• Title/Summary/Keyword: HepG2 cell line

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HepG2 세포의 산화적 손상에 대한 산삼 추출물의 보호효과 - DNA chip을 이용하여 -

  • Kim, Hyung-Seok;Park, Hee-Soo;Kwon, Ki-Rok
    • Journal of Pharmacopuncture
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    • v.10 no.1 s.22
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    • pp.121-135
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    • 2007
  • Objectives : This study was carried out to examine protective effect of wild ginseng extract on HepG2 human hepatoma cell line against tert-Butyl hydroperoxide (t-BHP)-induced oxidative damage. Methods : To evaluate protective effect of wild ginseng extract against t-BHP induced cytotoxicity, LDH level and activity of glutathione peroxidase and reductase were measured. Gene expression was also measured using DNA microarray. Results : Wild ginseng extract showed a significant protective effect against t-BHP-induced cytotoxicity in HepG2 cell line. It is not, however, related with the activities of glutathione peroxidase and glutathione reductase. Analysis of gene expression using DNA chip, demonstrated that 28 genes were up-regulated in t-BHP only group. Five genes - selenoprotein P, glutathione peroxidase 3, sirtuin 2, peroxiredoxin 2, serfiredoxin 1 homolog - may be related with the protective effect of wild ginseng extract. Conclusions : Based on the results, a protective effect of wild ginseng extract against t-BHP-induced oxidative damage in HepG2 cell line is not associated with the activities of glutathione peroxidase and glutathione reductase, but with the expression of selenoprotein P, glutathione peroxidase 3, sirtuin 2, peroxiredoxin 2, and serfiredoxin 1 homolog.

Cytotoxicity of Ligularia fischeri Extracts (곰취 추출물의 세포독성 효과)

  • 함승시;이상영;오덕환;정성원;김상헌;정차권;강일준
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.5
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    • pp.987-992
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    • 1998
  • This study was investigated to observe the cytotoxicity effect of Ligularia fischeri extracts against cancer cell lines including human lung carcinoma(A549), human cervix epitheloid carcinoma(HeLa) and human hepatocellular carcinoma(HepG2) using SRB(sulforhodamine B) method. The ethanol and methanol extracts of 1$\mu\textrm{g}$/${mu}ell$ showed approximately 79.2% and 86.4% cytotoxicity effects on HepG2 cell line and the ethyl acetate fracton fractionated from ethanol extracts showed the strongest cytotoxicity effect with 94% inhibition. The inhibitory effect of ethanol extract on HeLa cell line was somewhat low with 50~56% inhibition, but ethyl acetate fraction showed higher cytotoxicity effect with 91% and 91.9% inhibition on the HeLa and A549 cell line. On the contrary, the ethanol and methanol extracts showed the lower inhibition effects on the normal liver cell, WRL68, compared to human cancer cell lines.

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Selective in vitro cytotoxicity of Hypericum hookerianum towards cancer cell lines

  • P., Vijayan;S., Vinod Kumar;Badami, Shrishailappa;PK, Mukherjee;SA, Dhanaraj;B., Suresh
    • Advances in Traditional Medicine
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    • v.3 no.3
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    • pp.141-146
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    • 2003
  • The methanol extracts of the aerial parts, leaves and stem of Hypericum hookerianum were tested for in vitro cytotoxicity on selected normal and cancer cell lines and anti tumor activity using DLA cells. Cell viability and morphological changes were assessed. Among the three extracts tested, the stem extract of Hypericum hookerianum showed potent cytotoxicity against HEp-2 and RD cell lines. The $CTC_{50}$(concentration required to reduce viability by 50%) of this extract was found to be $2.02\;{\mu}g/ml$ for RD cell line, $10.25\;{\mu}g/ml$ for HEp-2 cell line and $100.06\;{\mu}g/ml$ for Vero cell line. In the clonogenic assay, no colony formation was observed up to a concentration of $100\;{\mu}g/ml$. In the short term cytotoxicity studies using DLA cells, 50% viability was observed in the concentration range of $50-100\;{\mu}g/ml$ for aerial parts, $100-200\;{\mu}g/ml$ for stem and more than $200\;{\mu}g/ml$ for leaf extracts of Hypericum hookerianum. In the long-term activity using HEp-2 cell line, no colony formation was observed over a concentration of 200 mg/ml for the stem extract. Hypericum hookerianum stem extract was fractionated into petroleum ether, chloroform, ethyl acetate and methanol soluble fractions. The petroleum ether and chloroform soluble fractions showed higher cytotoxic activity against HEp-2 cell line when compared to the other two fractions. The methanol stem extract of Hypericum hookerianum has the potential for further investigation in animal models to determine its anti-tumor activity and to identify its active principles.

