• Title/Summary/Keyword: Hep-G2 cell

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Inhibition of Diacylglycerol Acyltransferase by Phenylpyropenes Produced by Penicillium griseofulvum F1959

  • Lee, Seung-Woong;Rho, Mun-Chual;Choi, Jung-Ho;Kim, Koan-Hoi;Choi, Yong-Seok;Lee, Hyun-Sun;Kim, Young-Kook
    • Journal of Microbiology and Biotechnology
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    • v.18 no.11
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    • pp.1785-1788
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    • 2008
  • Phenylpyropenes A, B, and C, isolated from Penicillium griseofulvum F1959, inhibited DGAT in rat liver microsomes with $IC_{50}$ values of $78.7{\pm}1.6$, $21.7{\pm}0.2$, and $11.04{\pm}0.2{\mu}M$, respectively. In addition, a kinetic analysis using a Lineweaver-Burk plot revealed that phenylpyropene C was a noncompetitive inhibitor of DGAT. The apparent Michaelis constant ($K_m$) value and inhibition constant ($K_i$) value were calculated to be $8{\mu}M$ and $10.48{\mu}M$, respectively. Moreover, phenylpyropene C inhibited triglyceride formation in HepG2 cells.

Novel Trimeric Complex for Efficient Uptake of Plasmid Vector into HepG2 Cells

  • Joo, Jong-Hyuck;Park, Jong-Gu
    • Biomedical Science Letters
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    • v.9 no.2
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    • pp.67-74
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    • 2003
  • Viral and non-viral vectors have been used in the delivery of genetic materials into animal cells and tissues, with each approach having pros and cons. Non-viral vectors have many useful merits such as easy preparation, low immunity and size tolerance of a transgene when compared to those of viral vectors. Delivery specificity may be achieved by complex formation between receptor ligands and a non-viral vector. In the present study, non-viral vector systems are investigated in an effort to find a practical delivery means for gene therapy, Receptor-ligand interaction between transferrin-receptor and transferrin was utilized for efficient gene transfer into cancer cells. A plasmid vector, pcDNA3 (LacZ) was ligated with a small duplexed oligo fragment in which a Biotin- VN$^{TM}$ phosphoramidite was placed in the middle of the oligo. The plasmid vector labeled by biotin was then conjugated with biotin-labeled transferrin via streptavidin. This trimeric conjugates were delivered to a hepatoma cell line, HepG2. The delivery efficiency of the trimeric conjugate was 2-fold higher than that of cationic liposomes used for transfection of a plasmid vector. These results demonstrate that a plasmid vector can be efficiently transferred into cells by forming a trimeric complex of plasmid vector-linker-ligand.

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Effects of Purple Kohlrabi (Brassica oleracea var. gongylodes) Flesh and Peel Ethanol Extracts on the Antioxidant Activity and Antiproliferation of Human Cancer Cells (자색 콜라비 가식부와 껍질 에탄올 추출물의 항산화 활성 및 암세포 증식 억제효과)

  • Yang, Myung-Ja;Cha, Seon-Suk;Lee, Jae-Joon
    • The Korean Journal of Community Living Science
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    • v.26 no.2
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    • pp.405-414
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    • 2015
  • This study examines the effects of purple Kohlrabi fresh and peel ethanol extracts on the antioxidative activity and antiproliferation of human cancer cells (Hep G2 human liver, HCT-116 human colon, and A549 human lung cancer cells.) The total flavonoid and anthocyanin content of purple Kohlrabi ethanol extracts were much greater in the peel than in the flesh. The DPPH radical scavenging activity and antioxidative index of purple Kohlrabi peel extracts were similar to those of the BHA and the BHT. Antiproliferation effects of purple Kohlrabi peel extracts on human cancer cells (Hep G2, HCT-116, and A549) strengthened in a dose-dependent manner. In particular, the antiproliferation activity of purple Kohlrabi peel extracts exceeded 40% in colon cancer cells. These results indicate that the purple Kohlrabi peel may contain bioactive compounds such as flavonoids as well as anthocyanin and that these compounds may facilitate cancer prevention.

