• Archives of Pharmacal Research
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• 제27권4호
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• pp.402-406
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• 2004

This research team found in previous studies, that the ginseng saponin metabolite IH901 induces apoptosis in HepG2 cells via a mitochondrial-mediated pathway, which resulted in the activation of caspase-9 and subsequently of caspase-3 and -8. Based on these results, the involvement of the Fas/Fas ligand (FasL) death-receptor pathway, in IH901-induced apoptosis in HepG2 cells, was investigated. Levels of Fas and the Fas ligand (FasL) mRNA or protein were not increased by IH901, rather they were decreased significantly at 18 h post treatment. Soluble FasL (sFasL) was detectable by immunoprecipitation analysis En the medium of HepG2 cells treated with IH901. Increased levels of sFasL were inversely correlated with the levels of FasL. Preincubation of HepG2 cells with antagonistic anti-Fas antibody showed little protective effect, if any, on IH901-induced cell death. At a $30{\mu}M$ (24 and 48 h) and $40{\mu}M$ (24 h) concentration of IH901, the cytotoxic effect of IH901 was less then $50\%$, anti-Fas antibody prevented IH901-induced cell death. However, at a $60{\mu}M$ (24 and 48 h) and $40{\mu}M$ (48 h) concentration of IH901, cell death rates were about $80\%$ or more and most of the chemopreventive and chemotherapeutic effects of IH901 were manifested. Blocking the Fas receptor did not influence IH901-induced cell death. These results indicate that the Fas/FasL system is engaged, but not required for IH901-induced cell death, at pharmacologically significant concentrations.

• 대한한의학회지
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• 제39권4호
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• pp.158-170
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• 2018

Objectives: The objective of this study is Korean mistletoe lectin (Viscum album coloratum agglutinin, VCA) and bee venom (BV) to experimental prove comparative study of VCA and BV on the anti-cancer effect and mechanisms of action. Methods: In this study, it was examined in a human hepatocellular carcinoma cell line, Hep G2 cells. Cytotoxic effects of VCA and BV on Hep G2 cells were determined by 3- (4, 5-dimethylthiazol-2-yl) -2, 5-diphenyltetrazolium bromide (MTT) assay in vitro. VCA and BV killed Hep G2 cells in a time- and dose-dependent manner. Results: The apoptotic cell death was then confirmed by propidium iodide staining and DNA fragmentation analysis. The mechanisms of action was examined by the expression of anti-apoptotic proteins and activation of mitogen-activated protein kinases. Treatment of Hep G2 cells with VCA activated poly (ADP-ribose) polymerase-1 (PARP-1) known as a marker of apoptosis, and mitogen-activated protein kinases signaling pathways including SAPK/JNK, MAPK and p38. BV also activated PARP-1, MAPK, p38 but not JNK. The expression level of anti-apoptotic molecule, Bcl-X, was decreased by VCA treatment but not BV. Finally, the phosphorylation level of ERM proteins involved in the cytoskeleton homeostasis was decreased by both stimuli. Conclusion: We examined the involvement of kinase in VCA or BV - induced apoptosis by using kinase inhibitors. VCA-induced apoptosis was partially inhibited by in the presence.

• Asian Pacific Journal of Cancer Prevention
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• 제16권15호
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• pp.6417-6421
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• 2015

Viscum album var (VAV) also known as mistletoe, has long been categorized as a traditional herbal medicine in Asia. In addition to its immunomodulating activities, mistletoe has also been used in the treatment of chronic hepatic disorders in China and Korea. There are numerous reports showing that VAV possesses anti-cancer effects, however influence on human hepatocarcinoma has never been elucidated. In the present study, hot water extracts of VAV was evaluated for its potential anti-cancer effect in vitro. SK-Hep1 cells were treated with VAV (50-400ug/ml) for both 24 and 48 hours then cell viability was measured by cell counting kit-8 (CCK-8). Flow cytometry analysis was used to measure the proportion of SK-Hep1 in the different stages of cell cycle. RT-PCR and Western blot analysis were conducted to measure expression of cell cycle arrest related genes and proteins respectively. VAV dose dependently inhibited the proliferation of SK-Hep1 cells without any cytotoxicity with normal Chang liver cell (CCL-13). Flow cytometry analysis showed that VAV extract inhibited the cell cycle of SK-Hep1 cells via G1 phase arrest. RT-PCR and Western blot analysis both revealed that cyclin dependent kinase 2 (Cdk2) and cyclin D1 gene expression were significantly down regulated while p21 was upregulated dose dependently by VAV treatment. Combined down regulation of Cdk2, Cyclin D1 and up regulation of p21 can result in cell death. These results indicate that VAV showed evidence of anti-cancer activity through G1 phase cell cycle arrest in SK-Hep1 cells.

