Antiproliferative, genotoxic and oxidant activities of cyclosativene in rat neuron and neuroblastoma cell lines

Cyclosativene (CSV) is a tetracyclic sesquiterpene found in the essential oils of Centaurea cineraria (Asteraceae) and Abies magnifica A. Murray (Pinaceae) plants. To the best of our knowledge, its cytotoxic, genotoxic and oxidant effects have never been studied on any cell lines. Therefore, we aimed to investigate the in vitro antiproliferative and/or cytotoxic properties, antioxidant/oxidant activity and genotoxic damage potential of CSV in healthy neurons and N2a neuroblastoma (N2a-NB) cell cultures. After treatment with 10-400 ?g/ml of CSV for 24 h, cell proliferation was measured by the MTT (3,(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The antioxidant activity was assessed by the total antioxidant capacity (TAC) and total oxidative stress (TOS) assays. To evaluate the level of DNA damage, single cell gel alkaline electrophoresis (SCGE) was used. The MTT assay showed that the application of CSV significantly reduced cell viability in both cell types. CSV treatments at higher doses led to decreases of TAC levels and increases of TOS levels in neuron and N2a-NB cells. The mean values of the total scores of cells showing DNA damage were not found to be significantly different from the control values in both cells. In conclusion, this study suggests that CSV has weak anticancer potential.

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Antioxidative, anticancer and genotoxic properties of α-pinene on N2a neuroblastoma cells

Biologia ◽

10.2478/s11756-013-0230-2 ◽

2013 ◽

Vol 68 (5) ◽

Cited By ~ 34

Author(s):

Elanur Aydin ◽

Hasan Türkez ◽

Fatime Geyikoğlu

Keyword(s):

Biological Activities ◽

Neuroblastoma Cells ◽

Cell Types ◽

Mean Values ◽

N2a Cells ◽

Diphenyltetrazolium Bromide ◽

Total Antioxidant ◽

Total Oxidative Stress ◽

Oxidative Effects

Abstractα-Pinene, an organic monoterpene, is found in essential oils of pine and coniferous trees. To date, although various biological activities of α-pinene have been demonstrated, its neurotoxicity has never been explored. Therefore in this study, we aimed to describe in vitro antiproliferative and/or cytotoxic properties by 3-(4,5-dimetylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test, genotoxic damage potentials by single cell gel electrophoresis, and oxidative effects by total antioxidant capacity (TAC) and total oxidative stress (TOS) analysis of α-pinene. Statistical analysis of MTT assay results indicated significant (p < 0.05) decreases of the cell proliferation rates in healthy neurons treated with α-pinene at only 400 mg/L, while significant decreases were observed in N2a cells at 100, 200 and 400 mg/L. On the other hand, the mean values of the total scores of cells showing DNA damage were not found significantly different from the control values on both cells. In addition, our results indicated that 10 and 25 mg/L of α-pinene treatment caused increases of TAC levels in primary rat neurons without any alterations of its level in N2a cells. However, α-pinene treatments at higher doses led to increases of TOS levels in both cell types. Overall our results suggest that α-pinene is of a limited therapeutic use as an anticancer agent.

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Cytotoxic and cytogenetic effects of α-copaene on rat neuron and N2a neuroblastoma cell lines

Biologia ◽

10.2478/s11756-014-0393-5 ◽

2014 ◽

Vol 69 (7) ◽

Cited By ~ 17

Author(s):

Hasan Turkez ◽

Basak Togar ◽

Abdulgani Tatar ◽

Fatime Geyıkoglu ◽

Ahmet Hacımuftuoglu

Keyword(s):

