• Title/Summary/Keyword: human HepG2 cells

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Participation of SRE4, an URE1 Enhancer Core Sequence, in the Sterol-Mediated Transcriptional Upregulation of the Human Apolipoprotein E Gene

  • Min, Jung-Hwa;Paik, Young-Ki
    • BMB Reports
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    • v.31 no.6
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    • pp.565-571
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    • 1998
  • The expression of the endogenous human apolipoprotein(apo)E gene was significantly induced when HepG2 cells were treated with exogenous 25-hydroxy-cholesterol. This sterol-mediated apoE gene upregulation appears to require the participation of a positive element for the apoE gene transcription (PET) ( -169/ -140), a core sequence of upstream regulatory element (URE)1 enhancer of the human apoE gene. This PET was renamed as sterol regulatory element (SRE)4 based on its new role as a sensor for the level of intracellular sterol. Furthermore, a gel mobility shift analysis showed that binding activity of the SRE4 binding protein (BP) obtained from HepG2 cells was induced by sterol treatment, while that from either MCF7 or BT20 cells remained unchanged. Binding activity of SRE4BP was also induced in mouse macrophage cells, J774A.1, by sterol treatment, but it was drastically reduced when cells were subjected to treatment of AY-9944, a potent inhibitor for sterol synthesis. However, binding activity of Spl, which is a co-binding protein to the SRE4 region, remained the same in either condition, suggesting that SRE4BP (formally known as PETBP) may be mainly responsible for the sterol-mediated regulation of the apoE gene expression. Deletion analysis of the core binding site of SRE4BP by gel mobility shift assays showed that the minimal sequence of the SRE4BP binding appears to reside between -157 and -140, confirming the identity of SRE4 with the previously determined core sequence of URE1.

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Biostability and Drug Delivery Efficiency of γ-Fe2O3 Nano-particles by Cytotoxicity Evaluation (세포독성 평가를 통한 γ-Fe2O3 나노입자의 생체안정성 및 약물전달효율)

  • Lee, Kwon-Jai;An, Jeung-Hee;Shin, Jae-Soo;Kim, Dong-Hee;Yoo, Hwa-Seung;Cho, Chong-Kwan
    • Korean Journal of Materials Research
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    • v.20 no.3
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    • pp.132-136
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    • 2010
  • This study examined the biostability and drug delivery efficiency of g-$Fe_2O_3$ magnetic nanoparticles (GMNs) by cytotoxicity tests using various tumor cell lines and normal cell lines. The GMNs, approximately 20 nm in diameter, were prepared using a chemical coprecipitation technique, and coated with two surfactants to obtain a water-based product. The particle size of the GMNs loaded on hangamdan drugs (HGMNs) measured 20-50 nm in diameter. The characteristics of the particles were examined by X-ray diffraction (XRD), field emission scanning electron microscopy (FE-TEM) and Raman spectrometer. The Raman spectrum of the GMNs showed three broad bands at 274, 612 and $771\;cm^1$. A 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay showed that the GMNs were non-toxic against human brain cancer cells (SH-SY5Y, T98), human cervical cancer cells (Hela, Siha), human liver cancer cells (HepG2), breast cancer cells (MCF-7), colon cancer cells (CaCO2), human neural stem cells (F3), adult mencenchymal stem cells (B10), human kidney stem cells (HEK293 cell), human prostate cancer (Du 145, PC3) and normal human fibroblasts (HS 68) tested. However, HGMNs were cytotoxic at 69.99% against the DU145 prostate cancer cell, and at 34.37% in the Hela cell. These results indicate that the GMNs were biostable and the HGMNs served as effective drug delivery vehicles.

Cytotoxicity of Petroleum Ether Extract of Euonymus alatus (귀전우 Petroleum Ether 추출물의 세포독성)

  • Eun, Jae-Soon;Park, Sang-Ho;Kweon, Jin;Kim, Young-Ahn;Kang, Sung-Young;Oh, Chan-Ho;Jeon, Hoon
    • Korean Journal of Pharmacognosy
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    • v.26 no.3
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    • pp.253-258
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    • 1995
  • The purpose of this research was to investigate the effects of petroleum ether extract of Euonymus alatus (EAP) on the proliferation of human tumor cells. EAP inhibited the proliferation of HeLa, Hep G2, KHOS/NP and A431 cells. The cytotoxicity of doxorubicin on human tumor cells and Balb/c 3T3 cells were increased by the combination of EAP. EAP did not affect the proliferation of Balb/c 3T3 cells, mouse spleen cells and human lymphocytes. These results suggest that EAP has the cytotoxicity on human tumor cells without cytotoxicity on Balb/c 3T3 cells, mouse spleen cells and human lymphocytes, and increase the cytotoxicity of doxorubicin.

