Transition metal-induced apoptosis in lymphocytes via hydroxyl radical generation, mitochondria dysfunction, and caspase-3 activation: an in vitro model for neurodegeneration

2004 ◽  
Vol 35 (3) ◽  
pp. 185-193 ◽  
Author(s):  
Marlene Jiménez Del Rı́o ◽  
Carlos Vélez-Pardo
Keyword(s):  
Transition Metal ◽  
Hydroxyl Radical ◽  
In Vitro Model ◽  
Caspase 3 ◽  
Mitochondria Dysfunction ◽  
Radical Generation ◽  
Vitro Model ◽  
Induced Apoptosis

Biochemical Characterisation of an In Vitro Model of Hepatocellular Apoptotic Cell Death

2009 ◽  
Vol 37 (2) ◽  
pp. 209-218 ◽  
Author(s):  
Mathieu Vinken ◽  
Elke Decrock ◽  
Elke De Vuyst ◽  
Luc Leybaert ◽  
Tamara Vanhaecke ◽  
...  
Keyword(s):  
Cell Death ◽  
In Vitro Model ◽  
Caspase 3 ◽  
Fas Ligand ◽  
Apoptotic Cell ◽  
Annexin V ◽  
Apoptotic Cell Death ◽  
Biochemical Characterisation ◽  
Vitro Model

This study was set up to critically evaluate a commonly-used in vitro model of hepatocellular apoptotic cell death, in which freshly isolated hepatocytes, cultured in a monolayer configuration, are exposed to a combination of Fas ligand and cycloheximide for six hours. A set of well-acknowledged cell death markers was addressed: a) cell morphology was studied by light microscopy; b) apoptotic and necrotic cell populations were quantified by in situ staining with Annexin-V, Hoechst 33342 and propidium iodide (PI); c) apoptotic and necrotic activities were monitored by probing caspase 3-like activity and measuring the extracellular leakage of lactate dehydrogenase (LDH), respectively; and d) the expression of apoptosis regulators was investigated by immunoblotting. The initiation of apoptosis was evidenced by the activation of caspase 8 and caspase 9, and increased Annexin-V reactivity. Progression through the apoptotic process was confirmed by the activation of caspase 3 and Bid, the enhanced expression of Bax, and the occurrence of nuclear fragmentation. Late transition to a necrotic appearance was demonstrated by an increased number of PI-positive cells and augmented extracellular release of LDH. Thus, the in vitro model allows the study of the entire course of Fas-mediated hepatocellular apoptotic cell death, which is not possible in vivo. This experimental system can serve a broad range of in vitro pharmaco-toxicological purposes, thereby directly assisting in the reduction of animal experimentation.

The Activity of Urolithin A and M4 Valerolactone, Colonic Microbiota Metabolites of Polyphenols, in a Prostate Cancer In Vitro Model

Planta Medica ◽  
2018 ◽  
Vol 85 (02) ◽  
pp. 118-125 ◽  
Author(s):  
Iwona Stanisławska ◽  
Sebastian Granica ◽  
Jakub Piwowarski ◽  
Joanna Szawkało ◽  
Krzysztof Wiązecki ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
In Vitro Model ◽  
Specific Antigen ◽  
Lncap Cells ◽  
Isobolographic Analysis ◽  
Human System ◽  
Urolithin A ◽  
Vitro Model ◽  
Induced Apoptosis

AbstractThe gut microbiota-derived metabolites of ellagitannins and green tea catechins, urolithin A (uroA) and 5-(3′,4′,5′-trihydroxyphenyl)-γ-valerolactone (M4), respectively, are among the main compounds absorbed into human system after ingestion of these polyphenols. The aim of this study was to establish the effects of M4, uroA, and their combinations on LNCaP cells, an androgen dependent prostate cancer in vitro model.. The LNCaP cells were incubated with increasing concentrations of tested metabolites. The cell proliferation was determined by measurement of DNA-bisbenzimide H 33 258 complexes fluorescence. The isobolographic analysis was used to establish the type of interaction between metabolites. The apoptosis, androgen receptor (AR) localization, and phosphorylation of Akt kinase were measured by flow cytometry. Prostate-specific antigen (PSA) secretion was determined by ELISA. M4 showed modest antiproliferative activity in LNCaP cells (IC50 = 117 µM; CI: 81 – 154). UroA decreased proliferation (IC50 = 32.7 µM; CI: 24.3 – 41.1) and induced apoptosis of LNCaP cells. The mixture of M4 with uroA had synergistic antiproliferative effect. Moreover, M4 potentiated inhibition of PSA secretion and enhanced retention of AR in cytoplasm caused by uroA. Interestingly, uroA increased levels of pSer473 Akt in LNCaP cells. These results show that colonic metabolites may contribute to chemoprevention of prostate cancer by varied polyphenol-rich diet or composite polyphenol preparations.