Cell Lines ◽

Cell Cultures ◽

Neuroblastoma Cell ◽

Medicinal And Aromatic Plants ◽

Damage Potential ◽

Total Antioxidant ◽

Total Oxidative Status ◽

Oxidative Effects ◽

First Time

AbstractAlpha-copaene (α-COP), a tricyclic sesquiterpene, is present in several essential oils of medicinal and aromatic plants and has antioxidant and antigenotoxic features. Its cytotoxic, cytogenetic and oxidative effects have not been investigated in neuron and N2a neuroblastoma (NB) cell cultures. Therefore, we aimed to describe in vitro: (i) cytotoxic properties by 3-(4,5-dimetylthiazol-2-yl)-2,5-diphenlytetrazolium bromide test; (ii) antioxidant/oxidant activity by total antioxidant capacity (TAC) and total oxidative status (TOS) analysis; and (iii) genotoxic damage potential by single cell gel electrophoresis — of α-COP in healthy neuron and N2a-NB cell cultures for the first time. Significant (P < 0.05) decrease in cell proliferation were observed in cultured primary rat neurons starting with the concentration of 150 mg/L and in N2a-NB cells starting with 100 mg/L. In addition, 25 mg/L of α-COP treatment caused increase of TAC levels and α-COP treatments at higher doses led to increase of TOS levels in neuron N2a-NB cell cultures. Moreover, none of the tested concentrations of α-COP have shown a genotoxic effect on both cell lines. Our findings clearly demonstrate that α-COP exhibited mild cytotoxic effects on N2a-NB cell line. In conclusion, α-COP may have potential as an anticancer agent, which needs to be further studied.

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Anticancer and Antioxidant Properties of Terpinolene in Rat Brain Cells

Archives of Industrial Hygiene and Toxicology ◽

10.2478/10004-1254-64-2013-2365 ◽

2013 ◽

Vol 64 (3) ◽

pp. 415-424 ◽

Cited By ~ 24

Author(s):

Elanur Aydin ◽

Hasan Türkez ◽

Şener Taşdemir

Keyword(s):

Biological Activities ◽

Antioxidant Properties ◽

Neuroblastoma Cells ◽

In Vitro Study ◽

Cell Types ◽

Damage Potential ◽

Total Antioxidant ◽

Antiproliferative Agent ◽

Total Oxidative Stress

Abstract Terpinolene (TPO) is a natural monoterpene present in essential oils of many aromatic plant species. Although various biological activities of TPO have been demonstrated, its neurotoxicity has never been explored. In this in vitro study we investigated TPO’s antiproliferative and/or cytotoxic properties using the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) test, genotoxic damage potential using the single-cell gel electrophoresis (SCGE), and oxidative effects through total antioxidant capacity (TAC) and total oxidative stress (TOS) in cultured primary rat neurons and N2a neuroblastoma cells. Dose-dependent effects of TPO (at 10 mg L-1, 25 mg L-1, 50 mg L-1, 100 mg L-1, 200 mg L-1, and 400 mg L-1) were tested in both cell types. Significant (P<0.05) decrease in cell proliferation were observed in cultured primary rat neurons starting with the dose of 100 mg L-1 and in N2a neuroblastoma cells starting with 50 mg L-1. TPO was not genotoxic in either cell type. In addition, TPO treatment at 10 mg L-1, 25 mg L-1, and 50 mg L-1 increased TAC in primary rat neurons, but not in N2a cells. However, at concentrations above 50 mg L-1 it increased TOS in both cell types. Our findings clearly demonstrate that TPO is a potent antiproliferative agent for brain tumour cells and may have potential as an anticancer agent, which needs to be further studied.

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In Vitro Cytotoxicity Induced by the Bufadienolides 1α,2α-Epoxyscillirosidine and Lanceotoxin B on Rat Myocardial and Mouse Neuroblastoma Cell Lines

Toxins ◽

10.3390/toxins11010014 ◽

2019 ◽

Vol 11 (1) ◽

pp. 14 ◽

Cited By ~ 2

Author(s):

Danielle Henn ◽

Annette Venter ◽

Christo Botha

Keyword(s):

