• Title/Summary/Keyword: HepG2 2.2.15

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Inhibitory Effects of Eugenia caryophyllate, Ephedra sinica and Cinnamomum cassia on the Replication of HBV in HepG2 2.2.15 Cells (정향, 마황, 계피의 간염 B형 바이러스 증식 억제 효과)

  • 강석연;김태균;박민수;한형미;정기경;강주혜;문애리;김승희
    • Biomolecules & Therapeutics
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    • v.7 no.2
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    • pp.133-137
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    • 1999
  • This study was undertaken to test for anti-Hepatitis B virus (HBV) activity of the aqueous extracts prepared from Eugenia caryophyllate, Ephedra sinica, Cinnamomum cassia. Aqueous extracts were assayed for the inhibition of HBV replication by measurement of HBV DNA and surface antigen (HBsAg) levels in the extracellular medium of HePG2 2.2.15 cells. All extracts decreased the levels of extracellular HBV virion DNA at concentrations ranging from 128 to 256 $\mu$g/ml and inhibited the production of HBsAg dose-dependently. Our findings suggest that these three hebal medicinal plants may have potential to develop as specific anti-HBV drugs in the future.

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Platycodin D Induces Apoptosis, and Inhibits Adhesion, Migration and Invasion in HepG2 Hepatocellular Carcinoma Cells

  • Li, Ting;Xu, Wen-Shan;Wu, Guo-Sheng;Chen, Xiu-Ping;Wang, Yi-Tao;Lu, Jin-Jian
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.4
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    • pp.1745-1749
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    • 2014
  • Background: Platycodin D (PD), a triterpenoid saponin isolated from the Chinese medicinal herb Platycodonis radix, possesses anti-cancer effects in several cancer cell lines. The aim of this study was to evaluate its anticancer activities in hepatocellular carcinoma cells. Materials and Methods: MTT and colony formation assays were performed to evaluate cell proliferation, along with flow cytometry and Western blotting for apoptosis. Cell adhesion was tested by observing cellular morphology under a microscope, while the transwell assay was employed to investigate the cell migration and invasion. Results: PD concentration-dependently inhibited cell proliferation in both HepG2 and Hep3B cells, and significantly suppressed colony formation and induced apoptosis in HepG2 cells. The protein levels of cleaved poly ADP-ribose polymerase (PARP) and Bax were up-regulated while that of survivin was down-regulated after treatment with PD. Moreover, PD not only obviously suppressed the adhesion of HepG2 cells to Matrigel, but also remarkably depressed their migration and invasion induced by 12-O-tetradecanoylphorbol 13-acetate (TPA). Conclusions: PD presents anti-cancer potential in hepatocellular carcinoma cells via inducing apoptosis, and inhibiting cell adhesion, migration and invasion, indicating promising features as a lead compound for anti-cancer agent development.

Secretion of Inflammatory Cytokines by Aloe vera Extract in HepG2 Cells (HepG2 세포에서 알로에 베라 추출물에 의한 염증성 사이토카인 분비)

  • Kim, Ilrang
    • The Korean Journal of Food And Nutrition
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    • v.27 no.3
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    • pp.400-405
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    • 2014
  • Recently, cases of Aloe vera induced-toxic hepatitis have been reported. However, the precise inflammatory effects of Aloe vera extract have not been clearly elucidated yet. In this study, the inflammatory effects and the mechanism of 70% ethanolic Aloe vera extract on liver were evaluated by in vitro assays using human hepatoma HepG2 cells. Cell viability was investigated using MTT assay at $0.001{\sim}100{\mu}g/mL$ of Aloe vera extract. To evaluate inflammatory effect of the Aloe vera extract, the secretion of pro-inflammatory cytokine Interleukin 8 (IL-8) and Macrophage colony-stimulating factor (M-CSF) were detected. Aloe vera extract did not induce cell death at concentrations of $0.001{\sim}100{\mu}g/mL$. However, Aloe vera extract significantly increased the IL-8 secretion by 15.7~25.8% and the M-CSF secretion by 36.6~61.5% at the same concentrations. These results indicate that Aloe vera extract shows an inflammation-related mild hepatotoxicity than a severe toxicity such as cell death and this hepatitis is mediated by the secretion of inflammatory cytokine IL-8 and M-CSF.