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Glycoantigen Biosyntheses of Human Hepatoma and Colon Cancer Cells are Dependent on Different N-Acetylglucosaminyltransferase-III and -V Activities

  • Kim, Cheorl-Ho
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.891-900
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    • 2004
  • UDP-N-Acetylglucosamine(GlcNAc):$\beta$1,4-D-mannoside$\beta$-l ,4N-acetylglucosaminyltransferase-III (GnT-III) and UDP-N-GlcNAc:$\alpha$-6-D-mannosid$\beta$-1,6N-acetylglucosaminyltransferase-V(GnT - V) activities were determined in human hepatoma cell lines and metastatic colon cancer cells, and their activities were compared with those of normal liver cells and fetal hepatocytes. GnT-III activities were higher than those of GnT-V in hepatic carcinoma cells. When the two enzyme activities were assayed in highly metastatic colon cancer cells, GnT - V activities were much higher than those of GnT-III. When GlcN, GlcN-biant-PA and UDP-GlcNAc were used as substrates, the enzymes displayed different kinetic properties between hepatic and colon cancer cells, depending on their metastatic potentials. Normal cells of two origins had characteristically very low levels of GnT-III and -V activities, whereas hepatoma and colon cancer cells contained high levels of activities. These data were supported by RT-PCR and Northern blot analyses, showing that the expression of GnT-III and -V mRNAs were increased in proportion to the enzymatic activities. The increased GnT-III, md -V activities were also correlated with increased glycosylation of the cellular glycoproteins in hepatoma and colon cancer cells, as examined by lectin blotting analysis by using wheat germ glutinin (WGA), erythroagglutinating phytohemagglutinin (E-PHA), leukoagglutinating phytohemagglutinin (L-PHA), and concanavalin A (Con A). Treatment with retinoic acid, a differentiation agent, resulted in decreases of both GnT-III and -V activities of HepG2 and HepG3 cells. In colon carcinoma cells, however, treatment with retinoic acid resulted in a reduction of GnT-V activity, but not with GnT-III activity. Although the mechanism underlying the induction of these mzymes is unclear, oligosaccharides in many glycoproteins have been observed of cancer cells.

Effect of Water Extract from Artemisiae Argi Folium on Hepatotoxicity Caused by Acetaminophen and Acetaldehyde (Acetaminophen과 Acetaldehyde로 유발된 간세포독성에 대한 애엽 물추출물의 영향)

  • Park, Wan-Su
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.5
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    • pp.1210-1214
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    • 2008
  • The purpose of this study is to investigate the effect of water extract from Artemisiae Argi Folium (WAAF) on hepatotoxicity caused by acetaminophen (AAP) and acetaldehyde which are regarded as hepatotoxin. Artemisiae Argi Folium was known to have the antibacterial, immune-enhancing, and anticoagulative properties. In Korean Medicine, Artemisiae Argi Folium is supposed to be related with 'liver meridian' according to traditional medical theory. AAP and acetaldehyde reduce the intracellular production of hydrogen peroxide ($H_2O_2$) and nitric oxide (NO) production of human hepatocyte HepG2. The intracellular production of hydrogen peroxide ($H_2O_2$) was measured by dihydrorhodamine 123 (DHR) assay. NO production was measured with Griess test. WAAF increased the production of $H_2O_2$ and NO reduced by AAP and acetaldehyde in HepG2 cells. Therefore, It could be suggested that WAAF has the hepatoprotective activity against AAP and acetaldehyde.

Generation and Characterization of Monoclonal Antibodies against Human Interferon-lambda1

