• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 2003년도 2003 Annual Meeting, BioExhibition and International Symposium
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• pp.129-131
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• 2003

Recombinant bioluminescent bacteria and cultured human cells were applied for toxicogenomic analysis of environmentally hazardous chemicals. Recombinant bioluminescent biosensing cells were used to detect and classify the toxicity caused by various chemicals. Classification of toxicity was realized based upon the chemicals' mode of action using DNA-, oxidative-, protein, and membrane-damage sensitive strains. As well, a simple double-layered cell culture system using Caco-2 cells and Hep G2 cells, which mimic the metabolic processes occurring in humans, such as adsorption through the small intestine and biotransformationin both the small intestine and the liver, was developed to investigate the toxicity of hazardous materials to humans. For a more in-depth analysis, a DNA microarray was used to study the transcriptional responses of Caco-2 and Hep G2 cells to benzo〔a〕pyrene.

• 대한한의학방제학회지
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• 제30권3호
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• pp.137-143
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• 2022

Objectives : Brain-Liver axis is an important target of the chronic human diseases. Hepatic steatosis is one of the most famous disorders in the chronic diseases. This study investigated the moderating effect of beneficial herbs on the fat accumulation, which is mediated by the LXR alpha-SREBP-1c signaling pathway. Methods : In order to confirm the SREBP-1c inhibitory effect, we performed immonoblotting ananlysis using HepG2 cells and Huh 7 cells treated by T0901317, the ligand of LXRα. Results : Forsythiae suspensa water extract (FSE) was not cytotoxicity in cell lines. FSE inhibited SREBP-1c protein expression in HepG2 and Huh7 cells induced by T0901317. In addition, FSE increased the phosphorylation of LKB1, which is associated with LXR-related pathway in HepG2 and Huh 7 cells. Conclusions : These results showed that FSE activated LKB1 to suppress SREBP-1c, which protects the cells against oxidative stress.

• Fisheries and Aquatic Sciences
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• 제15권1호
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• pp.21-27
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• 2012

Ground seawater in the east area of the volcanic Jeju Island contains abundant minerals. We investigated the metabolic activity of electrodialyzed, desalted ground seawater (EDSW) from Jeju in both cultured cells and animals. The addition of EDSW to the culture medium (up to 20%, v/v) reduced the leakage of lactate dehydrogenase and increased MTT activity in CHO-IR cells. EDSW (10%) promoted insulin-induced glucose consumption in L6 muscle cells as well as the activities of the liver ethanol-metabolizing enzymes, alcohol dehydrogenase and aldehyde dehydrogenase. Moreover, EDSW suppressed palmitate-induced intracellular fat accumulation in human hepatoma $HepG_2$ cells. Activities of AMP-stimulated protein kinase and acetyl CoA carboxylase, enzymes that modulate fat metabolism, were altered by EDSW in $HepG_2$ cells toward the suppression of intracellular lipid accumulation. EDSW also suppressed hepatic fat accumulation induced by a high-fat diet in mice. Taken together, EDSW showed beneficial metabolic effects, including the enhancement of ethanol metabolism and insulin-induced glucose consumption, and the suppression of intrahepatic fat accumulation.

• 대한한의학회지
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• 제29권4호
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• pp.171-179
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• 2008

Objective: Bufonis Venenum is the traditional Korean medicine Chan Su, which is obtained from the skin and parotid venom gland of the toads. It has been used for myocardial diseases, inflammation diseases, pain relief, cancer and others. The main components of BV are cinobufotoxin, cinobufalin, bufalin and others. Of these, bufalin, the major active ingredient of BV, has been reported to induce apoptosis and to possess anti-tumor effects. There was no report of anti-tumor screening of BV on hepatic cancer and which signaling pathway can be involved. In order to examine the effect of BV on hepatic cancer and the related signaling pathway with BV-induced apoptosis, human Hep G2 cells were used. Methods: Analysis of apoptosis was confirmed by MTT assay. BV decreased cell viability in a dose and duration dependent manner. To observe which signaling molecules will be activated by BV, phosphorylation of MAPK (p38, ERK, JNK), caspase 8 and caspase 9 were examined by Western blot analysis. Results: The phosphorylation levels of p38 started to increase at 5 min after addition of 5 ${\mu}g$/ml of BV and sustained to increase until 48 hours. The phosphorylation levels of other MAPK (ERK and JNK), caspase 8 and caspase 9 increased in a time-dependent manner. These imply that BV may activate different signaling pathways, MAPK, caspase 8 and caspase 9. These results propose that BV may induce apoptosis on Hep G2 cells through the activation of MAPK, caspase 8 and caspase 9.