Ibuprofen Increases the Hepatotoxicity of Ethanol through Potentiating Oxidative Stress

  • Kim, Minjeong;Lee, Eugenia Jin;Lim, Kyung-Min
    • Biomolecules & Therapeutics
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    • v.29 no.2
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    • pp.205-210
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    • 2021
  • Over 30 million prescriptions of NSAIDs (non-steroidal anti-inflammatory drugs) are issued every year. Considering that these drugs are available without a prescription as over the counter (OTC) drugs, their use will be astronomical. With the increasing use of NSAIDs, their adverse effects are drawing attention. Especially, stomach bleeding, kidney toxicity, liver toxicity, and neurological toxicity are reported as common. Ibuprofen, one of the extensively used NSAIDs along with aspirin, can also induce liver toxicity, but few studies are addressing this point. Here we examined the liver toxicity of ibuprofen and investigated whether co-exposure to ethanol can manifest synergistic effects. We employed 2D and 3D cultured human hepatoma cells, HepG2 to examine the synergistic hepatotoxicity of ibuprofen and alcohol concerning cell viability, morphology, and histology of 3D spheroids. As a result, ibuprofen and alcohol provoked synergistic hepatotoxicity against hepatocytes, and their toxicity increased prominently in 3D culture upon extended exposure. Oxidative stress appeared to be the mechanisms underlying the synergistic toxicity of ibuprofen and alcohol as evidenced by increased production of ROS and expression of the endogenous antioxidant system. Collectively, this study has demonstrated that ibuprofen and EtOH can induce synergistic hepatotoxicity, providing a line of evidence for caution against the use of ibuprofen in combination with alcohol.

Effect of Injin Butanol Fraction with Thin Layer Chromatography on Fas-mediated Apoptosis (인진butanol 분획의 TLC추출성분이 Fas-mediated Apoptosis에 미치는 영향)

  • 박용진;김영철;이장훈;우흥정
    • The Journal of Korean Medicine
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    • v.23 no.2
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    • pp.57-69
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    • 2002
  • Objective: The aim of this study is to investigate the effect of Injin butanol fractions with Thin Layer Chromatography on Fas-mediated Apoptosis. Method: Injin-butanol fraction separated by TLC. MIT assay, cell cycle analysis, Caspase-3 protease assay, DNA fragmentation assay and quantitative RT-PCR were performed to evaluate the effects of TLC extraction of lnjin-butanol fraction on cell viability, cell cycle progression and apoptosis. Results: Scopoletin, luteolin, apigenin and unknown powder was isolated by TLC. Fas-mediated apoptosis analysis shows that scopoletin has inhibiting function on apoptosis. Caspase- 3 protease assay analysis shows that scopoletin inhibits activity of caspase-3. Quantitative RT-PCR analysis shows that no activity on caspase-3, but apoptosis inhibition cytokine -Bcl-2- is activated, and apoptosis activating cytokine -Bax- is unactivated. Conclusion: These results show that each fraction of Injin-butanol TLC extraction, especially scopoletin, acts as a protective function on liver cell viability, and inhibitory function on apoptosis. (J Korean Oriental Moo 2002;23(2):57-69)

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Effect of Endothelial Cell Growth Factor and Cyclic AMP Increasers on the Proliferation of Human Omental Microvascular Endothelial Cells (사람의 대망미세혈관내피세포 증식에 대한 내피세포성장인자 및 CYCLIC AMP 증가물질의 효과)

