• Title/Summary/Keyword: HepG2 human hepatoma cells

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Effects of Cholic Acid/CDCA and FGF-19 on the Protein Levels of the Endogenous Small Heterodimer Partner (SHP) in the Mouse Liver and HepG2 Cells (생쥐의 간과 HepG2 세포에 있어서 내인성 small heterodimer partner (SHP)의 단백질 수준에 미치는 cholic acid/CDCA 및 FGF-19의 효과)

  • Min, Gye-Sik
    • Journal of Life Science
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    • v.19 no.12
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    • pp.1731-1736
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    • 2009
  • Recent studies determined that a chronic western-style diet increased the endogenous small heterodimer partner (SHP) protein levels in mice. In experiments with cell cultures, chenodeoxy cholic acid (CDCA) treatment increased endogenous SHP protein levels and reduced the degradation rate of exogenously expressed flag-SHP levels in the human hepatoma cell line, HepG2 cells. In addition, bile acid-induced intestinal fibroblast growth factor-19 (FGF-19) increased the half-life of the exogenously expressed SHP when HepG2 cells were transfected with ad-flag-SHP. However, both the expression level and the degradation rate of the endogenous SHP in response to cholic acid and FGF-19 have not been well understood, either in mice or in cultured HepG2 cells. This study examined the effects of cholic acid treatment on the endogenous SHP protein levels in mice and the effects of FGF-19 on the degradation rate of the endogenous SHP protein in HepG2 cells. Mice fed 0.5% cholic acid in normal chow showed an increase in endogenous SHP protein levels during both 12 hr and 24 hr treatment periods as compared to control mice fed only normal chow. In cultured HepG2 cells, treatment with CDCA did not noticeably change the rate of degradation in the endogenous SHP protein from cells not treated with CDCA. Although consistent with the previous studies on the exogenous ad-flag-SHP protein, treatment with FGF-19 significantly decreased the degradation rate of the endogenous SHP protein when HepG2 cells were treated with cyclohexamide. These results suggest that both bile acids and FGF-19 increase the endogenous SHP protein levels in mouse liver and HepG2 cells.

MUTAGENICITY AND ANTIMUTAGENICITY OF CATECHIN AND TANNINS FROM THE BARK OF HAMAMELIS VIRGINIANA L. IN HUMAN Hep G2 CELLS

  • Sundermann, Volker-Mersch;Andreas Dauer;Evelyne Lhoste;Siegfried Knasmuller
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2001.10a
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    • pp.95-96
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    • 2001
  • The mutagenic and antimutagenic activities of catechin, hamamelitannin and two proanthocyanidin fractions prepared from the bark of Hamamelis virginia L. - a commonly used medicinal herb - were investigated in a human derived hepatoma (Hep G2) cell line using the single cell gel electrophoresis (SCGE, syn. Comet assay) for the detection of DNA-migration. The cells possess different phase I and phase II enzymes involved in the biotransformation of xenobiotics.(omitted)

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Establishment of a Hepatocellular Carcinoma Cell Line Expressing Dual Reporter Genes: Sodium Iodide Symporter (NIS) and Enhanced Green Fluorescence Protein (EGFP) (나트륨 옥소 공동수송체 유전자와 녹색 형광 유전자의 이중 리포터 유전자를 발현하는 간암세포주 확립)

  • Kwak, Won-Jung;Koo, Bon-Chul;Kwon, Mo-Sun;Lee, Yong-Jin;Lee, Hwa-Young;Yoo, Jeong-Soo;Kim, Te-Oan;Chun, Kwon-Soo;Cheon, Gi-Jeong;Lee, Sang-Woo;Ahn, Byeong-Cheol;Lee, Jae-Tae
    • Nuclear Medicine and Molecular Imaging
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    • v.41 no.3
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    • pp.226-233
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    • 2007
  • Purpose: Dual reporter gene imaging has several advantages for more sophisticated molecular imaging studies such as gene therapy monitoring. Herein, we have constructed hepatoma cell line expressing dual reporter genes of sodium iodide symporter (NIS) and enhanced green fluorescence protein (EGFP), and the functionalities of the genes were evaluated in vivo by nuclear and optical imaging. Materials and Methods: A pRetro-PN vector was constructed after separating NIS gene from pcDNA-NIS. RSV-EGFP-WPRE fragment separated from pLNRGW was cloned into pRetro-PN vector. The final vector expressing dual reporter genes was named pRetro-PNRGW. A human hepatoma (HepG2) cells were transfected by the retrovirus containing NIS and EGFP gene (HepG2-NE). Expression of NIS gene was confirmed by RT-PCR, radioiodine uptake and efflux studies. Expression of EGFP was confirmed by RT-PCR and fluorescence microscope. The HepG2 and HepG2-NE cells were implanted in shoulder and hindlimb of nude mice, then fluorescence image, gamma camera image and I-124 microPET image were undertaken. Results: The HepG2-NE cell was successfully constructed. RT-PCR showed NIS and EGFP mRNA expression. About 50% of cells showed fluorescence. The iodine uptake of NIS-expressed cells was about 9 times higher than control. In efflux study, $T_{1/2}$ of HepG2-NE cells was 9 min. HepG2-NE xenograft showed high signal-to-background fluorescent spots and higher iodine-uptake compared to those of HepG2 xenograft. Conclusion: A hepatoma cell line expressing NIS and EGFP dual reporter genes was successfully constructed and could be used as a potential either by therapeutic gene or imaging reporter gene.

