• Korean Journal of Plant Resources
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• v.27 no.3
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• pp.215-222
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• 2014

This study was conducted to isolate a compound with anticancer properties from the roots of Peucedanum japonicum Thunb. (Umbelliferae), and to evaluate the efficacy of that compound's anticancer activity. The $CHCl_3$ layer was purified via repeated column chromatography and recrystallization. The two compounds isolated from $CHCl_3$ layer were identified via NMR spectroscopic analysis as (10E) 1,10-heptadecadiene-4,6-diyne-3,8,9-triol (Comp. I) and anomalin (Comp. II). (10E) 1,10-heptadecadiene-4,6-diyne-3,8,9-triol was the first report from the roots of P. japonicum. MTT assays were conducted to evaluate the in vitro cytotoxic activities of Compounds I and II against the following human cancer cell lines: HeLa, HepG2, SNU-16, and AGS. Comp. I evidenced the most profound cytotoxic activity against HepG2 cells ($IC_{50}=6.04{\mu}g/mL$), and Comp. II exhibited the most profound cytotoxic activity against SNU-16 cells ($IC_{50}=18.24{\mu}g/mL$) among the human cancer cell lines tested in this study. However, no significant cell death was observed in the CCD-25Lu human normal lung fibroblast cells. Quantitative analysis using UPLC (Ultra Performance Liquid Chromatography) showed that the roots of P. japonicum contained 0.015 (Comp. I) and 1.69 mg/g (Comp. II) of these compounds.

• Journal of Physiology & Pathology in Korean Medicine
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• v.27 no.6
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• pp.764-770
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• 2013

Stevia rebaudiana is a traditional herb used as a sweetener in Brazil and Paraguay as well as Korea and China. This study investigated the efficacy of Stevia rebaudiana methanol extract (SRE) to protect cells against the mitochondrial dysfunction and apoptosis in hepatocyte. To determine the effects of SRE on oxidative stress, we used the human derived hepatocyte cell line, HepG2 cell. Treatment of arachidonic acid (AA)+iron in HepG2 cells synergistically amplified cytotoxicity, as indicated by the excess reactive oxygen species (ROS) and mitochondrial permeability transition by fluorescence activated cell sorter (FACS) and immunoblot analysis. Treatment with SRE protected hepatocytes from AA+iron-induced cellular toxicity, as shown by alterations in the protein levels related with cell viability such as procaspase-3. SRE also prevented the mitochondrial dysfunction induced by AA+iron, and showed anti-oxidant effects as inhibition of $H_2O_2$ production and GSH depletion. Moreover, we measured the effects of SRE on AMP-activated protein kinase (AMPK), a key regulator in determining cell survival or death. Acetyl-CoA Carboxylase (ACC), a direct downstream target of AMPK. SRE increased phosphorylation of ACC, and prevented the inhibition of ACC phosphorylation by AA+iron. These results indicated that SRE has the ability to protect cells against AA+iron-induced $H_2O_2$ production and mitochondrial impairment, which may be mediated with AMPK-ACC pathway.

• Journal of Life Science
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• v.25 no.9
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• pp.1051-1058
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• 2015

Citrus mutant fruits were induced by irradiation of citrus budsticks with 120 Gy of cobalt (⁶⁰CO) gamma irradiation. The citrus mutant inhibited the growth and induced apoptosis in various human cancer cells, including A549, HepG2, HCT116, MCF-7, and Hela. The results of a trypan blue exclusion assay showed that citrus mutant fruits exhibited excellent antiproliferation activity in various human cancer cells and low cytotoxicity in normal 16HBE140- and CHANG cells. In addition, the cell death induced by the citrus mutant fruits was associated with an increased population of cells in sub-G1 phase, and it caused DNA fragmentation in human lung adenocarcinoma A549 and hepatocellular carcinoma HepG2 cells. It also up-regulated the amount of cellular nitric oxide (NO^•) produced as a result of nitric oxide synthase (NOS) activation and suppressed the inhibitor of apoptosis protein (IAP) family in A549 and HepG2 cells. These findings indicate that the citrus mutant fruits activates the NO^•-mediated apoptotic pathway in A549 and HepG2 cells. It may merit further investigation as a potential chemotherapeutic and chemopreventive agent for the treatment of various types of cancer cells. The results provide important major new insights into the mechanisms of the anticancer activity of citrus mutant fruits.

