• Biomolecules & Therapeutics
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• v.30 no.2
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• pp.184-190
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• 2022

Targeting the cystine/glutamate exchange transporter, system x_c^-, is a promising anticancer strategy that induces ferroptosis, which is a distinct form of cell death mediated by iron-dependent lipid peroxidation. The concentration of ⳑ-cystine in culture medium is higher than the physiological level. This study was aimed to evaluate the effects of ⳑ-cystine concentration on the efficacy of ferroptosis inducers in hepatocellular carcinoma cells. This study showed that treatment with sulfasalazine or erastin, a system x_c^- inhibitor, decreased the viability of Huh6 and Huh7 cells in a dose-dependent manner, and the degree of growth inhibition was greater in medium containing a physiological ⳑ-cystine concentration of 83 µM than in commercial medium with a concentration of 200 µM ⳑ-cystine. However, RSL3, a glutathione peroxidase 4 inhibitor, decreased cell viability to a similar extent in media containing both ⳑ-cystine concentrations. Sulfasalazine and erastin significantly increased the percentages of propidium iodide-positive cells in media with 83 µM ⳑ-cystine, but not in media with 200 µM ⳑ-cystine. Sulfasalazine- or erastin-induced accumulation of lipid peroxidation as monitored by C11-BODIPY probe was higher in media with 83 µM ⳑ-cystine than in media with 200 µM ⳑ-cystine. In contrast, the changes in the percentages of propidium iodide-positive cells and lipid peroxidation by RSL3 were similar in both media. These results showed that sulfasalazine and erastin, but not RSL3, were efficacious under conditions of physiological ⳑ-cystine concentration, suggesting that medium conditions would be crucial for the design of a bioassay for system x_c^- inhibitors.

• Microbiology and Biotechnology Letters
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• v.44 no.4
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• pp.535-539
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• 2016

The prevalence of pathogenic bacteria such as Streptococcus parauberis (Sp), Streptococcus iniae (Si), and Edwardsiella tarda (Et) in flounder fish farms in Jeju Island and their management by gallium treatment was studied. Sp, Si, and Et were found to exhibit a low rate of cell growth and high biofilm formation. Hence, in the present study, cell growth and biofilm formation were measured spectrophotometrically 72 h after the addition of different concentrations of gallium (2, 4, or 8 mg/ml). In addition, cell death was measured by resazurin and propidium iodide staining assays. The results showed that bacterial cell death increased and biofilm formation decreased with an increasing concentration of gallium. Hence, the present study signifies that the use of gallium against bacterial pathogens could be useful for disease management in flounder farms.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.2
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• pp.358-364
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• 2006

To investigate the possible mechanism of Drynariae Rhizoma extracts as a candidate of anti-cancer drug, I examined the effects of Drynariae Rhizoma extracts on the apoptosis of U937 cell line. MTT assay, flow cytometric analysis, SDS-polyacrylamide gel electrophoresis, Western blot analysis, and RT-PCR were performed. Drynariae Rhizoma extracts treatment reduced the cell viablilty of U937 cells in a dose-dependent manner, which was associated with induction of apoptotic cell death. Drynariae Rhizoma extracts treatment also reduced the levels of Bcl-xL anti-apoptotic protein expression and increased the levels of caspase-3, p53, pro-apoptotic protein, in U937 cells. RT-PCR data revealed that the level of bcl-2, bcl-xL mRNA expressions decreased in a dose-dependent manner. These findings suggest that Drynariae Rhizoma extracts may have induction of apoptotic cell death via regulation of several growth regulatory gene products. The abbreviations used are: FBS, fetal bovine serum; PBS, phosphate buffered saline; PI, propidium iodide; OD, optical density; DiOC6, 3,3-dihexyloxa carbcyanine iodide; MTT, 3 [4-5-dimethylthiazol-2-yl] -2-diphenyltetrazolium bromide.

• Natural Product Sciences
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• v.10 no.2
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• pp.74-79
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• 2004

Dietary flavonoids are currently receiving considerable attention in developing novel cancer-preventive approaches because of their potential capacities to actively induce apoptosis of cancer cells. In our previous report, a flavonoid fraction, which consisted mainly of protocatechuic acid, fustin, fisetin, sulfuretin, and butein and named RCMF (RVS chloroform-methanol fraction), was prepared from a crude acetone extract of Rhus verniciflua Stokes (RVS) that is traditionally used as food additive and herbal medicine. In this study, we evaluated the effects of the RCMF on cell proliferation and apoptosis using SV40-transformed tumorigenic hepatocytes, BNL SV A.8. Tritium uptake assay showing the proliferative capacity of the cells was strongly suppressed in the presence of RCMF. This anti-proliferative effect was further confirmed through trypan blue exclusion. RCMF-mediated suppression of cell growth was verified to be apoptotic, based on the increase in DNA fragmentation, low fluorescence intensity in nuclei after propidium iodide staining, and the appearance of DNA laddering. Collectively, this study demonstrated that RCMF can be approached as a potential agent that is capable of significantly inhibiting cell growth of hepatic cancer cells.