Vitexin, an HIF-1α Inhibitor, Has Anti-metastatic Potential in PC12 Cells

  • Choi, Hwa Jung;Eun, Jae Soon;Kim, Bang Geul;Kim, Sun Yeou;Jeon, Hoon;Soh, Yunjo
    • Molecules and Cells
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    • v.22 no.3
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    • pp.291-299
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    • 2006
  • Vitexin, a natural flavonoid compound identified as apigenin-8-C-${\beta}$-D-glucopyranoside, has been reported to exhibit antioxidative and anti-inflammatory properties. In this study, we investigated its effect on hypoxiainducible factor-$1{\alpha}$ (HIF-$1{\alpha}$) in rat pheochromacytoma (PC12), human osteosarcoma (HOS) and human hepatoma (HepG2) cells. Vitexin inhibited HIF-$1{\alpha}$ in PC12 cells, but not in HOS or HepG2 cells. In addition, it diminished the mRNA levels of hypoxia-inducible genes such as vascular endothelial growth factor (VEGF), smad3, aldolase A, enolase 1, and collagen type III in the PC12 cells. We found that vitexin inhibited the migration of PC12 cells as well as their invasion rates, and it also inhibited tube formation by human umbilical vein endothelium cells (HUVECs). Interestingly, vitexin inhibited the hypoxia-induced activation of c-jun N-terminal kinase (JNK), but not of extracellular-signal regulated protein kinase (ERK), implying that it acts in part via the JNK pathway. Overall, these results suggest the potential use of vitexin as a treatment for diseases such as cancer.

Inhibition of SMP30 Gene Expression Influences the Biological Characteristics of Human Hep G2 Cells

  • Zhang, Sheng-Chang;Liang, Ming-Kang;Huang, Guang-Lin;Jiang, Kui;Zhou, Su-Fang;Zhao, Shuang
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.3
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    • pp.1193-1196
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    • 2014
  • Senescence marker protein 30 (SMP30), a hepatocellular carcinoma (HCe) associated antigen had been identified by our research group. To study its mechanisms of regulation and associations with the occurrence and development of HCe, we inhibited expression by RNAi technique, and observed effects on the biological characteristics of Hep G2 cells. In cell viability assays, cell growth in the experimental group (with siRNA transfection) was elevated. In Transwell invasion assays, compared with blank and control groups, numbers of invading cells in the experimental group were significantly increased, whereas in apoptosis assays, the percentage apoptosis demonstrated no differences, but after UV irradiation, that in the experimental group was higher than the other two groups. In a word, SMP30 can inhibit the proliferation and invasion of human hepatoma cells and thus can be regarded as a cancer suppressive factor.

Metabolic Activity of Desalted Ground Seawater of Jeju in Rat Muscle and Human Liver Cells

  • Kim, Bo-Youn;Lee, Young-Ki;Park, Deok-Bae
    • Fisheries and Aquatic Sciences
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    • v.15 no.1
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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.

Effect of Ginseng Components on Content of Cholesterol and Activity of Acyl CoA.Cholesterol Acyltransferase in Hep G2 Cells Cultured in Cholesterol Rich Medium (고콜레스테를 조건으로 배양한 Hep G2세포의 콜레스테를 함량변동과 Acyl CoA : Cholesterol Acyltransferase의 활성에 미치는 인삼성분의 영향)

  • Park, Song-Chul;Noh, Yun-Hee;Koo, Ja-Hyun
    • Journal of Ginseng Research
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    • v.19 no.3
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    • pp.212-218
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    • 1995
  • A human hepatoma cell line, hep G2, was used to investigate the mechanism of serum cholesterol reduction by ginseng total saponin, ginsenoside-$Rb_1$, - $Rb_2$, and non-saponin fraction (ether extraction). Hep G2 cells were incubated in 10 $\mu\textrm{g}$/ml of cholesterol containing serum free-RPMl1640 medium with various concentration of ginseng components. The amounts of cholesterol in Hep G2 cells were decreased to maximum 51% in total saponin or two ginsenoside-treated groups while there was 137% increase in cholesterol level of control group as compared with that of normal group. Nonsaponin groups did not show the same effect. In order to elucidate the observed changes in the amount of cholesterol, the activity of amyl CoA : cholesterol acyltransferase (ACAT) in groups showing remarkable reduction in cholesterol amount, i.e., total saponin 10-6%, ginsenoside-$Rb_1$ $10^{-4}$%, ginsenoside-$Rb_2$, $10^{-4}$%, and non-saponin fraction $10^{-4}$%, was assayed using [1-$^{-14}C$%]oleic acid as enzyme substrate. The activity of ACAT was increased in all groups tested as compared with that of control group except for non-saponin group cultured in water soluble cholesterol containing medium. The serum cholesterol lowering effects of ginseng components can partially be attributed to the increased hepatocellular ACAT activity.