• 약학회지
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• 제56권2호
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• pp.80-85
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• 2012

Previously, we have reported that arachidonic acid (AA) appears to be involved in the induction of apoptosis in HepG2 human hepatoblastoma cells. In this study we investigated the possible role of the NADPH oxidase, a membranebound enzyme generating reactive oxygen species (ROS), in the mechanism of AA-induced apoptosis in HepG2 cells. Apoptotic cell death induced by AA was significantly suppressed by various inhibitors of the NADPH oxidase, diphenylene iodonium (DPI), apocynin (Apo) and neopterine (NP). In addition, these inhibitors of the NADPH oxidase completely blunted the AA-induced ROS elevation. Next, we investigated the implication of metabolic pathway of AA in these AA actions. Both apoptosis and ROS production induced by AA were not significantly altered by treatment with indomethacin (Indo) or nordihydroguaiaretic acid (NDGA), selective inhibitors of cyclooxygenase (COX) and lipoxygenase (LOX), respectively, suggesting that AA metabolites produced by COX or LOX may not have an essential role in the AA-induced apoptosis and ROS generation. Collectively, these results suggest that the NADPH oxidase may be a key player in the mechanism of AA-induced apoptosis in HepG2 cells. These results further suggest that NADPH oxidase may be a good target for the management of human hepatomas.

• Asian Pacific Journal of Cancer Prevention
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• 제15권4호
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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.

• 생약학회지
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• 제48권4호
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• pp.273-279
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• 2017

The aim of this study was to investigate the mechanisms underlying apoptosis induced by a broussochalcone B (BCB) from Broussonetia papyrifera in HepG2 cells. The results showed that BCB treatment for 24 hr significantly inhibited cell viability in a dose-dependent manner, and induced apoptosis in HepG2 cells. More so, BCB treatment triggered the cleavage of caspase-8, -9, -3, poly (ADP-ribose) polymerase (PARP), increase of Bax level, and decrease of Bcl-2 expression. A general caspase inhibitor (z-VAD-fmk) blocked BCB-induced cell death. Furthermore, BCB treatment caused reactive oxygen species (ROS) production in a dose-dependent manner. In addition, an antioxidant N-acetylcysteine (NAC) blocked BCB-induced ROS production and cell death. Therefore, these results indicate that BCB-induced apoptosis is mediated by a caspase dependent pathway and ROS production in HepG2 cells.

• 대한한방내과학회지
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• 제25권1호
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• pp.81-91
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• 2004

Objectives : The aim of this study is to investigate the inhibitory effect of Chungganhaeju-Tang on alcohol induced human hepatic cell apoptosis by synthesis of glutathione. Methods : The amount of glutathione in HepG2 cell was measured with colorimetric glutathione assay kit and glutathione-conjugated CDNB(1-chloro-2,4-dinitrobenzene) at $37^{\circ}C$ and then measured by spectrometry to assess the activity of glutathione S-transferase. Results : The synthesis of glutathione and the activity of glutathione S-transferase in HepG2 cell were promoted by Chungganhaeju-Tang and increased in dose/time-dependent manner. Chungganhaeju-Tang inhibited apoptosis induced by ethanol and acetaldehyde dependent to treatment dosage. In Buthione sulfoximine, a glutathione synthesis inhibitor, treated case, the synthesis of glutathione was inhibited and in Chungganhaeju-Tang treated case, the synthesis of glutathione is promoted with or without Buthione sulfoximine. The present findings suggest that Chungganhaeju-Tang inhibits alcohol induced apoptosis by synthesis of glutathione in HepG2 cell. Conclusions : The result indicates that Chungganhaeju-Tang protects human hepatic cell by glutathione synthesis and made the liver recover from alcohol induced damage.