mRNA Expression of Bax, Bcl-2, p53, Cathepsin B, Caspase-3 and Caspase-9 in the HepG2 Cell Line Following Induction by a Novel Monoclonal Ab Hep88 mAb: Cross-Talk for Paraptosis and Apoptosis

  • Mitupatum, Thantip;Aree, Kalaya;Kittisenachai, Suthathip;Roytrakul, Sittiruk;Puthong, Songchan;Kangsadalampai, Sasichai;Rojpibulstit, Panadda
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.2
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    • pp.703-712
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    • 2016
  • Monoclonal antibodies with specific antigens have been widely used as targeted therapy for cancer. Hep88 mAb is a monoclonal antibody which shows specific binding with anti-cancer effects against the HepG2 cell line. However, its mechanisms of action are still not completely understood. We examined cell cycling and apoptosis by flow cytometry and mRNA expression of factors involved in apoptosis and paraptosis in Hep88 mAb-treated HepG2 cells by real-time PCR. The cell-cycle analysis demonstrated that growth-inhibitory activity was associated with G2/M cell cycle arrest. Hep88 mAb induced a significant increase in apoptotic cell populations in a dose- and time-dependent manner. The mRNA expression results also suggested that the process triggered by Hep88 mAb involved up-regulation of tumor suppressor p53, pro-apoptotic Bax, Cathepsin B, Caspase-3 and Caspase-9, with a decrease of anti-apoptotic Bcl-2 - thus confirming paraptosis and apoptosis programmed cell death. These findings represent new insights into the molecular mechanisms underlying the anti-cancer properties of Hep88 mAb in liver cancer cells.

G1 Arrest of the Cell Cycle by Onchungeum in Human Hepatocarcinoma Cells (온청음(溫淸飮)이 인체 간암세포의 세포주기 G1 Arrest에 미치는 영향)

  • Goo, In-Moo;Shin, Heung-Mook
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.4
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    • pp.821-828
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    • 2008
  • Onchungeum, a herbal formula, which has been used for treatment of anemia due to bleeding, discharging blood and skin disease. In the present study, it was examined the effects of extract of Onchungeum (OCE) on the growth of human hepatocarcinoma cell lines Hep3B (p53 null type) and HepG2 (p53 wild type) in order to investigate the anti-proliferative mechanism by OCE. Treatment of Hep3B and HepG2 cells to OCE resulted in the growth inhibition in a dose-dependent manner, however Hep3B cell line exhibited a relatively strong anti-proliferative activity to OEC. Flow cytometric analysis revealed that OCE treatment in Hep3B cells caused G1 phase arrest of the cell cycle, which was associated with various morphological changes in a dose-dependent fashion. RT-PCR and immunoblotting data revealed that treatment of OCE caused the down-regulation of cyclin D1 expression, however the levels of cyclin E expression were not changed by OCE. The G1 arrest of the cell cycle was also associated with the induction of Cdk inhibitor p27 by OCE. Because the p53 gene is null in Hep3B cells, it is most likely that the induction of p21 is mediated through a p53-independent pathway. Moreover, p27 detected in anti-Cdk4 and anti-Cdk2 immunoprecipitates from the OCE-treated cells, suggesting that OCE-induced p27 protein blocks Cdk kinase activities by directing binding to the cyclin/Cdk complexes. Furthermore, OCE treatment potently suppresses the phosphorylation of retinoblastoma proteins and the levels of the transcription factor E2F-1 expression. Taken together, these results indicated that the growth inhibitory effect of OCE in Hep3B hepatoma cells was associated with the induction of G1 arrest of the cell cycle through regulation of several major growth regulatory gene products.

