• BMB Reports
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• 제41권10호
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• pp.745-750
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• 2008

Selenium, an essential trace element possessing anti-carcinogenic properties, can induce apoptosis in cancer cells. We have previously shown that sodium selenite can induce apoptosis by activating the mitochondrial apoptosis pathway in NB4 cells. However, the detailed mechanism remains unclear. Presently, we demonstrate that p53 contributes to apoptosis by directing signaling at the mitochondria. Immunofluorescent and Western blot procedures revealed selenite-induced p53 translocation to mitochondria. Inhibition of p53 blocked accumulation of reactive oxygen species (ROS) and loss of mitochondrial membrane potential, suggesting that mitochondrial p53 acts as an upstream signal of ROS and activates the mitochondrial apoptosis pathway. Selenite also disrupted cellular calcium ion homeostasis in a ROS-dependent manner and increased mitochondrial calcium ion concentration. p38 kinase mediated phosphorylation and mitochondrial translocation of p53. Taken together, these results indicate that p53 involves selenite-induced NB4 cell apoptosis by translocation to mitochondria and activation mitochondrial apoptosis pathway in a transcription-independent manner.

• Reproductive and Developmental Biology
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• 제35권4호
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• pp.427-434
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• 2011

Mitochondria diseases have been reported to involve structural and functional defects of complex I-V. Especially, many of these diseases are known to be related to dysfunction of mitochondrial proton-translocating NADH-ubiquinone oxidoreductase (complex I). The dysfunction of mitochondria complex I is associated with neurodegenerative disorders, such as Parkinson's disease, Huntington's disease, and Leber's hereditary optic neuropathy (LHON). Mammalian mitochondrial proton-translocating NADH-quinone oxidoreductase (complex I) is largest and consists of at least 46 different subunits. In contrast, the NDI1 gene of Saccharomyces cerevisiae is a single subunit rotenone-insensitive NADH-quinone oxidoreductase that is located on the matrix side of the inner mitochondrial membrane. The Saccharomyces cerevisiae NDI1 gene using a recombinant adeno-associated virus vector (rAAV-NDI1) was successfully expressed in AML12 mouse liver hepatocytes and the NDI1-transduced cells were able to grow in media containing rotenone. In contrast, control cells that did not receive the NDI1 gene failed to survive. The expressed Ndi1 enzyme was recognized to be localized in mitochondria by confocal immunofluorescence microscopic analyses and immunoblotting. Using digitonin-permeabilized cells, it was shown that the NADH oxidase activity of the NDI1-transduced cells was not affected by rotenone which is inhibitor of complex I, but was inhibited by antimycin A. Furthermore, these results indicate that Ndi1 can be functionally expressed in the AML12 mouse liver hepatocytes. It is conceivable that the NDI1 gene is powerful tool for gene therapy of mitochondrial diseases caused by complex I deficiency. In the future, we will attempt to functionally express the NDI1 gene in mouse embryonic stem (mES) cell.

• 생명과학회지
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• 제10권4호
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• pp.340-346
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• 2000

This study was designed to investigate the effects of silk fibroin(Mw 500) powder (SFP) on oxygen radicals and the scavenger enzymes in brain membranes of rats. Spragu-Dawley(SD) male rats(160${\pm}$10g) were fed basic diet(control group), and experimental diets(SFP-2.5 and SFP-5.0 groups) added 2.5 and 5.0g/kg BW/day for 6 weeks. Hydroxyl radical($.$OH) levels resulted in a decreases(6.6% and 9.7%, 2.8% and 11.9%, respectively) in brain mitochondria and microsomes of SFP-2.5 and SFP-5.0 groups compared with control group, but were significantly decreased in these membrances of SFP-5.0 group only. Superoxide radical (O2) levels were a slightly decreased (2.0% and 9.1%, respectively) in brain cytosol of SFP-2.5 and SFP-5.0 groups compared with control group. Lipid peroxide(LPO) levels were significantly decreased (12.9% and 21.9%, 13.2% and 22.5%, respectively) in brain mitochondria and microsomes of SFP-2.5 and SFP-5.0 groups compared with control group. Oxidized protein (OP) levels were significantly decreased (16.7% and 15.7%, respectively) in brain microsomes of SFP-2.5 and SFP-5.0 group compared with control group, but significantly difference between in brain mitochondria of these two groups could not be obtained. Mn-SOD activities were remarkably increased (11.2% and 24.2%, respectively) in mitochodria of SFP-2.5 and SFP-5.0 groups. CuZn-SOD activities were effectively increased (7.7% and 19.6%, respectively) in brain cytosol of SFP-2.5 and SFP-5.0 groups, but significant difference between control and SFP-2.5 groups could be not obtained. GSHPx activities were considerably increased (5.3% and 11.7%, respectively) in brain cytosol of SFP-2.0 and SFP-5.0 groups compared with control group. There results suggest that anti-aging effect of silk fibroin may play an effective learning and memory role in a attenuating a oxidative stress and increasing a scavenger enzyme activity in brain membranes.

