• 한국미생물·생명공학회지
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• 제49권4호
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• pp.594-602
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• 2021

The NLRP3 (nucleotide-binding domain, leucine-rich repeat family pyrin domain containing 3) inflammasome plays an important role in the initiation of inflammatory responses, through the recognition of pathogen-associated molecular patterns and tumor progression, including tumor growth and metastasis. In this study, we examined the effects of defective NLRP3 on the growth, migration, and invasiveness of hepatocellular carcinoma (HCC) SK-Hep1 cell. First, HCC SK-Hep1 cells were transfected with human NLRP3 targeting LentiCRISPRv2 vector using the CRISPR-Cas9 system, and NLRP3 deficiency was confirmed by RT-qPCR and western blotting. NLRP3 deficient SK-Hep1 cells showed delayed cell growth and decreased protein expression of PI3K, p-AKT, and pNF-κB when compared to NLRP3 complete SK-Hep1 cells. In addition, NLRP3 deficiency arrested the cell cycle at G1 phase through an increase in p21 and a reduction in CDK6. NLRP3 deficient SK-Hep1 cells also showed significantly delayed cell migration, invasion, and wound healing. The expression of epithelial-mesenchymal transition signaling molecules, such as N-cadherin and MMP-9, was found to be dramatically decreased in NLRP3 deficient SK-Hep1 cells compared to NLRP3 complete SK-Hep1 cells.

• 생약학회지
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• 제38권2호통권149호
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• pp.176-180
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• 2007

Five liglans, sauchinone (1), di-0-methyltetrahydrofuriguaiacin B (2), manassantin A (3), manassantin B (4) and saucerneol B (5), have been isolated from the MeOH extract of Saururus chinesis roots. The evaluation for protective effect of compounds 1-5 against tacrine-induced cytotoxicity in human liver-derived Hep G2 cells was conducted. Compounds 1,2, and 5 showed significant protective effects with the EC$_{50}$ values of74.2${\pm}$0.9, 111.3${\pm}$0.8,64.3${\pm}$0.8 ${\mu}$M, respectively. Silybin, one of the well-known hepatoprotective agents, used as a positive control, and also showed protective effect with an EC$_{50}$ value of 86.2${\pm}$0.5 ${\mu}$M.

• 약학회지
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• 제59권5호
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• pp.201-206
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• 2015

The aim of the present study was to optimize in vitro method for the prediction of drug-induced liver injury using human liver microsomes (HLM). Cytotoxicity test of cyclophosphamide and acetaminophen in HepG2 cells cultured with HLM showed that the newly established condition using 0.375 mg/ml HLM for 24 hr incubation was comparable or more sensitive than the previously established condition using 0.75 mg/ml HLM for 12 hr incubation. Although the cytotoxic effect of troglitazone was completely attenuated by 0.75 mg/ml HLM, it was augmented by 0.375 mg/ml HLM in the presence of the NADPH-generating system. The cytotoxic effect of chlormezanone, a withdrawn drug due to hepatotoxicity in human, was increased by HLM in the presence of the NADPH-generating system. In contrast, the cytotoxic effect of methapyrilene, a withdrawn drug due to hepatotoxicity in rats, was decreased by HLM in the presence of the NADPH-generating system. The present study suggests that the optimized in vitro method using HLM can be useful for the prediction of drug-induced hepatotoxicity.

• Journal of Ginseng Research
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• 제19권3호
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• pp.212-218
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• 1995

A human hepatoma cell line, hep G2, was used to investigate the mechanism of serum cholesterol reduction by ginseng total saponin, ginsenoside-$Rb_1$, - $Rb_2$, and non-saponin fraction (ether extraction). Hep G2 cells were incubated in 10 $\mu\textrm{g}$/ml of cholesterol containing serum free-RPMl1640 medium with various concentration of ginseng components. The amounts of cholesterol in Hep G2 cells were decreased to maximum 51% in total saponin or two ginsenoside-treated groups while there was 137% increase in cholesterol level of control group as compared with that of normal group. Nonsaponin groups did not show the same effect. In order to elucidate the observed changes in the amount of cholesterol, the activity of amyl CoA : cholesterol acyltransferase (ACAT) in groups showing remarkable reduction in cholesterol amount, i.e., total saponin 10-6%, ginsenoside-$Rb_1$ $10^{-4}$%, ginsenoside-$Rb_2$, $10^{-4}$%, and non-saponin fraction $10^{-4}$%, was assayed using [1-$^{-14}C$%]oleic acid as enzyme substrate. The activity of ACAT was increased in all groups tested as compared with that of control group except for non-saponin group cultured in water soluble cholesterol containing medium. The serum cholesterol lowering effects of ginseng components can partially be attributed to the increased hepatocellular ACAT activity.

