• 제목/요약/키워드: HepG2 cell line

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유황오리 추출물로부터 항종양활성 성분의 분리 및 정제

  • 윤원호;황진용;김창한
    • 한국축산식품학회:학술대회논문집
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    • 한국축산식품학회 2004년도 정기총회 및 제33차 춘계 학술대회
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    • pp.373-375
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    • 2004
  • 본 연구는 유황오리로부터 항종양 효과를 나타내는 물질을 용매추출과 각종 chromatography를 사용하여 분리 및 정제하였다. HPLC를 사용하여 정제한 항종양활성 성분의 수율은 유황오리 1Kg당 10mg이었다. HPLC를 이용하여 정제한 항종양활성 물질을 Clonogenic assay로 측정한 결과 $200{\mu}g/m{\ell}$의 농도에서 HEp-2, KB 및 SW-156에서 각각 26%, 28%및 30%의 생존율을 나타내어 항종양 효과가 나타났다. MTT assay에서는 $100{\mu}g/m{\ell}$의 농도는 Farrow, HEp-2 및 KB에 대해서 각각 56%, 58% 및 56%의 세포증식 억제 효과를 나타내었다. bovine normal kidney cell line인 MDBK는 $500{\mu}g/m{\ell}$, $200{\mu}g/m{\ell}$, $100{\mu}g/m{\ell}$에서 각각 48%, 34%, 28%로 증식억제 효과가 나타나 정상세포에 대한 세포독성은 강하지 않은 것으로 판단되어진다.

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miR-340 Reverses Cisplatin Resistance of Hepatocellular Carcinoma Cell Lines by Targeting Nrf2-dependent Antioxidant Pathway

  • Shi, Liang;Chen, Zhan-Guo;Wu, Li-li;Zheng, Jian-Jian;Yang, Jian-Rong;Chen, Xiao-Fei;Chen, Zeng-Qiang;Liu, Cun-Li;Chi, Sheng-Ying;Zheng, Jia-Ying;Huang, Hai-Xia;Lin, Xiang-Yang;Zheng, Fang
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권23호
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    • pp.10439-10444
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    • 2015
  • Many chemotherapeutic agents have been successfully used to treat hepatocellular carcinoma (HCC); however, the development of chemoresistance in liver cancer cells usually results in a relapse and worsening of prognosis. It has been demonstrated that DNA methylation and histone modification play crucial roles in chemotherapy resistance. Currently, extensive research has shown that there is another potential mechanism of gene expression control, which is mediated through the function of short noncoding RNAs, especially for microRNAs (miRNAs), but little is known about their roles in cancer cell drug resistance. In present study, by taking advantage of miRNA effects on the resistance of human hepatocellular carcinoma cells line to cisplatin, it has been demonstrated that miR-340 were significantly downregulated whereas Nrf2 was upregulated in HepG2/CDDP (cisplatin) cells, compared with parental HepG2 cells. Bioinformatics analysis and luciferase assays of Nrf2-3'-untranslated region-based reporter constructor indicated that Nrf2 was the direct target gene of miR-340, miR-340 mimics suppressing Nrf2-dependent antioxidant pathway and enhancing the sensitivity of HepG2/CDDP cells to cisplatin. Interestingly, transfection with miR-340 mimics combined with miR-340 inhibitors reactivated the Nrf2 related pathway and restored the resistance of HepG2/CDDP cells to CDDP. Collectively, the results first suggested that lower expression of miR-340 is involved in the development of CDDP resistance in hepatocellular carcinoma cell line, at least partly due to regulating Nrf2-dependent antioxidant pathway.

Cytotoxic Activity of the Extracts from Curcuma zedoaria

  • Kim, Myoun-Gae;Kim, Jung-Sun;Hong, Jon-Ki;Ji, Ming-Jie;Lee, Yong-Kyu
    • Toxicological Research
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    • 제19권4호
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    • pp.293-296
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    • 2003
  • The effect of the hexane extract of Curcuma zedoaria roots and its solvent fractions were investigated on the proliferation of SiHa, SNU-1 and HepG2 cell lines. Among those fractions, final fraction H2-3-1 and H2-3-3 showed cytotoxic effect on SiHa and HepG2 cell lines. The hallmark of apoptosis, DNA fragmentation, also appeared in the final fractions H2-3-1 and H2-3-3 after 24h treatment in SiHa cell line. Furthermore, those fractions were shown to be able to induce cell death in $[^3H]$thymidine incorporation test. These two fractions, H2-3-1 and H2-3-3 were determined as (-)-$\alpha$-curcumene and $\beta$-tumerone respectively by NMR and mass spectrum. From these results, it is speculated that te hexane extract of Curcuma zedoaria is necessary for further studies as a potent inhibitor of the growth of cancer cells.