  • Kim, Won-Gon;Kim, Jong-Man;Yu, Se-Yeong
    • Journal of Biomedical Engineering Research
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    • v.16 no.4
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    • pp.463-470
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    • 1995
  • Complete prelining of artificial vascular grafts with autologous endothelial cells may be one of the ideal solutions to obtain a nonthrombogenlc blood-contacting surface. To establish an intact endothelial cell monolayer on a prosthetic surface at the time of implantation,a sufficient number of endothelial cells and adequate propagation condition In cell culture are prerequisites. In this experimental study, endothelial cells from microvessels of adult human oriental adipose tissue were enzymatically harvested, and optimal culture conditions for proliferation of the endothelial cells in cell culture were examined. Human oriental adipose tissue was digested with collagenase and endothelial cells were separated from other stromal elements by mesh filtration method. Cultured cells were identified as endothelial cells by immunofluorescent staining for factor VIII-related antigen. Proliferation in usual 20% fetal bovine serum (FBS) medium or medium containing endothelial cell growth factor (ECGF)(5 ng/ml) and heparin (HEP)(1,000 units/ml) were compared,and the effects of adding compounds that increase intracellular cyclic adenosine monophosphate levels, that is,cholera toxin (CT)(1 $\mu\textrm{g}$/ml) and isobutylmethylxanthine (IBMX)(0.2 ml),were also analyzed. In total,following eight media groups were examined. 1) FBS medium + ECGF + HEP, 2) FBS medium + ECGF + HEP+CT, 3) FBS medium+ECGF+HEP+lBMX, 4) FBS medium+ECGF+HEP+CT+ IBMX, 5) FBSmedium, 6) FBS medium +CT, 7) FBS medium + IBMX, 8) FBS medium + CT + IBMX. It was shown that the medium containing ECGF + HEP with or without cholera toxin was most efficient in Stimulating cell proliferation. IBMX was considered to have antagonistic effect to ECGF. Among experimental groups without ECGF and HEP, the addition of cholera toxin and IBMX was shown to significantly potentiate cell proliferation. This results could provide a practical method for use of cultured human endothelial cells for endothelial cell seeding of cardiovascular prosthetic device, particularly in small-diameter vascular grafts.

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Microbiological Characteristics and Cytoprotective Effects of Samjung-Hwan Fermented by Lactic Acid Bacteria (유산균을 이용한 발효삼정환의 미생물 특성 및 세포 보호 효과)

  • Chang, Seju;Wang, Jing-Hua;Shin, Na Rae;Kim, Hojun
    • Journal of Korean Medicine for Obesity Research
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    • v.16 no.1
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    • pp.11-18
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    • 2016
  • Objectives: To confirm microbiological change and cytoprotective effect of Samjung-hwan (SJH) which fermented by Lactic acid bacteria from natural fermented SJH. Methods: SJH was fermented by Lactobacillus brevis and Lactococcus lactis subsp. lactis from natural fermented SJH. After 1 week of fermentation, we analysed pH and microbial profiling. We also performed measuring total polyphenol total flavonoid contents and 1,1-Diphenyl-2-picryhydrazyl (DPPH) free radical scavenging activity to investigate antioxidant ability. Cell viability was performed by using HepG2 cell. Results: pH of lactic acid bacteria inoculated group and non-inoculated group was decreased and total counts of lactic acid bateria for both group was increased after fermentation of SJH. Total polyphenol and flavonoid contents and DPPH free radical scavenging activity was increased in both group. Total polyphenol contents of lactic acid bacteria Inoculated group is more increased than non-inoculated group. HepG2 cell viability was increased in both group. Conclusions: SJH fermentd by Lactobacillus brevis and Lactococcus lactis subsp. lactis shows change in microbiological character and has cytoprotective effect. Further studies are required for investigating function of lactic acid bacteria during fermentation of SJH.

Hepatocyte protection and antioxidant effect of Citri Unshius Pericarpium against cadmium-induced oxidative stress (카드뮴으로 유발된 산화 스트레스에 대한 진피의 간세포 보호 및 항산화 효과)

  • Noh, Gyu Pyo;Byun, Sung Hui;Jung, Dae Hwa;Lee, Jong Rok;Park, Sook Jahr;Kim, Sang Chan
    • Herbal Formula Science
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    • v.28 no.4
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    • pp.327-337
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    • 2020
  • Objective : Citri Unshius Pericarpium is the dried peel of mature fruit of Citrus unshiu Markovich and has been used to treat indigestion, vomiting, and removal of phlegm. This study investigated the hepatoprotective and antioxidant effect of CEE (Ethanol extract of Citri Unshius Pericarpium) in cadmium (CdCl2)-treated HepG2 cells. Methods : Component analysis of Citri Unshius Pericarpium was analyzed by UPLC with C18 column. Cell viability was determined by MTT assay. The enzyme activity of superoxide dismutase (SOD) and the level of reactive oxygen species (ROS) and reduced glutathione (GSH) were analyzed using commercially available kits. Results : Cadmium caused severe HepG2 cell death. Cadmium also increased ROS production, consistent with depletion of GSH and inhibition of the SOD enzyme. However, CEE treatment reduced cell death and relieved oxidative stress caused by cadmium toxicity. CEE lowered ROS levels and improved depletion of GSH levels. CEE also enhanced the enzymatic activity of SOD. In component analysis, hesperidin was the most abundant of the five marker compounds (Narigenin, Narigin, Narirutin, Hesperidin and Hesperidin), which assumes that hesperidin partially contributed to the antioxidant activity of CEE. Conclusion : These results suggested that CEE could be a potential substance to solve heavy metal-related health problems. In particular, inhibition of oxidative stress by CEE can be a way to treat liver damage caused by cadmium.