• The Korean Journal of Physiology and Pharmacology
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• 제21권6호
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• pp.599-607
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• 2017

Most normal cells express L-type amino acid transporter 2 (LAT2). However, L-type amino acid transporter 1 (LAT1) is highly expressed in many tumor cells and presumed to support their increased growth and proliferation. This study examined the effects of JPH203, a selective LAT1 inhibitor, on cell growth and its mechanism for cell death in Saos2 human osteosarcoma cells. FOB human osteoblastic cells and Saos2 cells expressed LAT1 and LAT2 together with their associating protein 4F2 heavy chain, but the expression of LAT2 in the Saos2 cells was especially weak. JPH203 and BCH, a non-selective L-type amino acid transporter inhibitor, potently inhibited L-leucine uptake in Saos2 cells. As expected, the intrinsic ability of JPH203 to inhibit L-leucine uptake was far more efficient than that of BCH in Saos2 cells. Likewise, JPH203 and BCH inhibited Saos2 cell growth with JPH203 being superior to BCH in this regard. Furthermore, JPH203 increased apoptosis rates and formed DNA ladder in Saos2 cells. Moreover, JPH203 activated the mitochondria-dependent apoptotic signaling pathway by upregulating pro-apoptotic factors, such as Bad, Bax, and Bak, and the active form of caspase-9, and downregulating anti-apoptotic factors, such as Bcl-2 and Bcl-xL. These results suggest that the inhibition of LAT1 activity via JPH203, which may act as a potential novel anti-cancer agent, leads to apoptosis mediated by the mitochondria-dependent intrinsic apoptotic signaling pathway by inducing the intracellular depletion of neutral amino acids essential for cell growth in Saos2 human osteosarcoma cells.

• 동의생리병리학회지
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• 제23권6호
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• pp.1241-1246
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• 2009

It has been suggested that Tae-Eum peoples are prone to obesity. Although extensive clinical observations have shown this tendency in Sasang Constitutional Medicine (SCM), no scientific hypothesis has been proposed to delineate its mechanism. According to SCM theory, Tae-Eum peoples have a hypoactive lung system and a hyperactive liver system. In this paper we propose a new hypothesis explaining the tendency of obesity in Tae-Eum people in the viewpoint of cell physiology. The hypoactive lung system might imply an attenuated 'respiration' at the cell/subcell level, namely mitochondrial oxygen consumption. Because a functional weakness in mitochondria energy metabolism indicates intrinsic hypo-activity in the consumption (or production) of metabolic energy, we deduced that the tendency can easily induce body weight gain via an increase in anabolism. This relation is also introduced in the graph of cellular metabolic power against body weight. To test this hypothesis, we analyzed the clinical data with 863 subjects. Statistical analysis of the data showed that Tae-Eum peoples had relatively a lower cellular metabolic power, and that the percentage of peoples with BMI>25 was significantly higher than that of the other constitutional types.

• BMB Reports
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• 제49권2호
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• pp.69-70
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• 2016

Mutations of orthodentricle homeobox 2 (OTX2) in human and mice often cause retinal dystrophy and nyctalopia, suggesting a role of OTX2 in mature retina, in addition to its functions in the development of the eye and retina. In support of this, the number of bipolar cells in Otx2^+/− post-natal mouse retina was found to be significantly lower than normal. Degeneration of the cells becomes greater as the mice age, leading to the loss of vision. Especially, the type-2 OFF-cone bipolar cells, which do not express Otx2 mRNA but carry Otx2 protein, are most sensitive to Otx2 haplodeficiency. Interestingly, this bipolar cell subpopulation imports Otx2 protein from photoreceptors to protect itself from glutamate excitotoxicity in the dark. Moreover, in the bipolar cells, the exogenous Otx2 relocates to the mitochondria to support mitochondrial ATP synthesis. This novel mitochondrial activity of exogenous Otx2 highlights the therapeutic potential of Otx2 protein transduction in retinal dystrophy.