• 생명과학회지
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• 제25권4호
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• pp.441-449
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• 2015

본 연구에서는 인체 폐암세포주인 A549 세포와 간암세포주인 HepG2 세포를 사용하여 Endlicheria anomala 메탄올 추출물(Methanol extract of E. anomala, MEEA)의 항암 활성 및 그 분자적 기전에 관하여 분석하였다. 먼저 MEEA가 인체 폐암세포와 간암세포의 증식에 미치는 영향을 분석한 결과, 암세포의 증식을 억제하는 효과가 탁월하였다. 그 후 MEEA에 의한 세포 증식이 억제되는 원인을 분석하기 위하여 Flow cytometry analysis, AnnexinV & 7-AAD 이중 염색을 수행한 결과, 두 세포에서 모두 농도의존적으로 apoptosis 유발군인 SubG1기의 세포 분포가 증가하였고, early apoptosis에서 late apoptosis로 전환되는 공통적인 현상을 확인할 수 있었다. 또한 apoptosis의 유발로 일어날 수 있는 세포의 형태 변화를 관찰하기 위한 DAPI 염색과 DNA fragmentation을 통하여 MEEA 처리에 의한 A549의 염색질 응축, 사멸체 형성 및 DNA의 끌림 현상을 관찰할 수 있었으며, 이와 관련된 분자적 기전 분석을 위한 Western blot을 추가로 수행하여 caspase, PARP, pro-, anti-apoptotic 단백질의 발현을 관찰하였다. 이상의 결과로 MEEA는 인체 폐암세포와 간암세포에서 p53의 발현 증가와 Bcl-2 family의 변화를 유도하며, caspase-3와 관련된 경로를 통하여 농도의존적으로 apoptosis를 유발시킨다는 것을 증명하였다. 이는 MEEA가 항암 활성을 보유하고 있고, 인체 폐암세포와 간암세포 사멸의 기전 연구를 위한 중요한 자료가 될 수 있음을 시사한다.

• 생명과학회지
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• 제10권1호
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• pp.79-85
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• 2000

Chemoprevention is one of the major strategies for cancer control. It is well established that dietary factors play an important role in modulating the development of certain types of human cancer. The experiment was conducted to determine quinone reductase(QR) activity induction of Daucus carota L. on HepG2 cells. Among various partition layers of roots of Daucus carota L., the ethyl acetate partition layer(DCMEA) and the n-hexane partition layer(DCMH) tested to be most effective which resulted 2.1 and 1.6 respectively compared to the control value of 1.0. In the case of seeds of Daucus carota L. n-butanol partition layer (DCMB) on HepG2 cells at a dose of 200 $\mu\textrm{g}$/$m\ell$ showed the highest induction activity of QR which was 3.0. These results suggest that potentially useful cancer chemoprevention chemicals could be isolated from DCMEA and DCMH of the roots and DCMB of the seeds of Daucus carota L.

• 생명과학회지
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• 제17권6호통권86호
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• pp.766-771
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• 2007

B형 간염 바이러스(HBV)와 C형 간염 바이러스(HCV)에 함께 감염되면 단독 감염의 경우보다 더 심각한 간질환이 유발되고 간암으로의 발전 가능성도 높아진다. 본 연구에서는 HBV의 X단백질(HBx)과 HCV의 코어 단백질이 인간 간암세포주인 HepG2세포에서 p53의 양을 협조적으로 증가시킨다는 것을 보여 주었다. 이로 인하여 세포예정사를 촉진하는 Bax 단백질의 발현이 더 증가하는 반면에 세포예정사를 억제하는 Bcl2의 발현은 더 억제됨이 관찰되었다. 그러나 이러한 효과들은 p53-음성인 Hep3B 세포에서는 관찰되지 않았다. 나아가 HBx와 코어 단백질은 HepG2의 cisplatin-매개성 세포예정사를 협조적으로 증가시키는 반면에 Hep3B에서는 이러한 효과가 나타나지 않았다. 이러한 연구 결과들은 HBV와 HCV가 동시에 감염되었을 경우에 나타나는 임상적인 소견을 이해하고 세포예정사에 미치는 HBx와 코어 단백질의 영향에 대한 기존의 상충적인 연구결과들을 해석하는데 도움을 줄 수 있다.