Cell Lines ◽

Neuroblastoma Cell ◽

Cell Types ◽

Cell Ultrastructure ◽

In Vitro Cytotoxicity ◽

Cytoplasmic Material ◽

Mouse Neuroblastoma ◽

Transmission Electron ◽

Half Maximal Effective Concentration

Consumption of bufadienolide-containing plants are responsible for many livestock mortalities annually. Bufadienolides are divided into two groups; non-cumulative bufadienolides and cumulative bufadienolides. Cumulative bufadienolides are referred to as neurotoxic, as the chronic intoxication with this type of bufadienolide results in a paretic/paralytic syndrome known as ‘krimpsiekte’. The in vitro cytotoxicity of a non-cumulative bufadienolide, 1α,2α-epoxyscillirosidine, and a cumulative bufadienolide, lanceotoxin B, were compared using the MTT ((3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reduction) assay after exposing rat myocardial (H9c2) and mouse neuroblastoma (Neuro-2a) cell lines. The effect of these two bufadienolides on cell ultrastructure was also investigated using transmission electron microscopy (TEM). H9c2 cells exhibited greater cytotoxicity when exposed to 1α,2α-epoxyscillirosidine, compared to lanceotoxin B. In contrast, Neuro-2a cells were more susceptible to lanceotoxin B. The EC50 (half maximal effective concentration) of lanceotoxin B exposure of Neuro-2a cells for 24–72 h ranged from 4.4–5.5 µM compared to EC50s of 35.7–37.6 µM for 1α,2α-epoxyscillirosidine exposure of Neuro-2a cells over the same period. 1α,2α-Epoxyscillirosidine induced extensive vacuolization in both cell types, with swollen RER (rough endoplasmic reticulum) and perinuclear spaces. Lanceotoxin B caused swelling of the mitochondria and sequestration of cytoplasmic material within autophagic vesicles. These results corroborate the notion that cumulative bufadienolides are neurotoxic.

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The in vitro cytotoxicity, genotoxicity and oxidative damage potential of dapagliflozin, on cultured human blood cells

Turkish Journal of Biochemistry ◽

10.1515/tjb-2019-0111 ◽

2019 ◽

Vol 44 (5) ◽

pp. 692-698

Author(s):

Kenan Çadırcı ◽

Özlem Özdemir Tozlu ◽

Hasan Türkez

Keyword(s):

Human Blood ◽

Blood Cells ◽

In Vitro Cytotoxicity ◽

Damage Potential ◽

Human Blood Cells ◽

Diphenyltetrazolium Bromide ◽

Total Antioxidant ◽

Release Assay ◽

Total Oxidative Stress

Abstract Objectives Dapagliflozin (DAPA), is a potent SGLT-2 inhibitor for the treatment of patients with type 2 diabetes. DAPA has a good clinical and biological tolerance profile. However little information is available on its potential effects on cultured human blood cells. The evaluation of the in vitro cytotoxicity, genotoxicity potential and antioxidant/oxidant activity of DAPA in primary human whole blood cell cultures was aimed in this study. Materials and methods Cell viability was measured by the MTT [3,(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] and lactate dehydrogenase (LDH) leakage assays. The antioxidant/oxidant activity was determined by measuring the total antioxidant capacity (TAC) and total oxidative stress (TOS) levels. To assess the genotoxicity of DAPA, chromosomal aberration (CA) frequencies were determined. Results MTT and LDH release assay exhibited that exposure to different doses of DAPA did not changed significantly the proliferation of cells. The results of TAC and TOS assays were showed that TAC level was elevated while TOS level did not altered in DAPA-treated cells. Moreover, any increase in the frequency of CA did not found on cultures blood cells. Conclusion These data indicate that DAPA has not cytotoxic and genotoxic potential in cultured human blood cells, also, induces the increasing antioxidant activity.

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The Telomerase Inhibitor RHPS4 Induces Telomere Shortening, DNA-Damage and Cell Death in AML Cells and Chemosensitises Cells to Daunorubicin.

Blood ◽

10.1182/blood.v114.22.2760.2760 ◽

2009 ◽

Vol 114 (22) ◽

pp. 2760-2760

Author(s):

Monica Pallis ◽

Dotun Ojo ◽

Jaineeta Richardson ◽

John Ronan ◽

Malcolm Stevens ◽

...

Keyword(s):

