• 한국식품과학회지
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• 제36권4호
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• pp.662-668
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• 2004

13종의 한국산 야생버섯 추출물의 항돌연변이 효과의 검토는 S. typhimurium TA98과 TA100을 이용한 Ames 실험계로 확인하였다. 그 결과, 직접변이원인 MNNG에 대해 산속그물버섯 아재비와 구름버섯의 메탄올 추출물은 TA100에서 90-94%의 강한 항변이원성을 보였고, 갓버섯과 구름버섯의 열수 추출물도 58-61%의 저해효과를 보였다. 간접변이원인 B(a)P에 대해서는 TA98과 TA100에서 구름버섯과 턱받이금버섯 메탄올 추출물이 가장 강한 저해효과를 나타내었고, 열수 추출물중 잔피막흑주름버섯과 수원무당버섯은 TA98에서 60-70% 정도의 저해활성을, 갓버섯은 TA100에서 45% 정도의 돌연변이 억제효과를 볼 수 있었고, 나머지 버섯시료는 저해활성을 거의 보이지 않았다. 한편 야생버섯 메탄올 추출물 0.5mg/mL 투여시 결장암세포인 HT29에 대해 그물버섯, 산속그물버섯아재비, 방패외대버섯, 황소비단그물버섯, 구름버섯, 턱받이금버섯, 덕다리버섯에서 55-78% 이상의 억제 효과를 보였고, 1 mg/mL의 시료농도에서는 11종의 버섯에서 50% 이상의 억제 효과를 나타내었다. 간암세포인 HepG2에서 대해서는 방패외대버섯 메탄올 추출물 1 mg/mL에서만 58.4%의 저해능을 보였고, 나머지 시료에서는 45% 정도의 낮은 저해활성을 보였다. 혈액암세포주인 U937의 경우도 그물버섯, 산속그물버섯아재비, 수원무당버섯, 황소비단 그물버섯, 밀버섯, 잔피막흑주름버섯 메탄올 추출물에서 60% 이상의 높은 저해 활성을 나타내었다. 구름버섯 열수추출물의 경우 HepG2와 U937에 대해서 53-65% 정도의 비교적 높은 저해활성을 보였고, 대부분의 버섯 열수 추출물은 40%이하의 낮은 저해활성을 보였다. 정상 임파구에 대해서 야생버섯 추출물 0.5-1 mg/mL 농도에서 75-80% 이상의 생존율을 보여, 야생버섯 추출물을 1 mg/mL 농도로 처리할 경우 암세포에 대한 높은 억제효과에 비해 정상세포에 대해서는 비교적 낮은 독성을 가짐을 확인하였다.

• Journal of Microbiology and Biotechnology
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• 제12권3호
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• pp.470-475
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• 2002

In the course of screening for a novel cell cycle inhibitor, a potent Cdk 1 inhibitor, HY558, was found from the culture broth of Penicillium minioluteum F558 isolated from a soil sample. The molecular ion of HY558 was identified at m/z 329 (MH+) with a molecular formula of $C_20H_44ON_2$. HY558 exhibited selective antiproliferative effects on various human cancer cell lines. Its $IC_50$ values were estimated to be 0.29 mM on HepG2, 0.30 mM on HeLa, 0.30 mM on HL6O, 0.33 mM on HT-29, and 0.25 mM on AGS cells. Interestingly, Hy558 demonstrated no antiproliferative effect with normal lymphocytes used as the control, and a low level of inhibition on the proliferation of A549 cancer cells. A flow cytometric analysis of HepG2 cells revealed an appreciable arrest of cells at the G1 and G2/M phases of the cell cycle following treatment with Hy558. furthermore, DNA fragmentation due to apoptosis was observed in HeLa cells treated with 0.46 mM of HY558.

