identified in a patient evolved to develop AML. Induced expression of mutant elastase in tet-off HL-60 cells treated with DMSO led to a block of differentiation or “maturation arrest” at the promyelocytic stage of differentiation with a significantly reduced proportion of differentiated cells (approximately 20% vs 70% in control) and higher proportion (~40%) of primitive undifferentiated cells compared with control DMSO-treated HL-60 cells expressing only normal NE (~10%). Flow cytometry analysis of annexin V-labeled cells repeatedly revealed approximately 2-fold higher rate of apoptosis in tet-off HL-60 cells with induced expression of mutant NE compared with control cells. FACS analysis of DAPI-labeled tet-off HL-60 cells with induced expression of mutant NE revealed abnormal cell cycle progression with gradual accumulation and apparent arrest of cells in G1-phase of the cell cycle similar to that reported for SCN patients. Interestingly, the apoptotic cell shrinking and swelling as determined by flow cytometry was observed in all phases of the cell cycle suggesting that accelerated apoptosis rather than cell cycle arrest is the primary abnormality caused by expression of mutant NE, and the abnormal cell cycle progression is a consequence of this impaired cell survival. Further analysis revealed a dramatic reduction in mitochondrial membrane potential of tet-off HL-60 cells expressing mutant NE compared with control cells. The reduced mitochondrial membrane potential as determined by FACS was observed as early as 24 hours of induction of mutant NE expression and before the accelerated apoptosis or cell cycle arrest was detected. Western blot analysis demonstrated that caspase-3 was not activated in the cells even after 3 days of mutant NE induction. Further analysis of apoptosis regulators revealed a down-regulation of Bcl-2 expression and up-regulation of Bax in cells with induced expression of mutant elastase. These data suggest that mutant elastase-mediated “maturation arrest” of human promyelocytic cells in patients with SCN and SCN/AML is associated with reduced expression of anti-apoptotic Bcl-2 and upregulation of pro-apoptotic Bax proteins that trigger a dramatic reduction in mitochondrial membrane potential and subsequent caspase-independent apoptosis and cell cycle arrest. Current studies focused on elucidating and characterizing specific molecular interactions mediating the pro-apoptotic effect of mutant neutrophil elastase in SCN.

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