The Effects on Antimicrobial and Cytotoxicity of Solanum Iyratum Fractions (배풍등 분획물의 항균 및 암세포 증식 억제효과)

  • Shin Mi-Ok
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.948-954
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    • 2005
  • In this study, we investigated antimicrobial and cytotoxicity effects to each fraction extracted from Solanum lyratum (SL), which were extracted methanol (SLM) and then the extract was fractionated into five different types : hexane (SLMH), ethyl ether (SLMEE), ethylacetate (SLMEA), butanol (SLMB) and aqueous (SLMA). The antimicrobial activity was analyzed by the paper disc method. Among the various solvent fractions, SLMEA showed the strongest antimicrobial activies. The cytotoxicity of SL fractions on HeLa, MCF-7, HT-29 and HepG2 cells was evaluated by MTT assay. Among various partition layers, SLMEE showed the strongest cytotoxic effects to all cancer cell lines. We also observed that quinone reductase (QR) was induced by all fraction layers of SL to HepG2 cells. Since the QR-induced effects of SLMEE on HepG2 cells at $160{\mu}g/ml$ concentration showed 2.1 when compared with a control value of 1.0, inducer of QR for cancer protection may be contained in this fraction.

Radioiodine Therapy of Liver Cancer Cell Following Tissue Specific Sodium Iodide Symporter Gene Transfer and Assessment of Therapeutic Efficacy with Optical Imaging (조직 특이 발현 Sodium Iodide Symporter 유전자 이입에 의한 방사성옥소 간암세포 치료와 광학영상을 이용한 치료효과 평가)

  • Jang, Byoung-Kuk;Lee, You-La;Lee, Yong-Jin;Ahn, Sohn-Joo;Ryu, Min-Jung;Yoon, Sun-Mi;Lee, Sang-Woo;Yoo, Jeong-Soo;Cho, Je-Yeol;Lee, Jae-Tae;Ahn, Byeong-Cheol
    • Nuclear Medicine and Molecular Imaging
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    • v.42 no.5
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    • pp.383-393
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    • 2008
  • Purpose: Cancer specific killing can be achieved by therapeutic gene activated by cancer specific promotor. Expression of sodium iodide symporter (NIS) gene causes transportation and concentration of iodide into the cell, therefore radioiodine treatment after NIS gene transfer to cancer cell could be a form of radionuclide gene therapy. luciferase (Luc) gene transfected cancer cell can be monitored by in vivo optical imaging after D-luciferin injection. Aims of the study are to make vector with both therapeutic NIS gene driven by AFP promoter and reporter Luc gene driven by CMV promoter, to perform hepatocellular carcinoma specific radiodiodine gene therapy by the vector, and assessment of the therapy effect by optical imaging using luciferase expression. Materials and Methods: A Vector with AFP promoter driven NIS gene and CMV promoter driven Luc gene (AFP-NIS-CMV-Luc) was constructed. Liver cancer cell (HepG2, Huh-7) and non liver cancer cell (HCT-15) were transfected with the vector using liposome. Expression of the NIS gene at mRNA level was elucidated by RT-PCR. Radioiodide uptake, perchlorate blockade, and washout tests were performed and bioluminescence also measured by luminometer in these cells. In vitro clonogenic assay with 1-131 was performed. In vivo nuclear imaging was obtained with gamma camera after 1-131 intraperitoneal injection. Results: A Vector with AFP-NIS-CMV-Luc was constructed and successfully transfected into HepG2, Huh-7 and HCT-15 cells. HepG2 and Huh-7 cells with AFP-NIS-CMV-Luc gene showed higher iodide uptake than non transfected cells and the higher iodide uptake was totally blocked by addition of perchlorate. HCT-15 cell did not showed any change of iodide uptake by the gene transfection. Transfected cells had higher light output than control cells. In vitro clonogenic assay, transfected HepG2 and Huh-7 cells showed lower colony count than non transfected HepG2 and Huh-7 cells, but transfected HCT-15 cell did not showed any difference than non transfected HCT-15 cell. Number of Huh-7 cells with AFP-NIS-CMV-Luc gene transfection was positively correlated with radioidine accumulation and luciferase activity. In vivo nuclear imaging with 1-131 was successful in AFP-NIS-CMV-Luc gene transfected Huh-7 cell xenograft on nude mouse. Conclusion: A Vector with AFP promoter driven NIS and CMV promoter driven Luc gene was constructed. Transfection of the vector showed liver cancer cell specific enhancement of 1-131 cytotoxicity by AFP promoter, and the effect of the radioiodine therapy can be successfully assessed by non-invasive luminescence measurement.