• 한국식품영양과학회지
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• 제34권6호
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• pp.771-775
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• 2005

본 실험은 홍조해조류의 하나인 불등가사리를 추출 후 각 용매별로 분획하여 암세포 성장억제 효과와 QR유도활성 효과 등 생리활성을 연구하였다. 불등가사리를 이용하여 4종의 인체 암세포주HepG2, HeLa, MCF-7및 HT-29와 정상 간세포에 대한 암세포 증식억제 실험을 한 결과 사용한 4종의 암세포주에서 모두 시료첨가 농도에 의존적으로 증식 저지 효과가 나타났고, 특히 불등가사리의 methanol 분획층인 GFMM에서는 매우 낮은 농도의 시료첨가에도 불구하고 괄목할 만한 높은 암세포 성장 억제효과를 나타내었으며, 정상 간세포에서는 모든 분획층에서 약한 세포증식 억제율을 나타내어 정상 세포에 미치는 독성이 낮음을 확인하였다. 특히 본 시료는 여성암의 대표적인 유방암과 자궁경부암의 암세포 성장저지효과가 탁월하였다. 한편, 사용한 4가지 암세포주 중 유일하게 QR을 가지고 있는 HepG2를 이용한 암 예방지표인 QR 효소 유도 활성 여부를 측정한 결과 분획물 첨가농도를 10, 20, 30 및 40 $\mu$g/mL로 첨가하였을 때 GFMM의 첨가농도 $20\mu g/mL$에서 대조군에 비해 약 2배 이상의 높은 QR 유도효과를 나타내어 최종농도 $40\mu g/mL$에서는 2.6의 암 예방 QR 유도효과를 나타내었다.

• Archives of Pharmacal Research
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• 제27권9호
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• pp.947-953
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• 2004

The antioxidative and hepatoprotective potentials of two anthraquinones, alaternin (2-hydroxy-emodin) and emodin, to scavenge and/or inhibit hydroxyl radicals generated by the Fenton reaction and to protect tacrine-induced cytotoxicity in human liver derived HepG2 cells were evaluated, respectively. The inhibitory activity on hydroxyl radical generated in a cell-free chemical system (FeSO$_4$/$H_2O$$_2$) was investigated by a fluorescence spectrophotometer using a highly fluorescent probe, 2$^1$,7$^1$-dichlorofluorescein. The hydroxyl radical scavenging activity was determined by electron spin resonance spectroscopy using 5,5-dimethy-1-pyrroline-N-oxide as hydroxyl radicals trapping agents. Tacrine-induced HepG2 cell toxicity was determined by a 3-［4,5-dimethylthiazole-2yl］-2,5-diphenyltertrazolium bromide assay. Although the scavenging activity of alaternin on hydroxyl radical was similar to that of emodin in dose-dependent pat-terns, the inhibitory activity exhibited by the former on hydroxyl radical generation was stron-ger than that of the latter, with $IC_{50}$/ values of 3.05$\pm$0.26 $\mu$M and 13.29$\pm$3.20 $\mu$M, respectively. In addition, the two anthraquinones, alaternin and emodin showed their hepatoprotective activ-ities on tacrine-induced cytotoxicity, and the EC$_{50}$ values were 4.02 11M and 2.37 $\mu$M, respec-tively. Silymarin, an antihepatotoxic agent used as a positive control exhibited the EC$_{50}$ value of 2.00 $\mu$M. These results demonstrated that both alaternin and emodin had the simultaneous antioxidant and hepatoprotective activities.ies.

• 생명과학회지
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• 제25권1호
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• pp.44-52
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• 2015

본 연구에서는, 암세포 증식 억제효능과 항산화, 항염증 효능을 동시에 가지는물질을 탐색하였다. 그 결과, 광곽향 메탄올 추출물이 A549, HepG2, MCF7, HT29 등 다양한 암세포에 대하여 세포성장억제효과를 보였고, A549에 대해 특이적으로 뛰어난 사멸효과를 보였다. 광곽향 메탄올 추출물에 의한 A549에서의 항암 효과는 p38 - Cdc25A - Cdk - Cyclin - Rb pathway를 통해 G1 arrest 유도로 연결되는 것으로 사료된다. 또한 DPPH를 통한 free radical의 소거능 확인 결과 항산화 효과를 가지고 있는 것을 확인하였고, 대식세포(RAW 264.7)의 iNOS 발현을 감소시켜 LPS에 의해 유도되는 NO의 생성을 유의적으로 억제함을 확인했다. 이러한 결과들로부터 광곽향 메탄올 추출물이 항산화, 항염증, 항암 후보물질 소재로 활용가능 할 뿐 아니라, 다양한 건강기능성 소재로 활용가능할 것이라 사료된다.