• 대한본초학회지
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• 제22권3호
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• pp.27-37
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• 2007

Objectives: Hepatocellular carcinoma is the most common primary malignant tumor of the liver worldwide. In-Jin-Ho-Tang(IJHT) has been used as a traditional Chinese herbal medicine since ancient time. and today it is widely applied as a medication for jaundice which is associated with inflammation in liver. In this study, I investigated whether methanol extract of IJHT induced HepG2 cancer cell death. Methods: Cytotoxic activity of IJHT on HepG2 cells was using XTT assay. Apoptosis induction by Ros A in HCT116 cells was verified by the induction of cleavage of poly ADP-ribose polymerase (PARP). and activation of caspase-3, -8 and -9. The release of cytochrome c from mitochondria to cytosol. the level of Bcl-2 and Bax and the expression of p53 and p21 were examined by western blotting analysis. Furthermore, MAPKs activation was analyzed by western blotting analysis. Results: IJHT induced apoptosis in HepG2 cells. And treatment of IJHT resulted in the release of cytochrome c into cytosol, decreased anti-apoptotic Bcl-2, and increased pri-apoptotic Bax expression. IJHT markedly inactivated extracellular signal-regulated kinase (ERK1/2), and activated p38 mitogen-activated protein (MAP) kinase. Sodium orthovanadate (SOV), a phosphatase inhibitor, to reverse IJHT-induced ERK1/2 inactivation and SB203580, a specific p38 MAP Kinase inhibitor efficiently blocked apoptosis of HepG2. Thus, IJHT induces apoptosis in HepG2 cells via MAP kinase modulation. Conclusion: These results indicated that IJHT has some potential for use as an anti-cancer agent.

• Asian Pacific Journal of Cancer Prevention
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• 제14권11호
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• pp.6363-6367
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• 2013

Atractylis lancea (Thunb.) DC. (AL), an important medicinal herb in Asia, has been shown to have anti-tumor effects on cancer cells, but the involved mechanisms are poorly understood. This study focused on potential effects and molecular mechanisms of AL on the proliferation of the Hep-G2 liver cancer cell line in vitro. Cell viability was assessed by MTT test in Hep-G2 cells incubated with an ethanol extract of AL. Then, the effects of AL on apoptosis and cell cycle progression were determined by flow cytometry. Telomeric repeat amplification protocol (TRAP) assays was performed to investigate telomerase activity. The mRNA and protein expression of human telomerase reverse transcriptase (hTERT) and c-myc were determined by real-time RT-PCR and Western blotting. Our results show that AL effectively inhibits proliferation in Hep-G2 cells in a concentrationand time-dependent manner. When Hep-G2 cells were treated with AL after 48h,the $IC_{50}$ was about 72.1 ${\mu}g/mL$. Apoptosis was induced by AL via arresting the cells in the G1 phase. Furthermore, AL effectively reduced telomerase activity through inhibition of mRNA and protein expression of hTERT and c-myc. Hence, these data demonstrate that AL exerts anti-proliferative effects in Hep-G2 cells via down-regulation of the c-myc/hTERT/telomerase pathway.

• 생명과학회지
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• 제21권11호
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• pp.1625-1630
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• 2011

CoPP는 다양한 세포에서 HO-1의 유전자 발현과 활성을 증가시키는 강력한 유도제로 알려져 있다. HO-1는 세포 및 조직의 손상을 보호한다는 연구가 활발히 진행되고 있으나, 그 작용 기전에 대해서는 아직 잘 모르고 있다. 본 논문에서는 porphyrin 계열의 CoPP의 자극에 의해 유도되는 HO-1 유전자 발현에서 NADPH oxidase의 활성이 미치는 영향을 인간 간암세포주 HepG2에서 조사하였다. 배양 중인 HepG2 세포에서 CoPP는 HO-1의 발현을 농도의존적으로 증가시키는 것을 확인하였다. NADPH oxidase 저해제로 잘 알려져 있는 DPI를 전처리한 후 CoPP로 자극한 세포에서는 HO-1의 발현이 강력하게 억제되는 것으로 나타났다. DPI의 이런 억제 효과가 HO-1의 전사 조절인자 Nrf2의 활성에도 영향을 줄 수 있기 때문에 DPI를 전처리 한 후 CoPP 자극에 의한 Nrf2의 핵으로의 이동을 분석하였다. 그 결과, DPI는 CoPP에 의해 유도되는 Nrf2의 핵으로의 이동과 세포 내 존재하는 양을 감소시키는 것을 확인하였다. 다른 HO-1 발현 유도제로 알려져 있는 hemin에 의한 자극의 경우에도 DPI는 HepG2 세포의 HO-1의 발현을 억제하는 효과를 나타내었다. 그리고, $p47^{phox}$에 대한 siRNA를 사용하여 효과적으로 $p47^{phox}$ 유전자 발현을 knockdown 시켜서 NADPH oxidase의 활성을 억제시키는 방법을 사용하였다. 그 결과, $p47^{phox}$ silencing한 세포에 CoPP를 처리한 경우는 control siRNA를 처리한 세포와 비교할 때 HO-1 발현이 현저히 감소됨을 관찰할 수 있었다. 마지막으로, 세포 내 ROS 생성을 억제하는 GSHmee가 처리된 세포에서는 CoPP나 hemin이 Nrf2의 활성을 증가시키지 못하였고, 그 결과 HO-1의 발현을 유도하지 못하는 것을 알 수 있었는데, 이는 ROS가 CoPP나 hemin에 의한 HO-1 유전자 발현 과정에 중요한 역할을 한다는 것을 의미한다. 이를 종합해 볼 때, 인간 간암세포주 HepG2에서 CoPP나 hemin의 자극에 의한 HO-1 유전자의 발현에는 NADPH oxidase의 활성이 요구된다는 것을 알 수 있고, 그 활성은 세포 내 ROS를 생성시키는 것으로 역할을 한다고 여겨진다.