Effects of Constitutive Androstane Receptor (CAR) on PBRU Transactivation of CYP2B Gene in Different Culture Cell Types: Comparison Between Hep G2 and COS-cells (배양세포의 Type에 따른 Constitutive Androstane 수용체 (CAR)의 CYP2B PBRU 전사활성 효과: Hep G2와 COS 세포의 비교)

  • 민계식
    • Journal of Life Science
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    • v.13 no.3
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    • pp.324-332
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    • 2003
  • The objective of this study was to examine if transient transfection of CAR can transactivate CYP2B1 PBRU reporter gene in COS cells in which the endogenous CYP2B1 gene is not induced by PB. In non-transfeced cells of both Hep G2 and COS, the endogeneous expression of CAR was not detected by antibody against CAR. When cultured cells were transfected with CAR expression plasmid, mCAR1-GFP, both cell types expressed high levels of CAR protein and could allow to examine the effect of CAR in PBRU transactivation. Both cell types expressed endogenous RXR and transfection of RXR expression plasmid dramatically increased its protein expression. Whereas CAR transactivated PBRU2C1Luciferase about 12 fold as compared to 2C1Luciferase in Hep G2 cells, it did not stimulate the luciferase activity of the PBRU reporter gene in COS cells. These results indicate that Hep G2 cells can respond to CAR differently from COS cells, and suggest that factors other than CAR and RXR may be required in inducing PBRU activation and the expression of these factors may be different between liver and kidney.

Inhibition of HBV ]Replication by the Extract of Phyllanthus ussuriensis (여우주머니(Phyllanthus ussuriensis) 추출물의 B헝 간염바이러스 증식 억제)

  • Kim, Chul-Young;Kim, Jeong-Min;Kim, Tae-Gyun;Kim, Seung-Hee;Huh, Hoon
    • Biomolecules & Therapeutics
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    • v.6 no.2
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    • pp.139-144
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    • 1998
  • Abstract -Phyllanthus ussuriensis which belongs to Euphorbiaceae has been used as a component of Korean traditional remedy against hepatitis and jaundice. Aqueous methanolic extract of P. ussuriensis was tested for antiviral activity of hepatitis B virus (HBV) in HepG2 2.2.15 cells which were derived through transfection of cloned HBV DNA into HepG2 human hrpatoblastoma cells. P. ussuriensis extract decreased the levels of extracellular HBV virion DNA and inhibited HBV replication at concentrations ranging from 50 to 500$\mu$g/ml. Partitioning of water suspension of the extract revealed that the activity mainly resides in the EtOAc fraction. Consecutive purification of the EtOAc fraction by silica-gel column chromatography resulted in the two active anti-HBV fractions. Southern blot analysis shows that the action mechanisms or active site of the two fractions seems to be different in terms of their inhibition of HBV replication steps.

