• Biomolecules & Therapeutics
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• v.26 no.4
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• pp.399-408
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• 2018

In this study, we examined the molecular and functional characterization of choline uptake in the human esophageal cancer cells. In addition, we examined the influence of various drugs on the transport of [$^3H$]choline, and explored the possible correlation between the inhibition of choline uptake and apoptotic cell death. We found that both choline transporter-like protein 1 (CTL1) and CTL2 mRNAs and proteins were highly expressed in esophageal cancer cell lines (KYSE series). CTL1 and CTL2 were located in the plasma membrane and mitochondria, respectively. Choline uptake was saturable and mediated by a single transport system, which is both $Na^+$-independent and pH-dependent. Choline uptake and cell viability were inhibited by various cationic drugs. Furthermore, a correlation analysis of the potencies of 47 drugs for the inhibition of choline uptake and cell viability showed a strong correlation. Choline uptake inhibitors and choline deficiency each inhibited cell viability and increased caspase-3/7 activity. We conclude that extracellular choline is mainly transported via a CTL1. The functional inhibition of CTL1 by cationic drugs could promote apoptotic cell death. Furthermore, CTL2 may be involved in choline uptake in mitochondria, which is the rate-limiting step in S-adenosylmethionine (SAM) synthesis and DNA methylation. Identification of this CTL1- and CTL2-mediated choline transport system provides a potential new target for esophageal cancer therapy.

• Journal of Microbiology and Biotechnology
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• v.19 no.8
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• pp.803-809
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• 2009

Recent evidence has revealed the occurrence of an apoptotic phenotype in Candida albicans that is inducible with environmental stresses such as acetic acid, hydrogen peroxide, and amphotericin B. In the present study, we found that the Chinese herbal medicine Baicalein (BE), which was one of the skullcapflavones, can induce apoptosis in C. albicans. The apoptotic effects of BE were detected by flow cytometry using Annexin V-FITC and DAPI, and it was confirmed by transmission electron microscopy analysis. After exposure to 4 ${\mu}g$/ml BE for 12 h, about 10% of C. albicans cells were apoptotic. Both the increasing intracellular levels of reactive oxygen species (ROS) and upregulation of some redox-related genes (CAP1, SOD2, TRR1) were observed. Furthermore, we compared the survivals of CAP1 deleted, wild-type, and overexpressed strains and found that Cap1p attenuated BE-initiated cell death, which was coherent with a higher mRNA level of the CAP1 gene. In addition, the mitochondrial membrane potential of C. albicans cells changed significantly (p<0.001) upon BE treatment compared with control. Taken together, our results indicated that BE treatment induced apoptosis in C. albicans cells, and the apoptosis was associated with the breakdown of mitochondrial membrane potential.

• The Korean Journal of Physiology and Pharmacology
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• v.11 no.2
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• pp.45-55
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• 2007

We elucidated the effects of various components of ischemic medium on the outcome of simulated ischemia-reperfusion injury. Hypoxia for up to 12 hours induced neither apoptotic bodies nor LDH release. However, reoxygenation after 6 or 12 hours of hypoxia resulted in a marked LDH release along with morphological changes compatible with oncotic cell death. H9c2 cells were then subjected to 6 hours of simulated ischemia by exposing them to modified hypoxic glucose-free Krebs-Henseleit buffer. Lowered pH (pH 6.4) of simulated-ischemic buffer resulted in the generation of apoptotic bodies during ischemia, with no concomitant LDH release. The degree of reperfusion-induced LDH release was not affected by the pH of ischemic buffer. Removal of sodium bicarbonate from the simulated ischemic buffer markedly increased cellular damages during both the simulated ischemia and reperfusion. Addition of lactate to the simulated ischemic buffer increased apoptotic cell death during the simulated ischemia. Most importantly, concomitant acidosis and high lactate concentration in ischemic buffer augmented the reperfusion-induced oncotic cell death. These results confirmed the influences of acidosis, bicarbonate deprivation and lactate on the progression and outcome of the simulated ischemia-reperfusion, and also demonstrated that concomitant acidosis and high lactate concentration in simulated ischemic buffer contribute to the development of reperfusion injury.