• 생명과학회지
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• 제28권1호
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• pp.50-57
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• 2018

Millettia erythrocalyx는 콩과(Fabaceae)에 속하는 식물로 중국, 태국, 인도 등 열대 아열대 지역에 분포하며, 항바이러스 활성을 보유하고 있다는 보고가 있으나 항산화능과 항암활성 등에 관한 연구는 보고된 바가 없다. 따라서 본 연구에서는 M. erythrocalyx의 에탄올 추출물(EEME)을 사용하여 항산화능을 측정하고, 인체간암세포주인 HepG2에 대한 항암활성과 그 분자적 기전에 관하여 분석하였다. 먼저 DPPH radical scavenging activity를 통해 분석한 결과, EEME의 $IC_{50}$는 $2.74{\mu}g/ml$로 뛰어난 항산화능을 보유하였음을 확인하였다. 또한 EEME 농도 의존적으로 HepG2 세포의 성장을 억제하였다. EEME의 HepG2 세포 사멸 효과의 기전을 분석하기 위하여 세포주기를 분석한 결과, EEME 농도의존적으로 SubG1 세포가 증가하였으며, Annexin V 염색과 DAPI 염색을 통해 apoptotic 세포 및 apoptotic body가 증가됨을 확인하였다. 또한 apoptosis 관련 단백질들의 발현변화를 분석한 결과, EEME 처리에 의해 사멸수용체인 Fas와 pro-apoptotic 단백질인 Bax의 발현이 증가되었으며, caspase-3, -8, -9가 활성화되고 최종적으로 PARP가 분해되어 apoptosis가 유도되었음을 확인하였다. 이러한 결과들로부터 EEME는 내인성 및 외인성 경로를 통한 apoptosis 유도에 의하여 HepG2 세포의 증식을 억제하는 항암활성을 보유하였음을 확인하였다.

• 생명과학회지
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• 제28권2호
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• pp.176-186
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• 2018

김(Porphyra yezoensis, Laver)의 MeOH 추출에 의한 유기용매 별 분획물에서 폴리페놀 함량과 항산화 활성 및 간암세포주 HepG2의 세포증식 억제효과를 확인하였다. $CHCl_3$ 분획물의 폴리페놀 함량은 $10.34{\mu}g/mg$으로 물 분획물의 $13.08{\mu}g/mg$ 보다는 다소 적게 나타났으나, DPPH 자유라디칼 소거에 의한 전자공여능(EDA)에서 나타난 $ED_{50}$은 $16.96{\mu}g/ml$로 가장 높게 나타났다. $CHCl_3$과 EtOAc 분획물은 농도의존적으로 HepG2 세포의 증식을 억제하였으며, 특히 $900{\mu}g/ml$의 $CHCl_3$ 분획물을 24시간 동안 처리하여 90%의 세포증식이 억제되었다. 한편 $CHCl_3$ 분획물이 처리된 HepG2 세포의 유전자 발현 양상을 microarray로 확인하였다. P. yezoensis의 효능과 연관지은 gene ontology 분석으로 비타민 D 합성 과정, 항균작용에 대한 반응 및 영양물질에 대한 반응에 관련된 유의 유전자들을 탐색하였다. 유의 유전자로 IL6R와 CYP1A1를 선정하였고, 이들 유전자의 상위 조절자는 ARNT 유전자가 선정되었다. 또한 50 및 $100{\mu}g/ml$의 $CHCl_3$ 분획물이 처리된 HepG2 세포에서 IL6R와 CYP1A1 단백질의 발현과 상위 조절자인 ARNT의 활성을 Western blotting으로 확인하였다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권21호
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• pp.9319-9325
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• 2014

Alkaloids are the most extensively featured compounds of natural anti-tumor herbs, which have attracted much attention in pharmaceutical research. In our previous studies, a mixture of major three alkaloid components (5, 6-dihydrobicolorine, 7-deoxy-trans-dihydronarciclasine, littoraline) from Hymenocallis littoralis were extracted, analyzed and designated as AHL. In this paper, AHL extracts were added to human liver hepatocellular cells HepG-2, human gastric cancer cell SGC-7901, human breast adenocarcinoma cell MCF-7 and human umbilical vein endothelial cell EVC-304, to screen one or more AHL-sensitive tumor cell. Among these cells, HepG-2 was the most sensitive to AHL treatment, a very low dose ($0.8{\mu}g/ml$) significantly inhibiting proliferation. The non-tumor cell EVC-304, however, was not apparently affected. Effect of AHL on HepG-2 cells was then explored. We found that the AHL could cause HepG-2 cycle arrest at G2/M checkpoint, induce apoptosis, and interrupt polymerization of microtubules. In addition, expression of two cell cycle-regulated proteins, CyclinB1 and CDK1, was up-regulated upon AHL treatment. Up-regulation of the Fas, Fas ligand, Caspase-8 and Caspase-3 was observed as well, which might imply roles for the Fas/FsaL signaling pathway in the AHL-induced apoptosis of HepG-2 cells.