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Parthenolide-Induced Apoptosis, Autophagy and Suppression of Proliferation in HepG2 Cells

  • Sun, Jing;Zhang, Chan;Bao, Yong-Li;Wu, Yin;Chen, Zhong-Liang;Yu, Chun-Lei;Huang, Yan-Xin;Sun, Ying;Zheng, Li-Hua;Wang, Xue;Li, Yu-Xin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.12
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    • pp.4897-4902
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    • 2014
  • Purpose: To investigate the anticancer effects and underlying mechanisms of parthenolide on HepG2 human hepatocellular carcinoma cells. Materials and Methods: Cell viability was assessed by MTT assay and cell apoptosis through DAPI, TUNEL staining and Western blotting. Monodansylcadaverin(MDC) and AO staining were used to detect cell autophagy. Cell proliferation was assessed by Ki67 immunofluorescence staining. Results: Parthenolide induced growth inhibition in HepG2 cells. DAPI and TUNEL staining showed that parthenolide could increase the number of apoptotic nuclei, while reducing the expression of the anti-apoptotic protein Bcl-2 and elevating the expression of related proteins, like p53, Bax, cleaved caspase9 and cleaved caspase3. Parthenolide could induce autophagy in HepG2 cells and inhibited the expression of proliferation-related gene, Ki-67. Conclusions: Parthenolide can exert anti-cancer effects by inducing cell apoptosis, activating autophagy and inhibiting cell proliferation.

Antioxidant Effect of Cynomorii Herba on HepG2 Cells and Diphenyl-picryl-hydrazyl (DPPH) Radical Scavenging Activity (쇄양(鎖陽)의 Diphenyl-picryl-hydrazyl (DPPH) 소거 활성 및 HepG2 세포에 대한 항산화 효과)

  • Chang, Mun-Seog;Yang, Woong-Mo;Kim, Do-Rim;Park, Eun-Hwa;Park, Soo-Yeon;Park, Seong-Kyu
    • Herbal Formula Science
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    • v.15 no.2
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    • pp.139-145
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    • 2007
  • The purpose of this study was to investigate the anti-oxidant effect of Cynomorium songaricum. The extract of Cynomorii Herba was studied for diphenyl-picryl-hydrazyl (DPPH) radical scavenging activity, HepG2 cell viability and $H_2O_2$-induced cytotoxicity by a modified MTT assay. DPPH radical scavenging activity was measured after 30 minutes. The extract was tested by 1. 5, 10, 50, 100 and 500 ${\mu}g/ml$ concentrations. HepG2 cell viability by a modified MTT assay was measured in the concentrations of 10, 50, 100, 250, 500 ug/ml for 24 h. The results showed that the extract scavenged DPPH radical up to 52.2% with 50 ug/ml concentration. The extract did not reduced the cell viability and $H_2O_2-induced$ cytotoxicity (69.4%) was blocked by the extract in the concentrations of 50, 100, 250 and 500 ${\mu}g/ml$. In conclusion, the extract of Cynomorii Herba has potent antioxidant activity.

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Study on the Anti-Cancer Effect of Acanthopanax Sessiliflorus (오가피(五加皮)의 항암효과에 관한 연구)

  • Kim, Young-Chul;Woo, Hong-Jung;Lee, Jang-Hoon
    • The Journal of Korean Medicine
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    • v.20 no.3 s.39
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    • pp.54-65
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    • 1999
  • Objectives: Hepatoma is a very serious disease in Korea and vvorldwiclc. Hepatitis B vims (HBV) has proved the most significant cause of hepatoma. We canied out this study to investigate the effect of Acanthopanax sessilifloms on inhibiting cell proliferation and DNA synthesis in HepG2.2.15 cell line and on inhibiting phosphorilation of oncogene (MAP kinase) in NIT/3T3-HBx ceIl. Methods: To investigate the anti-cancer effect of Acanthopanax sessiliflorus, we did the CellTiter 96 Aqueous Non-radioactive Cell Proliferation assay (Promega); MTS/PMS assay, [$^3H$]-thymicline incorporation assay, and we measured the gene expression through westem blotting. Results: Acanthopanax sessiliflorus showed an inhibiting effect on the increase of HepG2.2.15 in the NTS/PMS assay. It also showed an inhibiting effect on DNA synthesis of HepG2.2.15 in the [$^3H$]-thymidine incorporation assay. Acanthopanax sessiliflorus showed an inhibiting effect of phosphorilation of MAP kinase in HBV - X genes. too. Conclusions: The results suggested that this herb had an anti cancer effect. We may discover an effective anti-cancer herb medicine through further studies on this herb medicine.

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