  • Hong, Seung-Ho;Kim, Jung-Sik;Park, Sun
    • IMMUNE NETWORK
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    • v.8 no.1
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    • pp.7-12
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    • 2008
  • Background: Members belonging to the interferon-lambda (IFN-${\lambda}$) family exert protective action against viral infection; however, the mechanisms of their action have remained elusive. To study IFN-${\lambda}$ biology, such as endocytosis of IFN-${\lambda}$, we produced monoclonal antibodies (Abs) against human IFN-${\lambda}$ and examined their usefulness. Methods: We purified recombinant human IFN-${\lambda}$1 expressed in Escherichia coli by using affinity columns. Then, we generated hybridoma cells by fusing myeloma cells with splenocytes from IFN-${\lambda}$1-immunized mice. For evaluating the neutralizing activity of the monoclonal Abs against IFN-${\lambda}$1, we performed RT-PCR for the MxA transcript. In order to study the binding activity of IFN-${\lambda}$ and the monoclonal Ab complex on HepG2 cells, we labeled the monoclonal Ab with rhodamine and determined the fluorescence intensity. Results: Four hybridoma clones secreting Abs specific to IFN-${\lambda}$1 were generated and designated as HL1, HL2, HL3, and HL4. All the Abs reacted with IFN-${\lambda}$1 in the denatured form as well as in the native form. Abs produced by HL1, HL3, and HL4 did not neutralize the induction of the MxA gene by IFN-${\lambda}$1. We also demonstrated the binding of the HL1 monoclonal anbitody and IFN-${\lambda}$ complex on HepG2 cells. Conclusion: Monoclonal Abs against IFN-${\lambda}$1 were produced. These Abs can be used to study the cellular binding and internalization of IFN-${\lambda}$.

Antitumor Constituents from Anthriscus Sylvestris (L.) Hoffm

  • Chen, Hui;Jiang, He-Zhong;Li, Yong-Chao;Wei, Guo-Qing;Geng, Yun;Ma, Chao-Ying
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.6
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    • pp.2803-2807
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    • 2014
  • Bioassay-guided chemical investigation of the roots of Anthriscus sylvestris (L.) Hoffm. resulted in the isolation of nine compounds, whose structures were determined by spectroscopic methods. Compound 1 was isolated from this plant for the first time and compounds 3 and 9 were first found from this genus. Different polar fractions of A. sylvestris extract and compounds 1, 6-8 and 9 were evaluated for antitumor activities against HepG2 (human hepatocellular carcinoma), MG-63 (human osteosarcoma cells), B16 (melanoma cells) and HeLa (human cervical carcinoma cells) lines by the MTT method. The petroleum ether fraction of A. sylvestris extract exhibited excellent inhibitory activity with an $IC_{50}$ value of $18.3{\mu}g/ml$. Among the isolates from the petroleum ether fraction, compound 7 showed significant inhibition against the growth of the four tumor cells with $IC_{50}$ values ranging from $12.2-43.3{\mu}g/ml$.

The Effects of Anticarcinogenic Activity of Solanum tuberosum Peel Fractions (감자 껍질 분획성분의 항발암 효과)

  • 배송자
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.5
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    • pp.905-909
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    • 2002
  • This study was peformed to determine the anticarcinogenic activity of the Solanum tuberosum Peel (SP) on several microorganisms and human cancer cell lines. Among the various solvent fractions of SP, the ethylether Partition layer (SPMEE) showed the strongest antimicrobial activity, ethylacetate partition layer (SPMEA) and butanol partition layer (SPMB) resulted in good antimicrobial activity. We also determined the effect of SP extract and fractions on cytotoxicity, and chemopreventive effect on human cancer cells. The experiment was conducted to determine cytotoxicity of SP partition layers on HepG2, HeLa and MCF-7 cells by MTT assay. Among the various partition layers of SP, SPMEE and SPW were showed the strongest cytotoxic effects on all cancer cell lines. The quinone reductase induced activities of HepG2 cell, the butanol partition layer (SPMB) at a does of 40 $\mu\textrm{g}$/mL was 8.49 times more effective compared to the control value of 1.0. This value was significantly higher than that of previous results using the other materials. Therefore, based on these studies, SP may be developed into a potentially useful antimicrobial and anticarcinogenic agents.

An Isocoumarin with Hepatoprotective Activity in Hep G2 and Primary Hepatocytes from Agrimania pilosa

  • Park, Eun-Jeon;Oh, Hyun-Cheol;Kang, Tai-Hyun;Sohn, Dong-Hwan;Kim, Youn-Chul
    • Archives of Pharmacal Research
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    • v.27 no.9
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    • pp.944-946
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    • 2004
  • Phytochemical investigation of the aqueous extract of ~he roots of Agrimania pilosa Ledeb. (Rosaceae), as guided by hepatoprotective activity in vitro, furnished two isocoumarins, agri-monolide (1) and agrimonolide 6-O-$\beta$-D-glucoside (3), and (+)-catechin (2). Compound 1 showed hepatoprotective effects on both tacrine-induced cytotoxicity in human liver-derived Hep G2 cells and tert-butyl hydroperoxide-induced cytotoxicity in rat primary hepatocytes with EC$_{50}$ values of 88.2$\pm$2.8 and 37.7$\pm$1.6 $\mu$M, respectively.y.