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Cooperative stimulation of cisplatin-mediated apoptosis by hepatitis B virus X Protein and hepatitis C virus core Protein (B형 간염 바이러스 X 단백질과 C형 간염 바이러스의 코어 단백질에 의한 cisplatin-매개성 세포 예정사의 협조적 촉진)

  • Kwun, Hyun-Jin;Jang, Kyung-Lib
    • Journal of Life Science
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    • v.17 no.6 s.86
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    • pp.766-771
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    • 2007
  • The co-infection with hepatitis B virus (HBV) and hepatitis C Virus (HCV) is associated with a more severe liver disease and increased frequency in the development of hepatocellular carcinoma com-pared to those with single infection. Here, we demonstrated that HBV X protein (HBx) and HCV Core cooperatively up-regulated the level of p53 in human hepatoma HepG2 cells. The elevated p53 subsequently stimulated the expression of proapoptotic Bax whereas it repressed the expression of antiapoptotic Bcl2. These effects, however, were not observed in p53-negative Hep3B cells. Consistently to their cooperative regulation of apoptotic effectors, HBx and HCV Core additively stimulated cisplatin-mediated apoptotic cell death of HepG2 but not of Hep3B cells. These results may help to explain the development of a more severe liver disease in patients co-infection with HBV and HCV as well as some contradictory results on the roles of HBx and Core in apoptosis.

In Vitro Free Radical Scavenging and Hepatoprotective Activities of Taraxacum mongolicum (포공영의 자유라디칼 소거 및 간세포 보호활성)

  • Baek, Hum-Young
    • Korean Journal of Pharmacognosy
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    • v.34 no.4 s.135
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    • pp.324-326
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    • 2003
  • The methanol (MeOH) extract and ist fractions of Taraxacum mongolicum (Compositae) were examined for their scavenging effects on 1,1-diphenyl-2- phenylhydrazyl (DPPH) and superoxide radicals, and hepatoprotective effects on tacrine-induced cytotoxicity in human hepatoma cell line, Hep G2 cells. Both methylene chloride $(CH_2Cl_2)$ and butanol (n-BuOH) soluble fractions of the MeOH extract showed the free radicals scavenging and hepatoprotective effects. From these results, it is suggested that hepatoprotective effect of these fractions partly relies on their free radical scavenging activity.

The Effect of Artemisia Capillaris Herba on Ethanol-Induced Cytokines(TNF-${\alpha},IL-1{\alpha}$) Secretion in Hep G2 Cells (인진호가 Hep G2 세포에서 에탄올 매개성 Cytokine 분비에 미치는 영향)

  • Sim, Jung-Sub;Kim, Il-Hwan;Kim, Gang-San;Kagn, Byung-Ki;Choi, Su-Deock
    • The Journal of Internal Korean Medicine
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    • v.22 no.1
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    • pp.87-93
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    • 2001
  • A human hepatoma cell line, Hep G2 cells, is reliable for the study of alcohol-induced hepatotoxicity. The aim of this study is to determine the relationship between TNF-${\alpha}$, IL-$1{\alpha}$ production and EtOH-induced cytotoxicity on Hep G2 cells. The cells were incubated with EtOH in the presence of Artemisia Capillaris Herba(AC) for 24 hours and in the absence of AC for 48 hours. Cytoviability and cytokines release were analyzed by MTT assay and enzyme linked immunosorbent assay (ELISA), respectively. After 24 hours of EtOH exposure, the cytoviability had markedly decreased, and the release of cytokines had increased. The increased amount of cytokines contributed to EtOH-induced cytotoxicity. Anti-TNF-${\alpha}$ and IL-$1{\alpha}$ antibodies almost abolished it. Interestingly, EtOH-induced cytotoxicity and cytokines production were inhibited by AC. Moreover, when AC was used in combination with antibodies, there was a marked inhibition of EtOH-induced cytotoxicity. These results suggest that EtOH-induced cytotoxicity may regulate, by various factors, and AC may prevent the cytotoxicity through partial inhibition of the $TNF-{\alpha}$ and IL-$1{\alpha}$ secretion.

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