• KSBB Journal
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• 제28권5호
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• pp.332-337
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• 2013

호열성 진정세균인 Thermotoga maritima의 xylose isomerase 유전자를 대장균을 이용하여 클로닝하고 재조합 발현시켰다. 재조합 효소의 최적활성은 $90^{\circ}C$와 pH 8.0에서 관찰되었다. 사포닌을 재조합효소로 처리한 후 사람의 위암 세포주 (AGS)와 대장암 세포주 (HT-29)에 처리한 결과, 효소 무처리 사포닌에 비해 우수한 암세포 생육저해 활성을 나타냈다. 한편, 푸코이단을 재조합효소로 처리한 후 동일 세포주들에 처리한 결과, 효소 무처리 푸코이단과 비슷한 암세포 생육저해 활성을 보였다. 1 ${\mu}g/ml$ 농도의 효소 처리 사포닌은 100 ${\mu}g/ml$ 농도의 효소 무처리 사포닌과 유사하거나 우수한 암세포 생육저해 활성을 보였다. 본 연구결과는 기능성 식품이나 의약품의 개발에 참고가 될 것으로 사료된다.

• 한국환경성돌연변이발암원학회:학술대회논문집
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• 한국환경성돌연변이발암원학회 2001년도 Korean Environmental Mutagen Society
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• pp.185-185
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• 2001

• 한국식품영양과학회지
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• 제29권4호
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• pp.726-731
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• 2000

본 연구에서는 단 삼(Salvia miltiorrhiza)의 추출물이 인체 결장암세포인 HT-29, 간암세포인 HepG2 및 직장 암세포인 HRT-18의 증식에 미치는 영향을 in vitro에서 확인하였다. 암세포의 배양액에서 단삼의 수용성 추출물과 에탄올 추출물의 암세포 증식억제효과는 농도에 비례하여 에탄올 추출물에서 더 효과적이었다. 이를 토대로 단삼의 에탄올 추출물의 급성독성, 수명연장 및 고형암형 성억제를 살펴보기 위해 in vivo 실험을 하였다. 급성독성실험결과 대조군의 15일 째의 평균체중은 32.3 g 이었고 실험군은 31.6 g으로 정상적인 상태를 유지하였으며, 흰쥐의 육종암세포인 sarcoma-180를 접종한 mouse의 수명연장실험결과 대조군에 비해 61%의 수 명연장 효과가 있음을 관찰하였다. 고형암억제 실험 결과에서도 대조군에 비해 에찬올 추출 물 처리군이 35%의 고형암 형성억제능을 나타내었다. 따라서 단삼의 에탄올 추출물 중에는 in vitro와 in vivo 실험을 통해 항암활성을 갖는 성분이 존재하며, 이성분은 유효한 항암제 로 개발될 수 있으리라 사료된다.

• 한국식품저장유통학회지
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• 제21권6호
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• pp.878-884
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• 2014

본 연구에서 고추장 제조시 발효가 항돌연변이 효과와 in vitro 항암효과에 미치는 영향을 확인하기 위하여 고추장 제조 재료인 원료 밀, 1차발효밀, 2차발효밀, 최종발효밀, 고춧가루와 최종발효밀로 제조된 고추장의 Sal. Typhimurium TA100을 이용한 Ames test를 실시하여 MNNG에 대한 항돌연변이 효과를 살펴보았으며, HT-29 인체 대장암세포와 AGS 인체 위암세포의 성장 억제효과를 살펴보았다. 실험한 결과 원료 밀보다 1차발효밀, 2차발효밀보다 최종발효밀이 발효진행에 따라 점차 증진된 항돌연변이 효과와 in vitro 항암효과가 나타났으며 최종발효밀과 고춧가루로 제조된 고추장이 가장 높았다. 15, 30일 발효시킨 고추장이 발효되지 않은 고추장보다 더 높은 항암효과가 나타났다. 따라서 발효과정(기간)이 고추장의 항돌연변이 효과와 항암효과에 중요하게 관련되어 있다고 할 수 있으며 고추장의 주재료인 고춧가루의 capsaicin, 비타민C와 ${\beta}$-carotene 등의 작용만이 아닌 밀에서 온 발효 숙성 중에 많이 증식된 미생물 그리고 발효에 의한 생성된 다른 활성물질이 항돌연변이와 항암 효과에 관련되어 있다고 하겠다.

