• Asian Pacific Journal of Cancer Prevention
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• v.16 no.5
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• pp.1833-1837
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• 2015

An aristolactam-type alkaloid, isolated from Orophea enterocarpa, is enterocarpam-III (10-amino-2,3,4,6-tetramethoxyphenanthrene-1-carboxylic acid lactam). It is cytotoxic to various human and murine cancer cell lines; however, the molecular mechanisms remain unclear. The aims of this study were to investigate cytotoxic effects on and mechanism (s) of human cancer cell death in human hepatocellular carcinoma HepG2 and human invasive breast cancer MDA-MB-231 cells compared to normal murine fibroblast NIH3T3 cells. Cell viability was determined by MTT assay to determine $IC_{10}$, $IC_{20}$ and $IC_{50}$ levels, reactive oxygen species (ROS) production with 2',7'-dichlorohydrofluorescein diacetate and the caspase-3, -8 and -9 activities using specific chromogenic (p-nitroaniline) tetrapeptide substrates, viz., DEVD-NA, IETD-NA and LEHD-NA and employing a microplate reader. Mitochondrial transmembrane potential (MTP) was measured by staining with 3, 3'-dihexyloxacarbocyanine iodide ($DiOC_6$) and using flow cytometry. The compound was cytotoxic to HepG2 and MDA-MB-231 cells with the $IC_{50}$ levels of $26.0{\pm}4.45$ and $51.3{\pm}2.05{\mu}M$, respectively. For murine normal fibroblast NIH3T3 cells, the $IC_{50}$ concentration was $81.3{\pm}10.1{\mu}M$. ROS production was reduced in a dose-response manner in HepG2 cells. The caspase-9 and -3 activities increased in a concentration-dependent manner, whereas caspase-8 activity did not alter, indicating the intrinsic pathway activation. Enterocarpam-III decreased the mitochondrial transmembrane potential (MTP) dose-dependently in HepG2 cells, suggesting that the compound induced HepG2 cell apoptosis via the mitochondrial pathway. In conclusion, enterocarpam-III inhibited HepG2 and MDA-MB-231 cell proliferation and induced human HepG2 cells to undergo apoptosis via the intrinsic (mitochondrial) pathway and induction of caspase-9 activity.

• BMB Reports
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• v.30 no.4
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• pp.299-301
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• 1997

The asialoglycoprotein receptor (ASGPR) was the first described mammalian lectin that mediates the specific binding and internalization of galactose/N-acetylgalactosamine-terminating glycoproteins by hepatic parenchymal cells. H1 and H2 are known as essential subunits of the functional ASGPR. There were close similarities in ASGPR H2 subunits between cultured cell line HepG2 and normal human liver cells including identical sequences at both termini. It was therefore expected that there may be some similarities between the subunits from normal liver cells and fetal liver cells. The two subunits of human fetal liver ASGPR. designated FL-H1 and FL-H2. were cloned from cDNA library by peR and the sequences were compared with the known HI and H2 sequences of HepG2, and the H1 sequence of nornal human liver cells. The results showed that FL-H1 was identical to H1 of HepG2. Whereas FL-H2 contains a 15-bp miniexon, but missing 57-bp at the near upstream from the membrane-spanning domain compared to H2 of HepG2 and normal human liver cells indicating that FL-H2 resulted from a differential splicing compared to HepG2 and normal liver cells.

• The Journal of Korean Medicine
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• v.38 no.3
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• pp.43-58
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• 2017

Objectives: We performed this study to evaluate the antioxidative and hypolipidemic effect of Artemisia iwayomogi (韓茵蔯), Curcuma longa L. (鬱金) and Raphanus sativus L. (蘿?子) (ACR). Method: We enriched Artemisiae Capillaris, Curcumae Longae and Raphani Semen compound with alcohol. ACR extract is treated to HepG2 cell. Cell groups are devided into 3 groups: normal, control and ACR treated group. We measured polyphenol, flavonoids, DPPH and ABTS radical scavenging activity, ROS, glutathione, GSH peroxidase, GSH reductase, SOD, catalase, free fatty acid, lipid peroxidation and suppression of ACAT1 and HMG-CoA reductase expression on mRNA level. Results: 1. ACR contained polyphenol and flavonoids and increased GSH significantly in HepG2 cell. 2. ACR increased GPx, GR, and catalase activity significantly in HepG2 cell. 3. ACR increased DPPH and ABTS radical scavenging activity significantly in HepG2 cell and decreased ROS. 4. ACR decreased free fatty acid and MDA significantly in HepG2 cell. 5. ACR suppressed ACAT1 and HMG-CoA reductase expression on mRNA level in HepG2 cell. Conclusion: This study suggests that ACR has antioxidative and hypolipidemic effect and might be effective in prevention and treatment of dyslipidemia.