• BMB Reports
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• 제38권6호
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• pp.709-716
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• 2005

Apoptosis and necrosis are distinguished by modality primarily. Here we show an apoptosis occurred instantly, induced by $300\;{\mu}M$ W-7 ((N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide hydrochloride), inhibitor of calmodulin), which demonstrated necrotic modality. As early as 30 min after W-7 addition, apoptotic (sub-diploid) peak could be detected by fluorescence-activated cell sorter (FACS), “DNA ladders” began to emerge also at this time point, activity of caspase-3 elevated obviously within this period. Absence of mitochondrial membrane potential (MMP) reduction and cytochrome c, AIF (apoptosis inducing factor) release, verified that this rapid apoptosis did not proceed through mitochondria pathway. Activation of caspase-12 and changes of other endoplasmic reticulum (ER) located proteins ascertained that ER pathway mediated this necrosis-like apoptosis. Our findings suggest that it is not credible to judge apoptosis by modality. Elucidation of ER pathway is helpful to comprehend the pathology of diseases associated with ER stress, and may offer a new approach to the therapy of cancer and neurodegenerative diseases.

• Animal cells and systems
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• 제13권1호
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• pp.49-57
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• 2009

For the study of the reproductive mechanism associated with the process of vitellogenesis, oocyte development and vitellogenesis during oogenesis in female Boleophthalmus pectinirostris were investigated by electron microscopic observations. The ovary consists of a pair of saccular structures with many ovarian lobules. In the early vitellogenic oocyte, the Golgi complex plays an important role leading to the formation of yolk vesicles containing carbohydrate yolks. At this time many pinocytotic vesicles containing yolk precursors are observed in the cytoplasm near the region of initial formation of the zona radiata. In the late vitellogenic oocytes, the multivesicular bodies, which are formed by modified mitochondria, are involved in the formation of the primary yolk granules. Precursors of yolk granules and multivesicular bodies develop to primary yolk globules with participation of pinocytotic vesicles. After primary yolk globules mix with each other, they develop into secondary and tertiary yolk globules. Based on these findings, vitellogenesis of B. pectinirostris occurs by way of the processes of endogenous autosynthesis and exogenous heterosynthesis. The process of autosynthesis involves the combined activity of the Golgi complex, mitochondria, and multivesicular bodies. However, the process of heterosynthesis involves pinocytotic incorporation of extraovarian precursors into the zona radiata of vitellogenic oocytes by way of the thecal cell layers and granulosa cells.

• BMB Reports
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• 제35권1호
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• pp.127-133
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• 2002

Nitrosative stress can prevent or induce apoptosis. It occurs via S-nitrosylation by the interaction of nitric oxide (NO) with the biological thiols of proteins. Cellular redox potential and non-heme iron content determine S-nitrosylation. Apoptotic cell death is inhibited by S-nitrosylation of the redox-sensitive thiol in the catalytic site of caspase family proteases, which play an essential role in the apoptotic signal cascade. Nitrosative stress can also promote apoptosis by the activation of mitochondrial apoptotic pathways, such as the release of cytochrome c, an apoptosis-inducing factor, and endonuclease G from mitochondria, as well as the suppression of NF-${\kappa}B$ activity. In this article we reviewed the mechanisms whereby S-nitrosylation and nitrosative stress regulate the apoptotic signal cascade.

• 대한핵의학회지
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• 제33권1호
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• pp.1-10
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• 1999

Cancer cells are known to show increased rates of glycolysis metabolism. Based on this, PET studies using F-18-fluorodeoxyglucose have been used for the detection of primary and metastatic tumors. To account for this increased glucose uptake, a variety of mechanisms has been proposed. Glucose influx across the cell membrane is mediated by a family of structurally related proteins known as glucose transporters (Gluts). Among 6 isoforms of Gluts, Glut-1 and/or Glut-3 have been reported to show increased expression in various tumors. Increased level of Glut mRNA transcription is supposed to be the basic mechanism of Glut overexpression at the protein level. Some oncogens such as src or ras intensely stimulate Glut-1 by means of increased Glut-1 mRNA levels. Hexokinase activity is another important factor in glucose uptake in cancer cells. Especially hexokinase type II is considered to be involved in glycolysis of cancer cells. Much of the hexokinase of tumor cells is bound to outer membrane of mitochondria by the porin, a hexokinase receptor. Through this interaction, hexokinase may gain preferred access to ATP synthesized via oxidative phosphorylation in the inner mitochondria compartment. Other biologic factors such as tumor blood flow, blood volume, hypoxia, and infiltrating cells in tumor tissue are involved. Relative hypoxia may activate the anaerobic glycotytic pathway. Surrounding macrophages and newly formed granulation tissue in tumor showed greater glucose uptake than did viable cancer cells. To expand the application of FDG PET in oncology, it is important for nuclear medicine physicians to understand the related mechanisms of glucose uptake in cancer tissue.