• Journal of Microbiology and Biotechnology
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• 제19권11호
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• pp.1482-1489
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• 2009

In this study, we investigated the anti-proliferative effect of polysaccharides from Salicornia herbacea on HT-29 human colon cancer cells. Crude polysaccharides from S. herbacea (CS) were prepared by extraction with hot steam water, and fine polysaccharides from S. herbacea (PS) were obtained through further size exclusion chromatography. The anti-proliferative effect of CS and PS were measured using the MTS assay, apoptosis analysis, cell cycle analysis, and RT-PCR. HT-29 cells were treated with CS or PS at different dosages (0.5, 1, 2, 4 mg $ml^{-1}$) for 24 or 48 h. CS and PS inhibited proliferation and stimulated apoptosis of cells in a dose-dependent manner. Flow cytometric analysis after Annexin V-FITC and PI staining revealed that treatment with CS or PS increased total apoptotic death of cells to 24.99% or 91.59%, respectively, in comparison with the control (13.51 %). PS increased early apoptotic death substantially - up to 12 times more than the control. Treatment with CS or PS resulted in a concentration-dependent increase of the G2/M cell population of the cell cycle as determined by flow cytometry. G2/M arrest was induced significantly with the highest concentration (4 mg $ml^{-1}$) of PS. RT-PCR was performed to study the correlation between G2/M arrest and transcription of cell cycle control genes. The anti-proliferative activity of CS and PS was accompanied by inhibition of cyclin B1, and Cdc 2 mRNA. Moreover, both CS and PS induced expression of the p53 tumor suppressor gene and the Cdk inhibitor p21. These results suggest that polysaccharides from S. herbacea have anti-cancer activity in human colon cancer cells.

• Journal of Ginseng Research
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• 제10권2호
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• pp.141-150
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• 1986

This study was attempted to screen the cytotoxic activity of petroleum ether ex- tract from panax ginseng root against human colon cancer cells. Two extracts of panax ginseng root, crude and partially purified, were used for this experiment. The crude extract was prepared by extraction with petroleum ether using Soxhlet aparatus for 12 to 15 hours from panax ginseng and the extract was partially purified by silicic acid column with mixture of petroleum ether: ethyl ether (70 : 30, v/v). Three species of human colon cancer cells, HRT-18, HCT-48 and HT-29, were maintained in DMEM (Dulbecco's modified Eagle medium), and the cells were cultured in DMEM containing serial concentration of the crude or partially purified fraction to observe the cytotoxic activity of the both extracts. The effects of incubation time and concentration of the both extracts in culture medium against the growth of the each cancer cell were determined. The results obtained are summarized as follows: 1. The doubling times of the HRT-18, HCT-48 and HT-29 cells were about 20, 24 and 22 hours, respectively. 2, The inhibitory effects of the crude extract on the growth of cancer cells were increased according to the rise of concentration of the extract and incubation time. 3. The inhibitory effect of partially purified fraction on the growth of HRT-18 cell was about 4 times stronger than that of the crude extract under same experimental condition. 4 The inhibitory effects of the crude and purified fraction on the growth of each cancer cell were shown difference by the kind of the cancer cell. In view of the above results, it could be said that the petroleum ether extract of panax ginseng root inhibited the division of the human colon cancer cell, in vitro.

• KSBB Journal
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• 제23권2호
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• pp.177-182
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• 2008

본 연구에서는 염생식물의 한 종류인 갯질경을 이용하여 생리활성물질에 의한 항발암성물질의 생성 및 생체방어물질로서의 유용성을 검토하기 위하여 갯질경의 추출물 및 용매별 분획물에서 AGS와 HT-29 인체암세포에 대한 증식억제효과를 검토하였다. 갯질경의 용매 추출물은 시료 처리 농도 증가에 따라 인체 위암 및 결장암세포의 생존율을 농도 의존적으로 감소시키는 경향을 보였다. 시료의 처리농도 $200\;{\mu}g/ml$에서 methanol과 methylene chloride 추출물은 인체 위암세포에 대하여 각각 66%와 71%의 암세포 증식억제율을, 인체 결장암세포에 대하여서는 각각 60%와 84%의 암세포 증식억제율을 보였다. 이러한 암세포 증식억제효과는 methanol 추출물보다는 methylene chloride 추출물에서 높게 나타났으며, AGS 인체 위암세포에서 보다도 HT-29 인체 결장암세포에서 다소 높은 효과를 나타내었다. 갯질경의 조추출물이 인체 암세포에 대하여 높은 증식억제효과를 나타내었으므로, 이들 추출물의 용매분획물을 이용하여 인체 암세포의 성장에 미치는 영향을 살펴 보았다. 그 결과 85%aq. MeOH 분획층에서 가장 높은 암세포 증식 억제효과를 보였으나, $H_2O$ 분획층에서는 50%이하의 낮은 억제율로 거의 효과를 보이지 않았다. AGS 인체 위암세포의 성장억제효과를 관찰한 결과, $200\;{\mu}g/ml$의 처리 농도에서 85% aq. MeOH, n-hexane 및 n-BuOH의 분획물이 약 80% 이상의 높은 증식 억제효과를 나타내었으며, HT-29 인체 결장암세포에 대하여서는 85%aq. MeOH과 n-hexane의 분획물에서 월등히 높은 증식억제효과가 관찰되었는데, 이 억제효과는 인체 위암세포에서 보다도 높은 저해효과를 보여주었다. 이상의 결과로부터 해양생물 중 염생식물의 한 종류인 갯질경의 높은 항암활성효과와 함께 식용 및 약용으로의 가치가 있는 새로운 생리활성물질의 소재군으로서의 가능성을 확인할 수 있었다.