• Journal of Pharmacopuncture
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• v.12 no.3
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• pp.49-59
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• 2009

Background : Anticancer effects of herbal medicine have been reported in various types of cancer, but the systematic approaches to explain molecular mechanism(s) are not established yet. Objective : The purpose of this study is to investigate the apoptotic cell death by Egg White combined Chalcanthite in NCI-H460 human lung cancer cells. Methods : Inhibitory effects were estimated by the MTT-assay. Cancer cells were stained with DAPI and showed condensed and fragmented nuclei. The expression of cleaved caspase-3, bcl-2, and bax was detected by western blotting. To establish a basis of understanding for anti-cancer mechanism, whole proteins have been obtained from NCI-H460 harvested at 24 hrs after the treatment of Egg White combined Chalcanthite, protein expression has been profiled by 2DE-based proteomic approach. Results : NCI-H460 human lung cancer cells were treated by three samples of IS3, IS4 and IS5. IS4 inhibited most effectively the growth of NCI-H460 human lung cancer cells. The expression of cleaved caspase-3 increased in IS4 in a concentration-dependent manner. Various changes of the protein expression have been monitored, and most frequent dysregulation was found in Vimentin, Lamin-A/C. Conclusion : Egg White combined-Chacanthite inhibited the growth of NCI-H460 human lung cancer cells by inducing the apoptotic cell death via caspase-3 activation. Based upon the present findings, the further study will focus on monitoring various cancer survival factors after artificial regulation of the proteins identified, and it would be the basis for the understanding of the Chacabthite anticancer effect(s) at the molecular level.

• Journal of Nutrition and Health
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• v.39 no.4
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• pp.357-365
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• 2006

DHA, one of w-3 fatty acids, modulates cell growth or death though the changes of apoptotic signaling in human endothelial ECV304 cells. We investigated the effects of DHA on the changes of apoptotic signaling in human vascular endothelial ECV304 cells using lipid peroxidation (LPO) metabolites. LPO could be originated by dietary polyunsaturated fatty acids such as linoleic acid(LA), arachidonic acid(AA) and docosahexaenoic acid (DHA). DHA caused cell death of ECV304 cells compared to LA, AA or control as evidenced by changes in cell morphology and MTT assay. LPO levels was significantly elevated by 10 fold in DHA-treated ECV 304 cells and caspase-3 activity was increased by DHA corresponding to increasing incubation times compared to control. One of reasons of the cell death in DHA-treated ECV304 cells could be expected that caspase activity, marker for mitochondrial damages, might be triggered by the increasing LPO levels. Our results strongly indicated that DHA induced LPO production has an important role on apoptotic signaling pathway in ECV304 cells. LPO production in endothelial cells which was metabolized by oxidation of dietary PUFA, might be one of risk factors in the initial progression of atherosclerosis.

• Development and Reproduction
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• v.5 no.2
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• pp.131-136
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• 2001

The developmental profiles of rpr, grim, dcp-1, diapl, diap2 transcripts, which were involved in programmed cell death, were analyzed using competitive RT-PCR in whole animals during Drosophila development. The fluctuation patterns of transcript levels of the apoptotic initiators(rpr and grim) were similar to those of the ecdysone titer in Drosophila life cycle. The transcript of dcp-1, which is considered as effector caspase, was expressed strong1y at early embryo and female adult stages. However, the transcript levels of anti-apoptotic factors diap1 and diap2, showed the reverse pattern comparing with those of apoptotic factors(rpr and grim). Also, the transcript levels of rpr, diap2 and dcp-1 were quantified in the salivary glands and wing discs dissected from the wandering late third instar larva. The transcript levels of rpr and diap2 were changed reversely each other in both tissues from wandering stage to puparium formation. These results suggest that the expressions of cell death related genes are regulated by the ecdysone signals during normal development.

• Journal of Physiology & Pathology in Korean Medicine
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• v.25 no.1
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• pp.71-77
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• 2011

Gastric ulcer has multifactorial etiology, and the development of ulcer is known to be caused by gastric acidity, pepsin secretion, gastric motility and gastric mucosal blood flow. The ulcer results from the tissue necrosis and apoptotic cell death triggered by mucosal ischemia, free radical formation and cessation of nutrient delivery. The gastric mucosa is usually exposed to a wide range of aggressive insults, and has developed efficient mechanisms to repair tissue injury. The apoptotic process of gastric mucosa is triggered by the induction of such proapoptotic gene expression, such as BAX. The Bcl-2 family of proteins plays a pivotal role in the regulation of apoptosis. The maintenance of gastric mucosa integrity depends upon the ratio between cell proliferation and cell death. Stress-inducing factors may affect Bcl-2/BAX ratio and thus the rate of apoptosis through modulation of the expression of both proteins depends upon the experimental model. In addition to the regulation of apoptosis, new vessels have to be generated in order to ensure an adequate supply of oxygen and nutrients to the healing gastric mucosa. This events are regulated by several factors. Among them, such polypeptide growth factors, such as vascular endothelial growth factor (VEGF) regulates essential cell functions involved in tissue healing including cell proliferation and differentiation. The purpose of this study was carried to investigate whether Rhei Rhizoma administration might protect apoptotic cell death and promote angiogenesis in gastric mucosa. Sprague-Dawley rats were randomly divided into 4 groups; normal, saline, cimetidine and Rhei Rhizoma-treated group. The saline, cimetidine and Rhei Rhizoma extracts were orally administrated to each group and gastric ulcer was induced by HCl-EtOH solution. After 1 hour, the stomachs were collected for histological observation and immunohistochemistry. In results, Rhei Rhizoma proves to promote to heal wound in gastric ulcer in conclusion and the significant changes of BAX, Bcl-2 and VEGF quantity in gastric mucosa were observed. These results suggest that Rhei Rhizoma extract may promote incision wound healing and has protective effects on gastric ulcer in rats.