• Parasites, Hosts and Diseases
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• v.61 no.4
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• pp.388-396
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• 2023

Entamoeba histolytica is an enteric tissue-invasive protozoan parasite causing amoebic colitis and liver abscesses in humans. Amoebic contact with host cells activates intracellular signaling pathways that lead to host cell death via generation of caspase-3, calpain, Ca²⁺ elevation, and reactive oxygen species (ROS). We previously reported that various NADPH oxidases (NOXs) are responsible for ROS-dependent death of various host cells induced by amoeba. In the present study, we investigated the specific NOX isoform involved in ROS-dependent death of hepatocytes induced by amoebas. Co-incubation of hepatoma HepG2 cells with live amoebic trophozoites resulted in remarkably increased DNA fragmentation compared to cells incubated with medium alone. HepG2 cells that adhered to amoebic trophozoites showed strong dichlorodihydrofluorescein diacetate (DCF-DA) fluorescence, suggesting intracellular ROS accumulation within host cells stimulated by amoebic trophozoites. Pretreatment of HepG2 cells with the general NOX inhibitor DPI or NOX2-specific inhibitor GSK 2795039 reduced Entamoeba-induced ROS generation. Similarly, Entamoeba-induced LDH release from HepG2 cells was effectively inhibited by pretreatment with DPI or GSK 2795039. In NOX2-silenced HepG2 cells, Entamoeba-induced LDH release was also significantly inhibited compared with controls. Taken together, the results support an important role of NOX2-derived ROS in hepatocyte death induced by E. histolytica.

• Journal of Pharmacopuncture
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• v.15 no.4
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• pp.42-51
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• 2012

Objectives: This study used the basic principle of Oriental medicine, the sovereign, minister, assistant and courier principle (君臣佐使論) to investigate the effects of the component of ONGABO, which is composed of Ginseng Radix (Red Ginseng), Angelica Gigantis Radix, Schisandrae Fructus, Cuscuta Semen and Curcumae tuber on the viability of HepG2 cells. Methods: Single and mixed extracts of the component of ONGABO were prepared by lypohilizing powder of Red Ginseng (6-year root from Kanghwa), Angelica Gigantis Radix, Schisandrae Fructus, Cuscuta Semen, Curcumae Tuber (from Omniherb Co., Ltd., Korea) at the laboratory of herbal medicine in Woosuk University and were eluted after being macerated with 100% ethanol for three days. The cell viability of HepG2 was determined by using an absorptiometric analysis with PrestoBlue (Invitrogen) reagent after the plate had been incubated for 48 hours. All of the experiments were repeated three times to obtain the average value and standard deviation. The statistical analysis was done and the correlation factor was obtained by using Microsoft Office Excel 2007 and Origin 6.0 software. Results: Although Ginseng Radix (Red Ginseng) and Schisandrae Fructus did not enhance the viability of HepG2 cells, they were shown to provide protection of those cells. On the other hand, Angelica Gigantis Radix decreased the viability of HepG2 cells significantly, Cuscuta Semen and Curcumae Tuber had a small or no effect on the viability of HepG2 cells. Conclusions: In the sovereign, minister, assistant and courier principle (君臣佐使論), Ginseng Radix (Red Ginseng) corresponds to the sovereign component because it provides cell protection effects, Angelica Gigantis Radix corresponds to minister medicinal because it kills cells, Schisandrae Fructus corresponds to the assistant medicinal to help red ginseng having cell protect effects. Cuscuta Semen and Curcumae Tuber correspond to the courier medicinal having no effect in cell viability in HepG2. We hope this study provides motivation for advanced research on the sovereign, minister, assistant and courier principle.

• Journal of Physiology & Pathology in Korean Medicine
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• v.32 no.1
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• pp.13-23
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• 2018

This study was performed to investigate the antioxidant and antidyslipidemic effects of Artemisiae iwayomogii Herba, Curcumae longae Radix and Plantaginis Semen complex extract(ACP) on HepG2 cells. We measured total polyphenols, total flavonoids, radical scavenging activity, and ABTS radical scavenging activity of ACP to evaluate its antioxidant activity. HepG2 cells were treated with ACP. Then, we evaluated ROS production; intracellular GSH content; GPx, GR, SOD, and catalase activities; free fatty acids and MDA levels; and mRNA expression levels of ACAT1 and HMG-CoA reductase. Results: ACP contains polyphenols and flavonoids and increased the DPPH and ABTS radical scavenging activities in HepG2 cells in a dose dependent manner. Also, ACP significantly reduced ROS production in HepG2 cells compared to the control group and significantly increased the GSH content, and elevated the enzyme activities of GPx, GR, and catalase in HepG2 cells compared to the control group. In addition, ACP reduced the mRNA expression of ACAT1 and HMG-CoA reductase in HepG2 cells compared to that in the control group. Conclusion: These results suggest that ACP has an antioxidant effect and may suppress the expression of dyslipidemia - associated genes and thus may be useful for the improvement of dyslipidemia.