• Molecular & Cellular Toxicology
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• 제4권1호
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• pp.22-30
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• 2008

Malachite Green (MG), a toxic chemical used as a dye, topical antiseptic and antifungal agent for fish, is highly soluble in water, cytotoxic to various mammalian cells and also acts as a liver tumor promoter. In view of its industrial importance and possible exposure to human beings, MG possesses a potential environmental health hazard. So, we performed with HepG2, a human hepatocellular carcinoma cell line, to identify the differentially expressed genes (DEGs) related to toxicity of MG. And we compared gene expression between control and MG treatment to identify genes that are specifically or predominantly expressed by employing annealing control primer (ACP)-based $GeneFishing^{TM}$ method. The cytotoxicity $(IC_{20})$ of MG was determined above the $0.867{\mu}M$ in HepG2 cell for 48 h treatment. And the DEGs of MG were identified that 5 out of 6 DEGs were upregulated and 1 out of 6 DEGs was down-regulated by MG. Also, MG induced late apoptosis and necrosis in a dose dependent in flow cytometric analysis. Through further investigation, we will identify more meaningful and useful DEGs on MG, and then can get the information on mechanism and pathway associated with toxicity of MG.

• Asian Pacific Journal of Cancer Prevention
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• 제15권18호
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• pp.7785-7792
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• 2014

Background: Valproic acid (VPA) is a potent anticancer and antiangiogenic agent. However, design and synthesis of chemical derivatives with improved antiangiogenic and anticancer activities are still necessary. In this study a library of novel derivatives of VPA was synthesized and tested. Methods: A human liver cancer cell line (HepG2) and a human normal embryonic kidney cell line (HEK 293) were exposed to various concentrations of VPA derivatives for 24 hours and cell viability was checked by MTT colorimetric assay. Anti-angiogenic properties were evaluated in transgenic zebrafish embryos. Results: N-valproylglycine derivatives suppressed survival almost 70% (p value 0.001) in HepG2 cells but only 10-12% in HEK 293 cells (p value 0.133). They also suppressed angiogenic blood vessel formation by 80% when used between $2-20{\mu}M$ in zebrafish embryos. Valproic acid hydrazides showed moderate level of anticancer activity by affecting 30-50% (p value 0.001) of cell viability in HepG2 cells and 8-10% in HEK293 cells (p value 0.034). Conclusion: The majority of compounds in this study showed potent and stronger antiangiogenic and anticancer activity than VPA. They proved selectively toxic to cancer cells and safer for normal cells. Moreover, these compounds inhibited developmental angiogenesis in zebrafish embryos. Based on the fact that liver is a highly vascularized organ, in case of liver carcinoma these compounds have the potential to target the pathological angiogenesis and could be an effective strategy to treat hepatocellular carcinoma.

• BMB Reports
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• 제42권6호
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• pp.344-349
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• 2009

Angiogenesis is crucial for solid tumor growth. By secreting angiogenic factors, tumor cells induce angiogenesis. However, targeting these angiogenic factors for cancer therapy is not always successful, suggesting that other factors may be involved in tumor angiogenesis. This work shows that 25 protein spots were differentially expressed by two-dimensional gel electrophoretic analysis when HepG2 cells induced endothelial cell differentiation to tube in vitro, and most of them were upregulated. Twenty-one proteins were identified with MALDITOF-MS, and the other four were identified by LTQ-MS/MS. Keratins were identified as one class of these upregulated proteins. Further study indicated that the expression of keratin 17 in cultured endothelial cells is likely microenvironment regulated, because its expression can be induced by HepG2 cells and bFGF as well as serum in culture media. Increased expression of keratins in endothelial cells, such as keratin 17, may contribute to the angiogenesis induced by HepG2 cells.