Dna Damage ◽

Cell Death ◽

Telomere Length ◽

Progenitor Cells ◽

Cell Lines ◽

Dna Damage Response ◽

Cell Types ◽

Telomere Shortening ◽

Damage Response

Abstract Abstract 2760 Poster Board II-736 The quadruplex ligand RHPS4 is the lead compound in a drug discovery program at the University of Nottingham. It has been shown to bind to telomeres and inhibit telomerase, and subsequently induces growth arrest in progenitor cells from cancer cell lines whilst sparing normal haematopoietic progenitor cells. We explored its in vitro effects in AML cells, which are reported generally to have considerably shorter telomeres than normal CD34+ cells. AML cell lines were grown for 21 days in suspension culture. Primary samples were cultured for 14 days in semi-solid medium. Telomere length was measured by Southern blotting. γH2A.X was used to identify a DNA damage response, and cell viability was measured flow cytometrically with 7-amino actinomycin D. As reported in other tumour cell types, sensitivity to RHPS4 was found to be greatest in those AML cells with the shortest telomeres. In the OCI-AML3 cell line 0.3 μM RHPS4 inhibited cell growth by 50% in a 21 day clonogenic assay, accompanied by shortening of telomeres from 2.6 Kb to <1 Kb. Molm 13 cells (initial telomere length 3.2kB) also underwent telomere shortening in the presence of 0.3 μM RHPS4 (2.8Kb), whereas TF1a and U937 (both with initial telomere lengths approximately 6.5 kB) were insensitive at that concentration. After 6 days at 0.3 μM, RHPS4 was cytostatic, but at higher concentrations (1 μM) the drug was found to induce a substantial DNA damage response and loss of viability to OCI-AML3 cells. Moreover 0.3 μM RHPS4 enhanced the γH2A.X expression and cell death induced by the chemotherapy drug daunorubicin in these cells. Using 14 day clonogenic assays in primary AML samples (n=6), we found that the IC50 for RHPS4 alone was 0.7 μM. However, in the presence of 0.3 μM RHPS4, the median IC50 to daunorubicin was reduced from 19 nM to 5.5 nM. In conclusion we have determined that RHPS4 has telomere-shortening, cytostatic, cytotoxic and chemosensitising properties in AML cells. Disclosures: Stevens: Pharminox Ltd: director and shareholder of Pharminox Ltd which has a financial interest in RHPS4.

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Zingiberene attenuates hydrogen peroxide-induced toxicity in neuronal cells

Human & Experimental Toxicology ◽

10.1177/0960327114538987 ◽

2014 ◽

Vol 34 (2) ◽

pp. 135-144 ◽

Cited By ~ 6

Author(s):

B Togar ◽

H Türkez ◽

AD Stefano ◽

A Tatar ◽

D Cetin

Keyword(s):

Oxidative Stress ◽

Hydrogen Peroxide ◽

Mean Values ◽

Oxidative Damages ◽

Cortex Cell ◽

Diphenyltetrazolium Bromide ◽

Total Antioxidant ◽

Total Oxidative Stress ◽

First Time

In this experimental design, we explored the neuroprotective potential of zingiberene (ZGB), a monocyclic sesquiterpene, in hydrogen peroxide (H2O2)-induced toxicity in newborn rat cerebral cortex cell cultures for the first time. The rats were exposed to H2O2 for 6 h to determine the oxidative stress levels. To evaluate cell viability, both 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide and lactate dehydrogenase assays were carried out. Total antioxidant capacity (TAC) and total oxidative stress (TOS) parameters were used to evaluate oxidative changes. Besides determining 8-hydroxy-2-deoxyguanosine (8-OH-dG) levels in vitro, single-cell gel electrophoresis was also performed to measure the resistance of neuronal DNA to H2O2- exposed rats. Our results showed that survival and TAC levels of the cells decreased, while TOS, 8-OH-dG levels and the mean values of the total scores of cells showing DNA damage increased in the H2O2 alone-treated cultures. But pretreatment of ZGB suppressed the cytotoxicity, genotoxicity and oxidative stress that were increased by H2O2. Based on these observations, it is suggested that the sesquiterpene ZGB can be used as a novel and natural potential therapeutic in counteracting oxidative damages in the field of neurodegenerative disorders.

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Bestimmung der DNA-Schädigung in vitro

Nuklearmedizin ◽

10.1055/s-0038-1626526 ◽

2010 ◽

Vol 49 (S 01) ◽

pp. S64-S68

Author(s):

E. Dikomey

Keyword(s):

Dna Damage ◽

Cell Lines ◽

Chromosome Aberrations ◽

Genetic Factors ◽

Double Strand Breaks ◽

Strand Breaks ◽

Cell Inactivation ◽

Repair Mechanisms ◽

Break Repair

SummaryIonising irradiation acts primarily via induction of DNA damage, among which doublestrand breaks are the most important lesions. These lesions may lead to lethal chromosome aberrations, which are the main reason for cell inactivation. Double-strand breaks can be repaired by several different mechanisms. The regulation of these mechanisms appears be fairly different for normal and tumour cells. Among different cell lines capacity of doublestrand break repair varies by only few percents and is known to be determined mostly by genetic factors. Knowledge about doublestrand break repair mechanisms and their regulation is important for the optimal application of ionising irradiation in medicine.

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In Vitro Co-Exposure to CeO2 Nanomaterials from Diesel Engine Exhaust and Benzo(a)Pyrene Induces Additive DNA Damage in Sperm and Cumulus Cells but Not in Oocytes

Nanomaterials ◽

10.3390/nano11020478 ◽

2021 ◽

Vol 11 (2) ◽

pp. 478

Author(s):

Martina Cotena ◽

Mélanie Auffan ◽

Virginie Tassistro ◽

Noémie Resseguier ◽

Jérôme Rose ◽

...