• Biomedical Science Letters
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• v.11 no.4
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• pp.487-492
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• 2005

The baculovirus/insect cell expression system is of great value in the study of structure-function relationships in mammalian glucose-transport proteins by site-directed mutagenesis and for the large-scale production of these proteins for mechanistic and biochemical studies. Spodoptera frugiperda Clone 21 (Sf2l) cells grow well on TC-100 medium that contains $0.1\%$ D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transporters. However, very little is known about the properties of the endogenous sugar transporter(s) in Sf2l cells, although a saturable transport system for hexose uptake has been previously revealed in the Sf cells. In order to further examine the substrate and inhibitor recognition properties of the Sf2l cell transporter, the ability of hexoses to inhibit 2-deoxy-D-glucose (2dGlc) transport was investigated by measuring inhibition constants $(K_i)$. The $K_i's$ for reversible inhibitors were determined from plots of uptake versus inhibitor concentration. Transport was effectively inhibited by D-mannose and D-glucose. Of the hexoses tested, L-glucose had the least effect on 2dGlc transport in the Sf2l cells, indicating that the transport is stereoselective. Unlike the human HepG2 type glucose transport system, D-mannose had a somewhat greater affinity for the Sf2l cell transporter than D-glucose, implying that the hydroxyl group at the C-2 position is not necessary for strong binding. However, epimerization at the C-4 position of D-glucose (D-galactose) resulted in a dramatic decrease in affinity of the hexose for the Sf2l cell transporter. Such a lowering of affinity might be the result of the involvement of the C-4 hydroxyl in hydrogen bonding. It is therefore suggested that Sf2l cells were found to contain an endogenous sugar transport activity that in several aspects resembles the human HepG2 type glucose transporter, although the insect and human transporters do differ in their affinity for cytochalasin B.

• The Korean Journal of Mycology
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• v.31 no.3
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• pp.155-160
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• 2003

Neutral salt soluble [0.9% NaCl (Fr. NaCl)], hot water soluble (Fr. HW) and methanol soluble (Fr. MeOH) materials were extracted from Paecilomyces sinclairii. In vitro cytotoxicity tests. Fr. HW was not cytotoxic against MH3T3, Sarcoma 180 and HepG2 cancer cell lines at the concentration of $0{\sim}2,000\;{\mu}g/ml$, while HT-29 cell line was cytotoxic at the concentration of $100\;{\mu}g/ml$. Fr. NaCl and Fr. MeOH were slightly cytotoxic to the cell lines. Intraperitoneal injection with Fr. NaCl and Fr. MeOH exhibited antitumor activity with life prolongation effect of 32.3% in mice inoculated with Sarcoma 180. Fr. NaCl improved proliferation of spleen cells and the immunopotentiation activity of B lymphocyte by increasing spleen cell numbers and the alkaline phosphatase activity by $2.4{\sim}2.6\;and\;2.7{\sim}3.9$ folds, respectively. Fr. NaCl generated $89\;{\mu}M$ of nitric oxide (NO) when cultured with RAW 264.7, a mouse macrophage cell line, at the concentration of $50\;{\mu}g/ml$, while iipopolysaccharide, a positive control, produced $79\;{\mu}M$. The Fr. NaCl showed the hightest antitumor effect and activation of B lymphocytes and macrophage. From these results, antitumor effect of P. sinclaitii was likely due to immunopotentiation activity.

• BMB Reports
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• v.29 no.2
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• pp.175-179
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• 1996

Viral surface proteins are known to play an essential role in attachment of the virus particle to the host cell membrane. In case of the hepatitis B virus (HBV) several reports have described potential receptors on the target cell side, but no definite receptor protein has been isolated yet. As for the viral side, it has been suggested that the preS region of the envelope protein, especially the preS1 region, is involved in binding of HBV to the host cell. In this study, preS1 region was recombinantly expressed in the form of a maltose binding protein (MBP) fusion protein and used to identify and visualize the expression of putative HBV receptor(s) on the host cell. Using laser scanned confocal microscopy and by FACS analysis, MBP-preS1 proteins were shown to bind to the human hepatoma cell line HepG2 in a receptor-ligand specific manner. The binding kinetic of MBP-preS1 to its cellular receptor was shown to be temperature and time dependent. In cells permeabilized with Triton X-100 and treated with the fusion protein, a specific staining of the nuclear membrane could be observed. To determine the precise location of the receptor binding site within the preS1 region, several short overlapping peptides from this region were synthesized and used in a competition assay. In this way the receptor binding epitope in preS1 was revealed to be amino acid residues 27 to 51, which is in agreement with previous reports. These results confirm the significance of the preS1 region in virus attachment in general, and suggest an internalization pathway mediated by direct attachment of the viral particle to the target cell membrane.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.255.1-255.1
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• 2002

We investigated the anti-proliferative effects of lxeris sonchifolia H. (godulbaegi) root extracts. luteolin(3'. 4', 5. 7-Q-glucoside and 3'. 4', 5, 7-tetrahydoxyflavone) and apigenin (3', 4'. 5. 7-O-gluconic acid) on HepG2 (p53 wild type) cells. Hep3B (p53 null) cells, and Chang liver cells. In MTT assay 3', 4'.5. 7-tetrahydoxyflavone showed the most efficient anti-proliferative effects on these three cell lines. However, there was no significant anti-proliferative effect on Chang liver cell line in MTT results. (omitted)