• Journal of Nutrition and Health
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• 제40권2호
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• pp.147-153
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• 2007

In this study, we investigated the anticancer activity of the fin of Thunnus Thynnus (TT). TT was extracted with methanol (TTM), and then further fractionated into four subfractions by using solvent partition method, affording hexane (TTMH), methanol (TTMM), butanol (TTMB) and aquous (TTMA) soluble fractions. We determined the cytotoxicity of these four fractions in four kind of cancer cell lines, such as HepG2, MCF-7, B16-F10 and HT29 by MTT assay. The TTMM showed the strongest cytotoxic effect at the concentration of 150 ${\mu}g/mL$, displaying 95% on the HepG2 cell lines and 82% on MCF-7 cell line. The morphological changes such as membrane shirinking and blebbing of cells were also observed by TTMM treatment in HT29 cell. In addition, we observed that quinone reductase (QR) activity was elevated by only TTMM and TTMH treatments in HepG2 cell. QR activity was increased to around 2.0 and 1.8 times in TTMM and TTMH treated HepG2 cell at 100 ${\mu}g/mL$, respectively, compared to that in control. Although further studies are needed, the present work could suggest that the fin of TT has a potential to be usable as a chemopreventive agent against cancer.

• Journal of Microbiology and Biotechnology
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• 제17권7호
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• pp.1120-1126
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• 2007

This study was performed to screen probiotic bifidobacteria for their ability to bind and neutralize lipopolysaccharides (LPS) from Escherichia coli and to verify the relationship between LPS-binding ability, cell surface hydrophobicity (CSH), and inhibition of LPS-induced interleukin-8 (IL-8) secretion by HT-29 cells of the various bifidobacterial strains. Ninety bifidobacteria isolates from human feces were assessed for their ability to bind fluorescein isothiocyanate (FITC)-labeled LPS from E. coli. Isolates showing 30-60% binding were designated LPS-high binding (LPS-H) and those with less than 15% binding were designated LPS-low binding (LPS-L). The CSH, autoaggregation (AA), and inhibition of LPS-induced IL-8 release from HT-29 cells of the LPS-H and LPS-L groups were evaluated. Five bifidobacteria strains showed high levels of LPS binding, CSH, AA, and inhibition of IL-8 release. However, statistically significant correlations between LPS binding, CSH, AA, and reduction of IL-8 release were not found. Although we could isolate bifidobacteria with high LPS-binding ability, CSH, AA, and inhibition of IL-8 release, each characteristic should be considered as strain dependent. Bifidobacteria with high LPS binding and inhibition of IL-8 release may be good agents for preventing inflammation by neutralizing Gram-negative endotoxins and improving intestinal health.

• Asian Pacific Journal of Cancer Prevention
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• 제14권1호
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• pp.495-498
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• 2013

Epidermal growth factor receptor (EGFR) is over-expressed in several human cancers. This would suggest that inhibition of EGFR is a reasonable approach for cancer treatment. In this study we investigated EGFR blocking and its effects on the mediated signaling such as MAPK and STATb in HT29 cells. For this aim we used FITC-labeled EGFR antisense oligonucleotides encapsulated with PAMAM nanoparticles to inhibit EGFR expression. Cellular uptake of antisense was investigated by fluorescence microscopy and flow cytometry analysis. The effect of EGFR antisense on the expression of EGFR in HT29 cells was examined by real time PCR and Western blots, which showed that antisense encapsulated with PAMAM decreased the level of EGFR mRNA and protein. In addition, real time PCR results confirmed that EGFR inhibition had an effective role in the reduction of EGFR dependent downstream genes. In conclusion, EGFR antisense encapsulated with PAMAM nanoparticles down regulated EGFR and EGFR-mediated genes.