Strobilanthes crispus Juice Concentrations and Anticancer Effects on DNA Damage, Apoptosis and Gene Expression in Hepatocellular Carcinoma Cells

  • Hussin, Faridah;Eshkoor, Sima Ataollahi;Rahmat, Asmah;Othman, Fauziah;Akim, Abdah;Eshak, Zolkapli
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.14
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    • pp.6047-6053
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    • 2015
  • Background: Hepatocellular carcinoma is one of the most common cancers worldwide. Its prevalence is increasing in many countries. Plant products can be used to protect against cancer due to natural anticancer and chemopreventive constituents. Strobilanthes crispus is one of plants with potential chemopreventive ability. Objective: This study aimed to evaluate the anticancer effects of Strobilanthes crispus juice on hepatocellular carcinoma cells. Materials and Methods: MTT assays, flow cytometry, comet assays and the reverse transcription-polymerase chain reaction (RT-PCR) were used to determine the effects of juice on DNA damage and cancer cell numbers. Results: This juice induced apoptosis after exposure of the HepG2 cell line for 72 h. High percentages of apoptotic cell death and DNA damage were seen at the juice concentrations above 0.1%. It was found that the juice was not toxic for normal cells. In addition, juice exposure increased the expression level of c-myc gene and reduced the expression level of c-fos and c-erbB2 genes in HepG2 cells. The cytotoxic effects of juice on abnormal cells were in dose dependent. Conclusions: It was concluded that the Strobilanthes crispus juice may have chemopreventive effects on hepatocellular carcinoma cells.

miR-101 Inhibiting Cell Proliferation, Migration and Invasion in Hepatocellular Carcinoma through Downregulating Girdin

  • Cao, Ke;Li, Jingjing;Zhao, Yong;Wang, Qi;Zeng, Qinghai;He, Siqi;Yu, Li;Zhou, Jianda;Cao, Peiguo
    • Molecules and Cells
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    • v.39 no.2
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    • pp.96-102
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    • 2016
  • miR-101 is considered to play an important role in hepatocellular carcinoma (HCC), but the underlying molecular mechanism remains to be elucidated. Here, we aimed to confirm whether Girdin is a target gene of miR-101 and determine the tumor suppressor of miR-101 through Girdin pathway. In our previous studies, we firstly found Girdin protein was overexpressed in HCC tissues, and it closely correlated to tumor size, T stage, TNM stage and Edmondson-Steiner stage of HCC patients. After specific small interfering RNA of Girdin was transfected into HepG2 and Huh7.5.1 cells, the proliferation and invasion ability of tumor cells were significantly inhibited. In this study, we further explored the detailed molecular mechanism of Girdin in HCC. Interestingly, we found that miR-101 significantly low-expressed in HCC tissues compared with that in matched normal tissues while Girdin had a relative higher expression, and miR-101 was inversely correlated with Girdin expression. In addition, after miR-101 transfection, the proliferation, migration and invasion abilities of HepG2 cells were weakened. Furthermore, we confirmed that Girdin is a direct target gene of miR-101. Finally we confirmed Talen-mediated Girdin knockout markedly suppressed cell proliferation, migration and invasion in HCC while downregulation of miR-101 significantly restored the inhibitory effect. Our findings suggested that miR-101/Girdin axis could be a potential application of HCC treatment.