• Biomolecules & Therapeutics
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• v.21 no.5
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• pp.358-363
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• 2013

In the present study, we investigated the effect of intracellular glutathione (GSH) depletion in heart-derived H9c2 cells and its mechanism. L-buthionine-S,R-sulfoximine (BSO) induced the depletion of cellular GSH, and BSO-induced reactive oxygen species (ROS) production was inhibited by glutathione monoethyl ester (GME). Additionally, GME inhibited BSO-induced caspase-3 activation, annexin V-positive cells, and annexin V-negative/propidium iodide (PI)-positive cells. Treatment with rottlerin completely blocked BSO-induced cell death and ROS generation. BSO-induced GSH depletion caused a translocation of PKC-${\delta}$ from the cytosol to the membrane fraction, which was inhibited by treatment with GME. From these results, it is suggested that BSO-induced depletion of cellular GSH causes an activation of PKC-${\delta}$ and, subsequently, generation of ROS, thereby inducing H9c2 cell death.

• Proceedings of the KSAR Conference
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• 2002.06a
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• pp.83-83
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• 2002

This study was to compare the characteristics of parthenogenetic mES (P-mES) cells and in vitro fertilization mES cells. Mouse oocytes were recovered from superovulated 4wks hybrid F1 (C57BL/6xCBA/N) female mice. The oocytes were treated with 7% ethanol for 5 min and 5 ㎍/㎖ cytochalasin-B for 4 h. For IVF, the oocytes were inseminated with epididymal sperm of hybrid Fl male mice (1×10/sup 6//㎖). IVF and parthenogenetic embryos were cultured in M16 medium for 4 days. Cell number count in blastocysts was carried out differential labelling using propidium iodide (red) and bisbenzimide(blue). (omitted)

• International Journal of Oral Biology
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• v.32 no.2
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• pp.45-49
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• 2007

In this study, the cytotoxicity of commonly used local anesthetics was evaluated on odontoblasts which are essential for pulpal homeostasis in vitro. Local anesthetics, such as articaine, bupivacaine, levobupivacaine, lidocaine, mepivacaine, prilocaine, and procaine, were tested on the odontoblast cell line, MDPC-23. The concentration-and time-dependent cytotoxic effects of local anesthetics on odontoblasts were measured by MTT assay. Among local anesthetics treated for 18 h, only bupivacaine significantly showed cell death in a concentration-($LC_{50}=1.2mM$) and time-dependent manner. To confirm cell death induced by bupivacaine, the observation of cell morphology and FACS using Annexin V and propidium iodide (PI) staining were performed. As a result of Annexin V and PI staining, as well as the morphological change, only bupivacaine induced apoptotic cell death on odontoblasts when compared with levobupivacaine and lidocaine. These results suggest that bupivacaine might affect normal pulpal integrity even after uneventful local anesthesia.

• Food Science and Biotechnology
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• v.16 no.4
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• pp.632-635
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• 2007

This study examined the neuroprotective effect of scopoletin from Angelica dahurica against oxygen and glucose deprivation-induced neurotoxicity in a rat organotypic hippocampal slice culture. Scopoletin reduced the propidium iodide (PI) uptake, which is an indication of impaired cell membrane integrity. In addition, it inhibited the loss of NeuN, which represents the viability of neuronal cells. The results suggests that scopoletin from A. dahurica protects neuronal cells from the damage caused by oxygen and glucose deprivation.

• Korean Journal of Clinical Pharmacy
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• v.9 no.2
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• pp.103-108
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• 1999

The anti-proliferating effects of some plants on hepatoma cell lines were studied by the 3-[4,5-dim-ethylthiazol-2yl]-2,5-diphenyl-tetrazolium bromide (MTT assay), to investigate the anticancer effect with some plants around here. As the result, we saw that the anti-prolferating effect to the plants. Among the plants, Equisetum arvense L. and Lactuca dentata Makino. var, flaviflora Makino of them relatively showed a good ant-proliferating effect. Capsicum annuum L. var. angulosum Mill (Leaf) was the best among them. We also examined morphological changes on the hepatoma cells in this process. In case of Capxicum annuum L. var. angulosum Mill, the tells become vague after 2 days, and then destroyed faster than others. We can fee also the condensated chromosome on the treated cells with Capxicum annuum L. var. angulosum Mill. And we also observed condensation through using a fluorescent microscope by PI staining, and observed DNA fragmentation.

• Journal of Ginseng Research
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• v.27 no.2
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• pp.52-55
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• 2003

The present experiment was perf'3rmed to investigate the protective effects of ginseng total saponin (GTS) and possible mechanisms on the hepatocytotoxicity induced by tert-butylhydroperoxide (t-BuOOH), 4-Bromo-calciumu ionophore A23187 (Br-A23187) and KCN. Hepatocytes were isolated by collagenase perfusion of livers from fasted male Sprague Dawley rats and cultured overnight. After various treatments in Krebs-Ringer-HEPES buffer at pH 7.4, cell viability was determined by propidium iodide using fluorocytometry. GTS (5-20 ${\mu}$M) inhibited cell killing induced by t-BuOOH, and KCN, dose-dependently. However, GTS did not inhibit Br-A23187-induced cell killing. These findings support that GTS could protect the hepatocytoxicity induced by some toxic chemicals. The mechanisms of these protective effects by GTS seem to be associated with antioxidant activity and increase of cellular ATP.