Interrelationship between Cell Differentiation and Expression of mRNA for Transferrin in HL-60 Leukemia Cell Line

  • Lee, Soo-Young;Chi, Chung-Hee;Kim, You-Mie
    • BMB Reports
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    • 제33권4호
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    • pp.308-311
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    • 2000
  • The interrelationship between the differentiation and expression of mRNA for transferrin in the HL-60 leukemia cell line was studied. Transferrin mRNA was expressed in HL-60 leukemia cells and the amount was 50% of that in the positive control cell line, HepG-2 cells. The expression of $T_f$ mRNA in HL-60 cells was not regulated by IL-1, IL-6 and $TNF-{\alpha}$, respectively. The expression of $T_f$ mRNA in the differentiated cells into a granulocyte lineage by DMSO, or all-trans RA, was up-regulated (160-170% of control cells); whereas, the expression was not regulated in the differentiated cells into a macrophage lineage by PMA. These results suggest that the differentiation to a granulocyte lineage of HL-60 leukemia cells appear to be related with the upregulation of transferrin mRNA expression.

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간암 치료 신약 개발 및 이의 제제화 연구 (II. 제제화 연구)

  • 정홍석;최명준;장진수;황유경;안교한;이석종
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
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    • pp.251-251
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    • 1994
  • 1차년도에서 이미 확립한 다양한리포좀의 제조 방법에 따라간세포를표 적하기 위해 이들 리포좀의 조성에 galactocercbroside를 함유시켜 targeted liposomes을 제조하였다. 표적리포좀의 제조 방법에 따른 봉합율 (cncapsulation)과 안정성을 조사했을 때, freezing-thawing 과정을 거친 리포좀이 encapsulation 과 stability가 가장 뛰어남을 알 수 있었다. In vitro 간세포 표적능력을 in vitro cell culture system에서 간세포 cell line (Hep G2, 2.2.15과 다른 cell line (Vero E6, J82)에 대한 표적리포좀과 control liposomes의 uptake를 FTTC 또는 CF 형광으로 측정했을 때, 표적리포좀이 간세포 cell line에 더 많이 uptake하는 것을 알았다. In vivo 실험에서는 6 - 8 주령의 mouse tail vain에 표적리포좀과 control liposomes을 주사하여 각 장기에 존재하는 리포좀의 양을 형광으로 측정했을 때, 표적리포좀 (targetcd liposomes)이 다른 장기에 비해 liver에 선택적으로 많이 분포함을 알수있었다. 또한 간암의 효과적인 치료를 위해 간세포와 암세포를 동시에 표적할 수는 double targeting liposomes을 design 하였다.

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Cytotoxic and antioxidant properties of four plants belonging to the genus Solanum

  • Dongre, Santoshkumar H.;Badami, Shrishailappa;Godavarthi, Ashok;S., Mahendran;P., Vijayan;B., Suresh
    • Advances in Traditional Medicine
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    • 제8권1호
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    • pp.86-92
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    • 2008
  • The aim of the study was to evaluate in vitro antioxidant and cytotoxic activities of the methanolic extracts of leaves of Solanum sisymbrifolium, Solanum anguivi multiflora, Solanum barbisetum and Solanum jasminoides. In the in vitro antioxidant screening using ABTS [2,2'-azino-bis (3-ethylbenzo-thiazoline-6-sulphonic acid) diammonium salt] method, the methanol extracts of Solanum jasminoides, Solanum barbisetum and Solanum anguivi multiflora exhibited potent antioxidant activity with $IC_{50}$ values 31.25 $\pm$ 0.35, 40.33 $\pm$ 0.57 and 54.33 $\pm$ 0.57 ${\mu}g$/ml, respectively. Solanum barbisetum also showed potent activity in DPPH [1,1-diphenyl-2-picryl hydrazyl] method with an $IC_{50}$ value of 55.33 $\pm$ 1.66 ${\mu}g/ml. In the cytotoxicity studies, the methanol extract of Solanum barbisetum exhibited moderate activity against Vero, HEp-2, HeLa and A-549 cell lines with $IC_{50}$ values in the range of 83.30 - 127.30 ${\mu}g$/ml. Solanum anguivi multiflora extract also showed moderate activity against Hep-2 cell line with $IC_{50}$ value of 80.13 ${\mu}g$/ml. Solanum barbisetum possessing both the activities requires further investigation.

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

  • 심정섭;김일환;김강산;강병기;최수덕
    • 대한한방내과학회지
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    • 제22권1호
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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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Conjugated Linoleic Acid Changes fatty Acid Composition by Decreasing Monounsaturated fatty Acids in Rabbits and Hep G2 Cells