• Toxicological Research
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• v.22 no.4
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• pp.329-332
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• 2006

Ethanolic extract of Aloe vera (Aloe barbadensis Miller) was examined for the cellular toxicity on HepG2 human hepatocellular carcinoma cells. Treatment with Aloe vera extract resulted in DNA fragmentation but not LDH release, suggesting an apoptosis instead of necrosis. Aloe vera induced cytotoxicity was mediated by decrease in ATP levels, whereas GSH depletion, increase in intracellular $Ca^{2+}$, or activation of caspase-3/7 could not be observed with statistical significance. No activation of caspase-3/7 suggests the possibility of caspase-independent apoptosis. Taken together, our results show that Aloe vera extract induce HepG2 apoptosis by ATP depletion-related impairment of mitochondria, which is caspase-independent.

• The Korean Journal of Food And Nutrition
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• v.32 no.5
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• pp.565-570
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• 2019

Proteasome inhibitors can improve the efficiency of cancer treatments by inhibiting nuclear factor ${\kappa}B$($NF-{\kappa}B$) activation in cancer cells. Lentils are a type of beans of which consumption of such beans is increasing. The purpose of this study was to investigate the effects of lentils extract (LE) on the proteasomal activities, $NF-{\kappa}B$ activation, and cell cycle in HepG2 human liver cancer cells. LE treatments inhibited proteasomal activities at concentrations of 10, 50, and $100{\mu}g/mL$ respectively, and repressed $NF-{\kappa}B$ activation at concentrations of 1, 10, and $100{\mu}g/mL$ respectively, in HepG2 cells. LE treatments at concentrations of 1, 10, and $100{\mu}g/mL$ respectively, increased sub-G1 cell population in HepG2 cells, which may be the result of apoptosis. The results suggest that LE inhibited $NF-{\kappa}B$ activation partially with its proteasome inhibitory activities, and the increase of sub-G1 cell population was induced partially, by inhibition of $NF-{\kappa}B$ activation in HepG2 cells.

• The Journal of Korean Medicine
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• v.16 no.2 s.30
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• pp.414-432
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• 1995

In order to prove the antitumer effect of Bupleuri Radix(BR) and Artemisiae capillaris Herba(ACH) experimently, studies were done. The antitumer effect against hepatic cancer such as Hep G2, PLC & Hep 313, and also th synergastic action was evaulatcd in the combined treatment with anticancer drugs using chiefly for liver cancer, such as mitomycin(MMC), cisplatin(CPT) and 5-fluorouracil(5-FU). The results were obtained as follows: 1. IC50 against Hep G2, Hep 3B and PLC was 15.5ug/ml, 25.4ug/ml, 31.25ug/ml in Mitomycin (MMC), 92.5ug/ml, 50.2ug/ml, 62.5ug/ml in cisplatin(CPT) and 125ug/ml in 5-fluouracil(5- FU) respectively. 2. The antitumor effect was shown in the all concentrations of ACH, BR and below 55%-Cytotoxic effect against Hep G2 as compared with the date of control was shown in the concentration of $10^{-4}g/ml$ above of BR but not in ACH and also BR and ACHI revealed the synergistic effect with MMC. 3. The antitumor effect was shown in the concentration of $10^{-5}g/ml$ above of ACH, BR and below 55%-Cytotoxic effect against Hep 3B as compared with the data of control was shown in the concentration of $10^{-5}g/ml$ above of ACH but not in BH and also BR & ACH revealed the svnergistic effect with MMC. 4. The antitumor effect was shown in the all concentrations of ACH, BR and 55%-Cytotoxic effect against PLC as compared with the data of control was shown in the concentration of $10^{-5}g/ml$ above of ACH but not in BR and also ACH revealed the synergistic effect with MMC. From the above results it was concluded that Artemisiae capillaris had antitumor effect against PLC, Hep 3B, Bupleuri Radix against Hep G2 and also MMC showed the most synergistic effect in the anticancer drugs.

• YAKHAK HOEJI
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• v.56 no.2
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• pp.80-85
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• 2012

Previously, we have reported that arachidonic acid (AA) appears to be involved in the induction of apoptosis in HepG2 human hepatoblastoma cells. In this study we investigated the possible role of the NADPH oxidase, a membranebound enzyme generating reactive oxygen species (ROS), in the mechanism of AA-induced apoptosis in HepG2 cells. Apoptotic cell death induced by AA was significantly suppressed by various inhibitors of the NADPH oxidase, diphenylene iodonium (DPI), apocynin (Apo) and neopterine (NP). In addition, these inhibitors of the NADPH oxidase completely blunted the AA-induced ROS elevation. Next, we investigated the implication of metabolic pathway of AA in these AA actions. Both apoptosis and ROS production induced by AA were not significantly altered by treatment with indomethacin (Indo) or nordihydroguaiaretic acid (NDGA), selective inhibitors of cyclooxygenase (COX) and lipoxygenase (LOX), respectively, suggesting that AA metabolites produced by COX or LOX may not have an essential role in the AA-induced apoptosis and ROS generation. Collectively, these results suggest that the NADPH oxidase may be a key player in the mechanism of AA-induced apoptosis in HepG2 cells. These results further suggest that NADPH oxidase may be a good target for the management of human hepatomas.