Development and initial application of an in vitro model of apoptosis in rodent cholangiocytes

1997 ◽  
Vol 272 (1) ◽  
pp. G106-G115 ◽  
Author(s):  
F. G. Que ◽  
G. J. Gores ◽  
N. F. LaRusso
Keyword(s):  
In Vitro Model ◽  
Interleukin 1 ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Initial Application ◽  
Histological Data ◽  
Vitro Model ◽  
Induced Apoptosis ◽  
Intracellular Ions

Although histological data suggest that cholangiocytes die by apoptosis in human liver diseases, no information exists on the mechanisms of cholangiocyte apoptosis. Thus our aims were to establish an in vitro model of cholangiocyte apoptosis and to test the hypothesis that changes in intracellular ions would cause apoptosis in cholangiocytes by a protease-sensitive pathway. A large number of proapoptotic agents were ineffective in inducing apoptosis in rat or human cholangiocytes in culture; in contrast, beauvericin, a K+ ionophore, caused apoptosis in both cell lines, despite their expression of Bcl-2. Although beauvericin decreased intracellular K+ and increased intracellular Ca2+, abolishing the K+ gradient did not prevent beauvericin-induced apoptosis; in contrast, omission of extracellular Ca2+ inhibited apoptosis by 42%. The interleukin-1 beta-converting enzyme (ICE) family protease inhibitor, Z-Val-Ala-Asp chloromethylketone, inhibited apoptosis in a concentration-dependent manner. By Northern blot analysis, cholangiocytes expressed the mRNA for three members of the ICE protease family: ICE, ICE/ CED-3 homologue-1 (ICH-1), and cysteine protease P-32 (CPP-32). Cleavage of a substrate for CPP-32-like protease activity, but not a substrate for ICE and ICH-1, increased after beauvericin treatment. In summary, we have established an in vitro model of apoptosis in cholangiocytes. Our data suggest that beauvericin-induced apoptosis occurs by a Ca(2+)-dependent CPP-32 protease-sensitive pathway despite cholangiocyte expression of Bcl-2.

scholarly journals A novel methylated analogue of L-Mimosine exerts its therapeutic potency through ROS production and ceramide-induced apoptosis in malignant melanoma

2021 ◽  
Author(s):  
Sotiris Kyriakou ◽  
William Cheung ◽  
Theodora Mantso ◽  
Melina Mitsiogianni ◽  
Ioannis Anestopoulos ◽  
...  
Keyword(s):  
Malignant Melanoma ◽  
In Vitro Model ◽  
Ros Generation ◽  
Intrinsic Apoptosis ◽  
Membrane Depolarisation ◽  
Vitro Model ◽  
Induced Apoptosis ◽  
Related Proteins ◽  
Metabolomic Approach

SummaryMelanoma is an aggressive and highly metastatic type of skin cancer where the design of new therapies is of utmost importance for the clinical management of the disease. Thus, we have aimed to investigate the mode of action by which a novel methylated analogue of L-Mimosine (e.g., L-SK-4) exerts its therapeutic potency in an in vitro model of malignant melanoma. Cytotoxicity was assessed by the Alamar Blue assay, oxidative stress by commercially available kits, ROS generation, caspase 3/7 activation and mitochondrial membrane depolarisation by flow cytometry, expression of apoptosis-related proteins by western immunoblotting and profiling of lipid biosynthesis by a metabolomic approach. Overall, higher levels of ROS, sphingolipids and apoptosis were induced by L-SK-4 suggesting that the compound’s therapeutic potency is mediated through elevated ROS levels which promote the upregulation of sphingolipid (ceramide) biosynthesis thus leading to the activation of both extrinsic and intrinsic apoptosis, in an experimental model of malignant melanoma.

Gemcitabine-induced apoptosis in 5637 cell line: an in-vitro model for high-risk superficial bladder cancer

2007 ◽  
Vol 18 (2) ◽  
pp. 179-185 ◽  
Author(s):  
Paola Gazzaniga ◽  
Ida Silvestri ◽  
Angela Gradilone ◽  
Susanna Scarpa ◽  
Stefania Morrone ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
High Risk ◽  
Cell Line ◽  
In Vitro Model ◽  
Superficial Bladder Cancer ◽  
Vitro Model ◽  
Induced Apoptosis

scholarly journals Effects of Hypericum perforatum extract on oxaliplatin-induced neurotoxicity: in vitro evaluations