  • Nam, Kisun
    • Journal of Nutrition and Health
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    • 제30권4호
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    • pp.442-450
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    • 1997
  • Conjugated dienoic derivatives of linoleic acid(CLA) are a mixture of positional and geometric isomers of linoleic acid(LA). We previously found that CLA changes the fatty acid profile in chicken eggs and serum by decreasing monounsaturated fatty acids. Studies were conducted to explore the effects of CLA on fatty acid composition. Rabbits were fed a semisynthetic diet with or without CLA(0.5g CLA/rabbit/day) for 22 weeks. Compared to the control, rabbits fed CLA had significantly lower monounsaturated fatty acid levels(palmitoleic acid Cl6 : 1 by 50% and oleic acid Cl8 : 1, by 20%) in plasma lipids. We found similar differences in fatty acid composition in the liver and the aorta. The inhibitory effect of CLA on $\Delta$9 desaturation was confirmed in a human hepatoma cell line, Hep G2. CLA significantly decreased $\Delta$9 desaturation in 4-5 hours as shown by an increase in the ratio of Cl6 : 0 to C 16 1, This is apparently due to a decrease in $\Delta$9 desaturase(stearoyl-CoA desaturase, SCD) activity ; it was decreased more than 50%. These results, along with our previous findings, indicate that CLA is an inhibitor of $\Delta$9 desaturase in the liver.

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변이 도파민 2 수용체와 나트륨 옥소 공동 수송체 이입유전자의 이중 리포터시스템 개발 (Development of Dual Reporter System of Mutant Dopamine 2 Receptor ($D_2R$) and Sodium Iodide Symporter (NIS) Transgenes)

  • 황도원;이동수;강주현;장영수;김윤희;정재민;정준기;이명철
    • 대한핵의학회지
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    • 제38권4호
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    • pp.294-299
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    • 2004
  • 목적 : 현재 생체 내로 이식된 세포를 추적하는데 여러 가지 리포터 유전자들이 이용되고 있다. 이 연구에서는 사람 나트륨 옥소 공동 수송체 (hNIS)와 도파민 2 수용체($D_2R$)를 이중 리포터 유전자로 사용하여 각각을 비교하였다. 대상 및 방법: hNIS와 $D_2R$의 발현이 동시에 이루어지도록 하기 위해서 IRES (Internal ribosome entry site)로 연결된 재조합 플라스미드(pIRES-hNIS/D_2R)를 제조하였다. $pIRES-hNIS/D_2R$를 사람의 간암세포주인 SK-Hep1에 lipofactamine을 이용하여 형질을 도입시킨 후, 항생제(G418)를 농도별로 처리하여 2주간 선별하였다(HEP-ND). hNIS와 $D_2R$발현 유무와 발현 정도를 알아보기 위하여 각 유전자에 특이적인 프라이머를 이용하여 RT-PCR을 수행하였다. 각 형질 도입세포군에서, hNIS의 활성은 $^{125}I$ 섭취율을 이용하여 측정하고 $D_2R$의 활성은 $[^3H]spiperone$을 리간드로 이용하여 수용체 결합 정도를 측정하였다. 결과: 선별된 HEP-ND세포에서 hNIS와 $D_2R$의 발현을 RT-PCR로 확인하였을 때 IRES로 연결된 hNIS와 $D_2R$의 발현 정도는 서로 비슷하였다. HEP-ND세포의 $^{125}I$ 섭취율은 대조군인 SK-Hep1세포에 배해 30-40배 증가되었고, $KClO_4$에 의해 $^{125}I$ 섭취가 저해되었다. $D_2R$의 발현 정도를 측정할 수 있는 수용체 결합 분석법을 통해G418 농도별로 나눈 두 종류의 세포주에서, $[^3H]spiperone$을 이용한 해리상수 ($K_d$)와 최대결합 부위농도 ($B_{max}$)는 각각 2.92 nM, 745.25 fmol/mg protein과 8.91nM, 1323 fmole/mg protein이었다. hNIS와 $D_2R$발현의 상관관계에서는 높은 상관관계를 나타내었다. 결론: 이 연구에서 hNIS와 $D_2R$가 이입된 세포주에서 이중 유전자, 감마 영상 리포터 시스템을 개발하였으며, $D_2R$와 HNIS 유전자를 이중 핵 영상 시스템으로서 서로 상호보완적으로 이용할 수 있을 것으로 기대하고 있다.

결명자의 타크린으로 유발한 간 세포독성 보호 성분 (Hepatoprotective compounds of Cassiae Semen on tacrine-induced cytotoxicity in Hep G2 cells)

  • 변에리사;정길생;안인파;리빈;이동성;고은경;윤권하;김윤철
    • 생약학회지
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    • 제38권4호
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    • pp.400-402
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    • 2007
  • Tacrine is an acetylcholinesterase inhibitor that is approved for the treatment of Alzheimer's disease. However, tacrine treatment for Alzheimer's disease results in reversible hepatotoxicity in 30-50% of patients, which seriously limits its clinical use. Accordingly, the identification of constituents in natural products that have protective effects on tacrine-induced hepatotoxicity would be valuable. In the present study, an immortalized human hepatoma cell line, HepG2 was employed to screen for agents that protect against tacrine-induced hepatotoxicity. The bioassay-guided fractionation of water extract of Cassiae Semen furnished two anthraquinones, aurantio-obtusin (1) and obtusifolin (2). Compounds 1 and 2 showed hepatoprotective effects with the protection ratio values of 55.3 +/- 0.5% and 41.2 +/- 0.8% at $160{\mu}M$, respectively.