• Journal of Microbiology and Biotechnology
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• v.20 no.4
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• pp.821-827
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• 2010

Gene delivery that provides targeted delivery of therapeutic genes to the cells of a lesion enhances therapeutic efficacy and reduces toxic side effects. This process is especially important in cancer therapy when it is advantageous to avoid unwanted damage to healthy normal cells. Incorporating cancer-specific ligands that recognize receptors overexpressed on cancer cells can increase selective binding and uptake and, as a result, increase targeted transgene expression. In this study, we investigated whether a peptide capable of homing to hepatocellular carcinoma (HCC) could facilitate targeted gene delivery by cationic liposomes. This homing peptide (HBP) exhibited selective binding to a human hepatocarcinoma cell line, HepG2, at a concentration ranging from 5 to 5,000 nM. When conjugated to a cationic liposome, HBP substantially increased cellular internalization of plasmid DNA to increase the transgene expression in HepG2 cells. In addition, there was no significant enhancement in gene transfer detected for other human cell lines tested, including THLE-3, AD293, and MCF-7 cells. Therefore, we demonstrate that HBP provides targeted gene delivery to HCC by cationic liposomes.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.8
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• pp.3651-3657
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• 2014

Hepatocellular carcinoma (HCC) has a relatively higher incidence in many countries of Asia. Globally, HCC has a high fatality rate and short survival. Epirubicin, a doxorubicin analogue, may be administered alone or in combination with other agents to treat primary liver cancer and metastatic diseases. However, the toxic effects of epirubicin to normal tissues and cells have been one of the major obstacles to successful cancer chemotherapy. Here, we investigated the effects of epirubicin in combination with kappa-selenocarrageenan on mice with H22 implanted tumors and HepG-2 cell proliferation, immune organ index, morphology, cell cycle and related protein expressions in vivo and in vitro with sequential drug exposure. The inhibitory rate of tumor growth in vivo was calculated. Drug sensitivity was measured by MTT assay, and the King's principle was used to evaluate the interaction of drug combination. Morphological changes were observed by fluorescent microscopy. Cell cycle changes were analyzed by flow cytometry. Expression of cyclin A, Cdc25A and Cdk2 were detected by Western blotting. In vivo results demonstrated that the inhibitory rate of EPI combined with KSC was higher than that of KSC or EPI alone, and the Q value indicated an additive effect. In addition, KSC could significantly raise the thymus and spleen indices of mice with H22 implanted tumors. In the drug sensitivity assay in vitro, exposure to KSC and EPI simultaneously was more effective than exposure sequentially in HepG-2 cells, while exposure to KSC prior to EPI was more effective than exposure to EPI prior to KSC. Q values showed an additive effect in the simultaneous group and antagonistic effects in the sequential groups. Morphological analysis showed similar results to the drug sensitivity assay. Cell cycle analysis revealed that exposure to KSC or EPI alone arrested the cells in S phase in HepG-2 cells, exposure to KSC and EPI simultaneously caused accumulation in the S phase, an effect caused by either KSC or EPI. Expression of cyclin A, Cdc25A and Cdk2 protein was down-regulated following exposure to KSC and EPI alone or in combination, exposure to KSC and EPI simultaneously resulting in the lowest values. Taken together, our findings suggest that KSC in combination with EPI might have potential as a new therapeutic regimen against HCC.

• Food Science and Preservation
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• v.10 no.3
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• pp.394-400
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• 2003

This study was earned out to investigate the antimutagenic and cytotoxic effects of Eleutherococcus senticosus Maxim fruits ethanol extract on Salmonella typhimurium TA98, TA100 and cancer cell lines using Ames test and SRB assay, respectively. They were extracted with ethanol and then fractionated with hexane, chloroform, ethyl acetate, butanol and water to get active fractions. In the Ames test, most of the extracts had strong antimutagenic effects against the mutagenesis induced by MNNG, 4NQO, B(${\alpha}$)P and Trp-P-l. The ethanol extract (200 $\mu\textrm{g}$/plate) of Eleutherococcus senticosus fruits showed 87.2％ inhibitory effect on the mutagenesis induced by MNNG against TA100. And also, The suppression ratio against B(${\alpha}$)P and Trp-P-l in the TA100 showed 96.1％ and 95.5％, respectively. In the cytotoxic effects against human cancer cell lines (A549, AGS, MCF-7, Hep3B), the value of inhibition were mostly above 60％ for each fraction (1 mg/mL). Hexane fraction (1 mg/mL) against showed the strongest cytotoxic effects of 92.7％ compared to those of other fraction and butano fraetion against Hep3B was relatively high growth inhibitory effect of 82％.