• Journal of Chest Surgery
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• v.51 no.3
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• pp.187-194
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• 2018

Background: Death domain-associated protein (DAXX), originally identified as a pro-apoptotic protein, is now understood to be either a pro-apoptotic or an anti-apoptotic factor with a chromatin remodeler, depending on the cell type and context. This study evaluated DAXX expression and its clinical implications in squamous cell carcinoma of the esophagus. Methods: Paraffin-embedded tissues from 60 cases of esophageal squamous carcinoma were analyzed immunohistochemically. An immune reaction with more than 10% of tumor cells was interpreted as positive. Positive reactions were sorted into 2 groups: reactions in 11%-50% of tumor cells and reactions in more than 51% of tumor cells, and the correlations between expression and survival and clinical prognosticators were analyzed. Results: Forty-three of the 60 cases (71.7%) showed strong nuclear DAXX expression, among which 19 cases showed a positive reaction (31.7%) in 11%-50% of tumor cells, and 24 cases (40.0%) showed a positive reaction in more than 51% of tumor cells. A negative reaction was found in 17 cases (28.3%). These patterns of immunostaining were significantly associated with the N stage (p=0.005) and American Joint Committee on Cancer stage (p=0.001), but overall survival showed no significant difference. There were no correlations of DAXX expression with age, gender, or T stage. However, in stage IIB (p=0.046) and stage IV (p=0.014) disease, DAXX expression was significantly correlated with survival. Conclusion: This investigation found upregulation of DAXX in esophageal cancer, with a 71.7% expression rate. DAXX immunostaining could be used in clinical practice to predict aggressive tumors with lymph node metastasis in advanced-stage disease, especially in stages IIB and IV.

• Korean Journal of Pharmacognosy
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• v.36 no.2 s.141
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• pp.97-101
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• 2005

Saussureae Radix, the dried root of Saussurea lappa Clarke, has traditionally used for alleviating pain in abdominal distention and tenesmus, indigestion with anorexia, dysentery, nausea, and vomiting. Here we observed that methanol extracts of Saussurea Radix inhibited CDK2 activities in vitro. This inhibitory compound was isolated and identified as dehydrocostuslactone, one of the major constituents of Saussurea Radix. It is well known that dehydrocostuslactone induces apoptotic cell death. In this study, we also showed that dehydrocosruslactone inhibited cellular Rb phosphorylation and blocked cell growth at the concentration below $12\;{\mu}g/ml$ at which apoptotic cell death was not observed. Taken together, these results indicated that dehydrocostuslactone showed its anti-proliferative effects through the inhibition of CDK2 activity as well as the induction of apoptotic cell death.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.1
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• pp.101-109
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• 2004

Spatholobus suberectus belonging the family Leguminosae has been used for promoting blood circulation, removing blood stasis, tonifying the blood, relaxing tendons, stopping internal bleeding and eliminating dampness in oriental traditional medicine. This study investigates whether the water extracts of S. suberectus induce apoptotic cell death in Jurkat T-acute lymphoblastic leukemia (ALL) cells. Jurkat cells were increased inhibitions of cell viability in a concentration-dependent manner by S. suberectus, as measured by cell morphology. The capability of S. suberectus to induce apoptosis was associated with proteolytic cleavage of specific target protein such as poly (ADP­ribose)polymerase protein suggesting the possible involvement of caspases. The purpose of the present study is also to investigate the effect of S. suberectus on cell cycle progression. G1 checkpoin related gene products tested (cyclin D1, cyclin dependent kinase 4, retinoblastoma, E2Fl) were decreased in their protein levels in a dose-dependent manners after treatment of the extract. These results indicate that the increase of apoptotic cell death by S. suberectus may be due to the inhibition of cell cycle progression in wild type p53-lacking Jurkat cells.