• 한국식품영양과학회지
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• 제30권1호
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• pp.86-91
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• 2001

The anticarcinogenic effects of various food components on human cancer cells have received much attention in recent years. The precise effect and mechanisms of anticarcinogens in food materials on cancer cells have rarely been investigated. This study was carried out to determine the effects of Daucus carota L. leaf (DCL) extracts on cytotoxic and chemopreventive effect on human cancer cells. The experiment was conducted to determine cytotoxicity of Daucus carota L. leaf extracts on HepG2, Hela and MCF-7 cells by MTT assay. Among various partition layers of Daucus carota L. leaf, the ethylacetate partition layer (DCLMEA) at 500 $\mu\textrm{g}$/mL was shown to be most effective on MCF-7 cell lines. The four partition layers which are DCLM, DCLMH, DCLMB and DCLMH were less effecitve in inducing cytotoxicity than DCLMEA was. We also determined the induction of intracellular quinone reductase (QR) activity by adding DCL extracts on HepG2 cells. Among various partition layers of DCL extracts, DCLMH and DCLM were tested to be most effective with results such as 4.9 and 4.73 with a control value of 1.0.

• 대한한의학방제학회지
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• 제15권2호
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• pp.127-137
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• 2007

The purpose of this study was to investigate the effect of Yong-dam-sa-gan-tang (YST) on apoptosis in HepG2 cells, First of all. to study the cytotoxic effect of methanol extract of YST on HepG2 cells, the cells were treated with various concentrations of YST and then cell viability was determined by XTT reduction method and trypan blue exclusion assay. YST reduced proliferation of HepG2 cells in a dose-dependent manner. To confirm the induction of apoptosis, HepG2 cells were treated with various concentrations of YST. The cleavage of poly AD P-ribose polymerase (P ARP), a substrate for caspase-3 and a typical sign of apoptosis, and the activation of caspase-3, procaspase-8 and procaspase-8 were examined by western blot analysis. YST decreased procaspase-3, procaspase-8 and procaspase-9 levels in a dose-dependent manner and induced the clevage of PARP. YST triggered the mitochondrial apoptotic signaling by increasing the release of cytochrome c from mitochondria to cytosol. Furthermore, YST also downregulated the anti-apoptotic Bcl-2 and upregulated the pro-apoptotic-Bax. Therefore, this result suggest that YST induced HepG2 cell death through the mitochondrial pathway. Sustained activation of the Ras/Raf/MEK/ERK cascade in cells results in a cell cycle arrest and has been implicated in the differentiation of certain cell types, in many cases acting to promote differentiation. YST decreased the activation of Ras/Raf/MEK/ERK cascade in a dose-dependent manner. These results suggest that YST is potentially useful as a chemo-therapeutic agent in HepG2.

• Biomolecules & Therapeutics
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• 제10권4호
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• pp.287-292
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• 2002

To find new inhibitory effects from oriental drugs, Albizziae root was extracted in methanol and the extracted was stepwisely fractionated by hexane, chloroform, ethylacetate, butanol and water. In cytotoxic effect of Albizziae root fractions against cancer cell lines including human hepatoma cells(HepG2) were investigated. Expecially the butanol fraction exhibited a inhibition effects on the growth of human hepatoma cells(HepG2). It inhibited of HepG2 cells with the value of IC50. The activities of qutathione after B(a)P treatment were markedly decreased than control, but those levels were increased by the treatment of Albizziae root methanol fraction. The activity of glutathione-S-transferase after B(a)P treatment were markedly decreased than control, but those levels were increased by the treatment of Albizziae root methanol traction. Induction of phase II enzymes is a major mechanism of chemoprevention. The induction levels of quinone reductase(QR) activity in cultured murine hepatoma(Hepa IcIc7)cell by methanol extract of Albizziae root were measured. Among the tested tractions, the extracts of butanol were found to induce QR activities over 2.8 fold than control. These results suggest that Albizziae root has chemopreventive Potential by inducing QR activities and GST levels and increasing GSH