The Effect of Herbs on Inhibition of HBeAg Production in HepG2.2.15 Cell line (수종의 한약재가 HepG 2.2.15 Cell의 HBeAg발현 억제에 미치는 효과(效果))

  • Woo, Hong-Jung;Lee, Jang-Hoon;Kim, Young-Chul
    • The Journal of Internal Korean Medicine
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    • v.20 no.1
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    • pp.122-132
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    • 1999
  • Purpose : Hepatitis B virus DNA transfected cell line(HepG2.2.15) was cultured to evaluate the effect of herbs on the expression of HBeAg and the replication of HBV. HepG2.2.15 produces HBV particles as well as viral proteins into cell culture media. Methods : Extracts of herbs were adminitered to the cells on the proper concentration. Culture media was collected 48 hours after the herbal administration and HBeAg level in the media was examined by ELISA method. To confirm that the anti-viral effect was not due to direct cytotocixity of the extracts, normal cell proliferation was shown by cell counting. And as of the interference in protein synthesis of HepG2.2.15 by herb-extracts, we used the result of study that we performed before by ${\alpha}FP$ assay using EIA method. Results& Conclusion : Herb medicines like 地楡(Sanguisorbae Radix) and 覆盆子(Rubi Frusctus) showed significant inhibitory effect on HBeAg expression at p<0.01 and 五味子(Acanthopanacis Cortex) at p<0.05. Whereas, though some herbs such as ?草根(Rubiae Radix), 山査(Crataegii Fructus), 白芍藥(Paeoniae Radix Alba), and 大黃(Rhei Radix et Rhizoma) showed the tendecy to suppress HBeAg. most of them were not significant statistically. From the above, we could conclude that those herb medicines can be applied to patients effectively and further studies on effective fraction of some herbs are thought to be needed.

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Synergistic Effect of Panax ginseng and Cinnamoum Blume Mixture on the Inhibition of Cancer Cell Growth in vitro (인삼과 계피 혼합물에 의한 in vitro에서 암세포 증식억제의 상승 효과)

  • Chung Hwa Ryung;Lee Ji Young;Kim Dong Chung;Hwang Woo Ik
    • Journal of Ginseng Research
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    • v.23 no.2 s.54
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    • pp.99-104
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    • 1999
  • The effects of ginseng and cinnamon extract alone or mixture on the various cancer cell lines in vitro have been examined. Human colon cancer cell line (HT-29), human rectal cancer cell line (HRT-18) and human hepatoma cell line (HepG2) were used for the experiment. When given separately, the proliferation of all cancer cell lines was inhibited in proportion to the concentration of ginseng or cinnamon extract, respectively. Based on the cytotoxic activity, mixture of ginseng and cinnamon extract demonstrated a synergistic inhibition of cancer cell growth. The progression of cell cycle from G1 to S phase was significantly inhibited by ginseng and cinnamon mixture in the HT-29 and HRT-18 cell lines.

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Regulatory Role of Autophagy in Globular Adiponectin-Induced Apoptosis in Cancer Cells

  • Nepal, Saroj;Park, Pil-Hoon
    • Biomolecules & Therapeutics
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    • v.22 no.5
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    • pp.384-389
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    • 2014
  • Adiponectin, an adipokine predominantly secreted from adipose tissue, exhibits diverse biological responses, including metabolism of glucose and lipid, and apoptosis in cancer cells. Recently, adiponectin has been shown to modulate autophagy as well. While emerging evidence has demonstrated that autophagy plays a role in the modulation of proliferation and apoptosis of cancer cells, the role of autophagy in apoptosis of cancer cell caused by adiponectin has not been explored. In the present study, we demonstrated that globular adiponectin (gAcrp) induces both apoptosis and autophagy in human hepatoma cell line (HepG2 cells) and breast cancer cells (MCF-7), as evidenced by increase in caspase-3 activity, Bax, microtubule-associated protein light chain 3-II (LC3 II) protein levels, and autophagosome formation. Interestingly, gene silencing of LC3B, an autophagy marker, significantly enhanced gAcrp-induced apoptosis in both HepG2 and MCF-7 cell lines, whereas induction of autophagy by rapamycin, an mTOR inhibitor, significantly prevented gAcrp-induced apoptosis in hepatoma cells HepG2. Furthermore, modulation of autophagy produced similar effects on gAcrp-induced Bax expression in HepG2 cells. These results implicate that induction of autophagy plays a regulatory role in adiponectin-induced apoptosis of cancer cells, and thus inhibition of autophagy would be a novel promising target to enhance the efficiency of cancer cell apoptosis by adiponectin.