Keyword(s):

Dna Damage ◽

Diesel Engine ◽

Fossil Fuels ◽

Cumulus Cells ◽

Ambient Air ◽

Cell Types ◽

Reproductive Organs ◽

Efficient System ◽

Polycyclic Aromatic

Benzo(a)pyrene (BaP) is a recognized reprotoxic compound and the most widely investigated polycyclic aromatic hydrocarbon in ambient air; it is widespread by the incomplete combustion of fossil fuels along with cerium dioxide nanomaterials (CeO2 NMs), which are used in nano-based diesel additives to decrease the emission of toxic compounds and to increase fuel economy. The toxicity of CeO2 NMs on reproductive organs and cells has also been shown. However, the effect of the combined interactions of BaP and CeO2 NMs on reproduction has not been investigated. Herein, human and rat gametes were exposed in vitro to combusted CeO2 NMs or BaP or CeO2 NMs and BaP in combination. CeO2 NMs were burned at 850 °C prior to mimicking their release after combustion in a diesel engine. We demonstrated significantly higher amounts of DNA damage after exposure to combusted CeO2 NMs (1 µg·L−1) or BaP (1.13 µmol·L−1) in all cell types considered compared to unexposed cells. Co-exposure to the CeO2 NMs-BaP mixture induced additive DNA damage in sperm and cumulus cells, whereas no additive effect was observed in rat oocytes. This result could be related to the structural protection of the oocyte by cumulus cells and to the oocyte’s efficient system to repair DNA damage compared to that of cumulus and sperm cells.

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Expression of the Metalloproteinase ADAM8 Is Upregulated in Liver Inflammation Models and Enhances Cytokine Release In Vitro

Mediators of Inflammation ◽

10.1155/2021/6665028 ◽

2021 ◽

Vol 2021 ◽

pp. 1-17

Author(s):

Tanzeela Awan ◽

Aaron Babendreyer ◽

Justyna Wozniak ◽

Abid Mahmood Alvi ◽

Viktor Sterzer ◽

...

Keyword(s):

Cell Lines ◽

Inflammatory Mediators ◽

Liver Tissue ◽

Stellate Cell ◽

Hepatoma Cell ◽

Cell Types ◽

Chronic Liver ◽

Liver Inflammation ◽

Tnf Α

Acute and chronic liver inflammation is driven by cytokine and chemokine release from various cell types in the liver. Here, we report that the induction of inflammatory mediators is associated with a yet undescribed upregulation of the metalloproteinase ADAM8 in different murine hepatitis models. We further show the importance of ADAM8 expression for the production of inflammatory mediators in cultured liver cells. As a model of acute inflammation, we investigated liver tissue from lipopolysaccharide- (LPS-) treated mice in which ADAM8 expression was markedly upregulated compared to control mice. In vitro, stimulation with LPS enhanced ADAM8 expression in murine and human endothelial and hepatoma cell lines as well as in primary murine hepatocytes. The enhanced ADAM8 expression was associated with an upregulation of TNF-α and IL-6 expression and release. Inhibition studies indicate that the cytokine response of hepatoma cells to LPS depends on the activity of ADAM8 and that signalling by TNF-α can contribute to these ADAM8-dependent effects. The role of ADAM8 was further confirmed with primary hepatocytes from ADAM8 knockout mice in which TNF-α and IL-6 induction and release were considerably attenuated. As a model of chronic liver injury, we studied liver tissue from mice undergoing high-fat diet-induced steatohepatitis and again observed upregulation of ADAM8 mRNA expression compared to healthy controls. In vitro, ADAM8 expression was upregulated in hepatoma, endothelial, and stellate cell lines by various mediators of steatohepatitis including fatty acid (linoleic-oleic acid), IL-1β, TNF-α, IFN-γ, and TGF-β. Upregulation of ADAM8 was associated with the induction and release of proinflammatory cytokines (TNF-α and IL-6) and chemokines (CX3CL1). Finally, knockdown of ADAM8 expression in all tested cell types attenuated the release of these mediators. Thus, ADAM8 is upregulated in acute and chronic liver inflammation and is able to promote inflammation by enhancing expression and release of inflammatory mediators.

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