• Asian Pacific Journal of Cancer Prevention
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• 제14권9호
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• pp.5397-5402
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• 2013

The rates of morbidity and mortality of hepatocellular carcinoma (HCC) have not lessened because of difficulty in treating tumor metastasis. Mongolian Saussurea involucrata (SIE) possesses various anticancer activities, including apoptosis and cell cycle arrest. However, detailed effects and molecular mechanisms of SIE on metastasis are unclear. Thus, the present study was undertaken to investigate antimetastatic effects on HCC cells as well as possible mechanisms. Effects of SIE on the growth, adhesion, migration, aggregation and invasion of the SK-Hep1 human HCC cell line were investigated. SIE inhibited cell growth of metastatic cells in dose- and time-dependent manners. Incubation of SK-Hep1 cells with $200-400{\mu}g/mL$ of SIE significantly inhibited cell adhesion to gelatin-coated substrate. In the migration (wound healing) and aggregation assays, SIE treated cells showed lower levels than untreated cells. Invasion assays revealed that SIE treatment inhibited cell invasion capacity of HCC cells substantially. Quantitative real time PCR showed inhibitory effects of SIE on MMP-2/-9 and MT1-MMP mRNA levels, and stimulatory effects on TIMP-1, an inhibitor of MMPs. The present study not only demonstrated that invasion and motility of cancer cells were inhibited by SIE, but also indicated that such effects were likely associated with the decrease in MMP-2/-9 expression of SK-Hep1 cells. From these results, it was suggested that SIE could be used as potential anti-tumor agent.

• Journal of Microbiology
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• 제37권2호
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• pp.86-89
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• 1999

An OTHBVS cell line from HepG2 was established. This cell line stably expresses the human hepatitis B virus (HBV) middle S protein that includes the preS2 region which is important for HBV particle entry into the hepatocyte. To establish this cell line, the middle S open reading frame (ORF), with a promoter located in the 5' region and enhancer located in the 3' region, was cloned downstream from the metallothionine (MT) promoter of the OT1529 vector. In this vector, expression of the middle S protein was constructed to be regulated by its own promoter and enhancer. Expression of the large S protein which contains the preS1 region in addition to the middle S protein was designed to be regulated by the MT promoter. When extracts of OTHBVS cells were examined with an S protein detection kit (RPHA, Korea Green Cross Co.), an S protein was detected. Total mRNA of OTHBVS cell examined by northern blot analysis with an S ORF probe revealed small/middle S transcripts (2.1 kb). When the MT promoter was induced by Zn, large S transcripts (2.4 kb) were detected. The GP36 and GP33 middle S proteins were presumably detected, but large S proteins were not detected by immunostain analysis using anti-preS2 antibody.

• 한국식품과학회지
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• 제52권5호
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• pp.476-481
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• 2020

본 연구에서는 간보호 활성이 우수한 것으로 알려진 단풍취의 활용가치를 높이고 향후 건강기능성식품 소재로써의 가능성을 알아보고자 LPS/D-GalN 독성유발에 따른 단풍취 열수 추출물의 간기능 개선효과를 연구하였다. 단풍취 열수 추출물 내에 존재하는 5종의 Caffeoylquinic acid (CQA) 분석결과 4,5-di-O-caffeoylquinic acid (4,5-DCQA) 11.16 mg/g, 3,4-di-O-caffeoylquinic acid (3,4-DCQA) 5.23 mg/g, 5-O-caffeoylquinic acid (5-CQA) 4.88 mg/g, 3,5-di-O-caffeoylquinic acid (3,5-DCQA) 3.51 mg/g 및 4-O-caffeoylquinic acid (4-CQA) 3.31 mg/g 순으로 나타났다. 단풍취 열수 추출물이 함유하고 있는 총 폴리페놀 함량은 74.03 mg/g이었으며, ABTS cation radical 소거능은 추출물의 농도 50, 100, 200 및 400 ㎍/mL의 범위에서 각각 12.2, 25.0, 40.8, 68.55%를 나타냈고 DPPH radical 소거능은 50 ㎍/mL의 농도에서 54.38%를 나타냈다. HepG2 세포에 LPS/D-GalN을 처리하여 간독성을 유발한 결과 정상군을 제외한 모든 처리군에서 GGT, AST 및 LDH 활성이 증가하였고, 단풍취 열수 추출물 300 mg/mL을 처리하였을 때 대조군과 비교하여 GGT, AST 및 LDH 활성이 유의적으로 감소하는 것으로 나타났다. LPS/D-GalN을 처리하였을 때 정상군과 비교하여 모든 처리군에서 TNF-α 분비가 유의적으로 증가하는 것을 확인 할 수 있었으며, 단풍취 열수 추출물을 100 ㎍/mL 이상 처리하였을 때 TNF-α의 분비가 유의하게 감소하였다. 이상의 결과로 볼 때 단풍취 열수 추출물은 항산화 활성 증가 및 GGT, AST, LDH의 활성을 감소시키고 TNF-α 분비를 억제시킴으로써 LPS/D-GalN으로 인한 간 손상을 예방하는 것으로 판단된다.