2017 ◽  
Vol 72 (5-6) ◽  
pp. 219-226 ◽  
Author(s):  
Lorenzo Cinci ◽  
Lorenzo Di Cesare Mannelli ◽  
Anna Maidecchi ◽  
Luisa Mattoli ◽  
Carla Ghelardini
Keyword(s):  
Therapeutic Strategy ◽  
In Vitro Model ◽  
Caspase 3 ◽  
Hypericum Perforatum ◽  
Common Side Effect ◽  
Rat Astrocytes ◽  
Vitro Model ◽  
Ht 29 ◽  
Novel Therapeutic

Abstract Hypericum perforatum L. has been used for centuries as a natural remedy for the treatment of many disorders. Neuropathic pain is a common side effect of oxaliplatin-based chemotherapy and often the cause of therapy discontinuation. Thanks to its anti-inflammatory and analgesic effects, the use of H. perforatum may be a novel therapeutic strategy for neuropathy. The aim of this paper was to evaluate the effect of H. perforatum hydrophilic extract on an in vitro model of oxaliplatin-induced neurotoxicity. The antioxidant potential of extract was first evaluated in cell-free models by the thiobarbituric acid-reactive substances assay and nitro blue tetrazolium oxidation test; the ability of H. perforatum extract to reduce oxaliplatin-induced caspase-3 activity in rat astrocytes and its potential interference with the cytotoxic effects of oxaliplatin in a colorectal cancer in vitro model (HT-29 cells) were also evaluated. The extract showed a significant antioxidant effect and was able to reduce caspase-3 activity in rat astrocytes. Of note, the extract alone exerted a cytotoxic effect in HT-29 cells and did not reduce the cytotoxicity of oxaliplatin in HT-29 cells. These data suggest that H. perforatum could be used as a novel therapeutic strategy for counteracting chemotherapy-induced neuropathy.

scholarly journals Development of a Novel Experimental In Vitro Model of Isothiocyanate-induced Apoptosis in Human Malignant Melanoma Cells

2016 ◽  
Vol 36 (12) ◽  
pp. 6303-6310 ◽  
Author(s):  
THEODORA MANTSO ◽  
ARISTEIDIS P SFAKIANOS ◽  
AITHNE ATKINSON ◽  
IOANNIS ANESTOPOULOS ◽  
MELINA MITSIOGIANNI ◽  
...  
Keyword(s):  
Malignant Melanoma ◽  
In Vitro Model ◽  
Melanoma Cells ◽  
Human Malignant Melanoma ◽  
Vitro Model ◽  
Induced Apoptosis

CircRNA 001418 Promoted Cell Growth and Metastasis of Bladder Carcinoma via EphA2 by miR-1297

2020 ◽  
Vol 14 (1) ◽  
pp. 68-78 ◽  
Author(s):  
Guorui Peng ◽  
Hongxue Meng ◽  
Hongxin Pan ◽  
Wentao Wang
Keyword(s):  
Cell Proliferation ◽  
Cell Growth ◽  
Cytochrome C ◽  
Protein Expression ◽  
Bladder Carcinoma ◽  
In Vitro Model ◽  
Caspase 3 ◽  
Activity Levels ◽  
Vitro Model

Background: Cancer is one of the major causes of human deaths at present. It is the leading cause of deaths in developed countries. Moreover, Circular RNAs (circRNAs) have been discovered to play important roles in tumor genesis and development and are abnormally expressed in bladder cancer . Objective: The present study aims to investigate the anti-cancer effects of circ 001418 on bladder carcinoma and its possible mechanism. Methods: Quantitative PCR (qPCR) and gene chip were used to measure the circ 001418 expression. Cell proliferation and transfer, apoptosis and caspase-8 and caspase-3 activity levels were measured using MTT, Transwell assay, Flow cytometry. Caspase-3 and 9 activity levels, EphA2, cytochrome c and FADD protein expression were detected using Western blotting. Results: The expression of circ 001418 was increased in patients with bladder carcinoma. Over-expression of circ 001418 promoted cell proliferation and transfer, and reduced apoptosis in vitro model of bladder carcinoma. Down-regulation of Circ 001418 inhibited cell proliferation and transfer, and induced apoptosis in vitro model of bladder carcinoma. Meanwhile, Overexpression of circ 001418 induced EphA2 and cytochrome c protein expression, suppressed FADD protein expression in vitro model of bladder carcinoma by suppression of miR-1297. MiR-1297 reduced the pro-cancer effect of circ 001418 on apoptosis of bladder carcinoma. Conclusion: Results showed thatcircRNA 001418 promoted cell growth and metastasis of bladder carcinoma via EphA2 by miR-1297.

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