Effect of the Ethanol Extract of Vitis labrusca Root on Apoptosis in Hep G2 Cells (포도근 에탄올 추출물이 Hep G2 세포의 자연사에 미치는 효과)

  • Lee, Dong-Kyo;Lee, Kang-Pa;Kim, Hyuck;Choi, Byung-Jin;Chang, Hae-Ryong;Park, Won-Hwan
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.2
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    • pp.377-384
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    • 2008
  • The root of Vitis labrusca, is used as a source of health promoting drug in Korean traditional medicine. It has been reported that root of Vitis labrusca has antioxidant, anti lipid peroxidation and anti-reactive nitrogen species (RNS) activities. The aim of this study was to elucidate the molecular changes of apoptotic signaling pathways in phorbol 12-myristate 13 acetate (PMA)-induced human hepatocellular carcinoma cell line (Hep G2). The root of Vitis labrusca, ethanol extract (RVLEE) was tested for cell viability on Hep G2 cell using the MTT assay. RVLEE exhibited weak cytotoxic activity. However, treatment of Hep G2 cells with RVLEE suppressed PMA-induced cell proliferation. Also, dramatic changes of cell death signals in cellular molecules such as Chk2/Cds1, CIDE-B, CLIMP-63, Bax, Bcl-xL, C-myc, Bcl-2, Bric-5, NIP-3, TRAF2 and BAR but not CIDE-B and DR4. Futhermore, our results showed that the treatment of Hep G2 cells with 25 and $50\; {\mu}g/ml$ of RVLEE suppressed PMA-induced COX-2 gene activity. These data suggest that RVLEE have inhibitory effect of cell proliferation, induction of apoptosis and, thus, may offer therapeutic potential in Hep G2.

The effects of water extract from Dictamnus dasycarpus Turcz on Hepatocellular Damage in vitro (백선 추출물의 간세포 손상에 대한 연구)

  • Ha, Hun-Yong
    • The Korea Journal of Herbology
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    • v.29 no.5
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    • pp.91-95
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    • 2014
  • Objectives : This study was carried out to evaluate whether the water extract from cause the cellular damage in HepG2 cell line. It was reported that Dictamnus dasycarpus Turcz(DDT) intake induce poisoning symptoms in human population. These symptoms was closely related to liver toxicity, however, mechanisms for liver toxicity caused by DDT have not been elucidated exactly. Here, hepatotoxicity caused by DDT was evaluated using HepG2 cell line. Methods : Water extract of DDT was treated into HepG2 cell with various doses such as 0, 0.1, 0.5, 1.0 and $5.0mg/m{\ell}$. In order to cell viability, both MTT and LDH assay were carried out. Also, apoptosis array kit was used to identify whether cell death caused by DDT is due to apoptosis or not. In addition, reactive oxygen species (ROS) was measured after treatment of water extract. Results : We found out significant changes in the apoptosis related factors of hepatocyte. The cell viability of HepG2 treated with DDT water extract was decreased in dose-dependent. Also most of the apoptosis related factors were significantly increased. We found out that Caspase 3, Cytochrome C and ROS had increased in dose-dependent. In addition, other apoptosis related factors Bcl 2 and Bax, which were also constant changes. However, there was no significance. Conclusions : These results suggest that water soluble extract of DDT is expected to have oral toxicity, including hepatocellular damage Therefore, it is suggested that DDT could cause various side effects and toxicity of clinical conditions.