• 한국식품영양과학회지
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• 제30권1호
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• pp.143-151
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• 2001

A marine bacterium producing carotenoid was isolated from the Yosu coastal area of South Korea, which was recorded as MCK-1. It was identified as Erythrobacter sp. Optimium conditions of marine carotenoid fermentation from Erythrobacter sp. were pH 6.0, a temperature of $25^{\circ}C$, 16 mM mannitol as a carbon source, 0.5% tryptone as a nitrogen source, 0.1 mM $Fe^{+2}$ ion as a mineral source and 1$\mu$M of cyanocobalamine as a growth factor in a jar-fermentor. Erythrobacter sp. was produced 351.27 mg/100mL of the marine carotenoid in these optimum conditions. This marine carotenoid was composed of 4 different conpounds, like as notoxanthin (61.4%), can thaxanthin (24.6%), fucoxanthin (8.2%), and zeaxanthin (5.8%). Physiological properties including antibacterial activity, cytotoxic effect, antioxidative effect and free radical scavenging activity were characterized with crude carotenoid. Carotenoid exhibited no antibacterial activity against E. coli and lactobacillus bulgaricus, but showed cytotoxic effect against cancer cells such as HepG2 (Hepatocellular carcinoma, human, ATCC HB-8065) and HeLa (Cervical carcinoma, human, ATCC CCL-2) cells. The impediment ratios for HepG2 and HeLa cell were 37.14% and 33.78%, respectively. This carotenoid expressed a strong antioxidative effect (77%) against CCL-13 5 $\mu\textrm{g}$/mL and 50 $\mu\textrm{g}$/mL crude carotenoid, respectively.

• Asian Pacific Journal of Cancer Prevention
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• 제16권15호
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• pp.6581-6589
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• 2015

Background: Previously, stingless bee (Trigona spp.) products from East Kalimantan, Indonesia, were successfully screened for in vitro antiproliferative activity against human cancer derived cell lines. It was established that propolis from T. incisa presented the highest in vitro cytotoxicity against the SW620 colon cancer cell line (6% cell survival in $20{\mu}g/mL$). Materials and Methods: Propolis from T. incisa was extracted with methanol and further partitioned with n-hexane, ethyl acetate and methanol. The in vitro cytotoxicity of the extracts was assessed by the MTT assay against human colon (SW620), liver (Hep-G2), gastric (KATO-III), lung (Chago) and breast (BT474) cancer derived cell lines. The active fractions were further enriched by silica gel quick column, absorption and size exclusion chromatography. The purity of each fraction was checked by thin layer chromatography. Cytotoxicity in BT-474 cells induced by cardanol compared to doxorubicin were evaluated by MTT assay, induction of cell cycle arrest and cell death by flow cytometric analysis of propidium iodide and annexin-V stained cells. Results: A cardol isomer was found to be the major compound in one active fraction (F45) of T. incisa propolis, with a cytotoxicity against the SW620 ($IC_{50}$ of $4.51{\pm}0.76{\mu}g/mL$), KATO-III (IC50 of $6.06{\pm}0.39{\mu}g/mL$), Hep-G2 ($IC_{50}$ of $0.71{\pm}0.22{\mu}g/mL$), Chago I ($IC_{50}$ of $0.81{\pm}0.18{\mu}g/mL$) and BT474 (IC50 of $4.28{\pm}0.14{\mu}g/mL$) cell lines. Early apoptosis (programmed cell death) of SW620 cells was induced by the cardol containing F45 fraction at the $IC_{50}$ and $IC_{80}$ concentrations, respectively, within 2-6 h of incubation. In addition, the F45 fraction induced cell cycle arrest at the G1 subphase. Conclusions: Indonesian stingless bee (T. incisa) propolis had moderately potent in vitro anticancer activity on human cancer derived cell lines. Cardol or 5-pentadecyl resorcinol was identified as a major active compound and induced apoptosis in SW620 cells in an early period (${\leq}6h$) and cell cycle arrest at the G1 subphase. Thus, cardol is a potential candidate for cancer chemotherapy.