• 대한의생명과학회지
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• 제17권1호
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• pp.85-88
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• 2011

Tumor angiogenesis, which is essential for tumor growth and tumor metastasis, depends on angiogenic factors produced by tumor cells and/or infiltrating cells such as endothelial cells and immune cells in tumor tissue. Previously, we reported that licochalcone A (LicA), an important bioactive compound of Glycyrrhiza inflate, suppresses angiogenesis, tumor growth and metastasis. In this study, we evaluated the effect of LicA on angiogenin production in colon cancer cells because angiogenin is an essential factor to regulate angiogenesis and tumor progression. When we examined the angiogenin levels in three human colon cancer cells, HT-29, SW480 and Caco-2, LicA treatment significantly reduced the amounts of angiogenin among three cancer cell lines. In an in vivo study in which mice were implanted with HT-29 cells, oral administration of LicA reduced angiogenin in tumor tissues when compared with vehicle-administered mice. These results suggest that reduced angiogenin in response to LicA treatment may play essential role to inhibit tumor growth, angiogenesis as well as metastasis.

• BMB Reports
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• 제42권5호
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• pp.277-280
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• 2009

We examined the effects of polysaccharides extracted from Asterina pectinifera on the activities of quinone reductase (QR), glutathione S-transferase (GST), ornithine decarboxylase (ODC), cyclooxygenase (COX)-2 and glutathione (GSH) levels in HT-29 human colon adenocarcinoma cells. We found that the polysaccharides extract induced QR activity in a dose-dependent manner over a concentration range of $20-60\;{\mu}g/ml$ and increased GST activity as much as 1.4-fold over controls. GSH levels were increased 1.3- and 1.5-fold with the extract at 40 and $60\;{\mu}g/ml$, respectively. The activity and protein expression of ODC in 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced colon cancer cells was inhibited by the extract. The polysaccharides suppressed TPA-induced prostaglandin (PG) production. These data indicate that polysaccharides from A. pectinifera increase phase II detoxification enzyme activity and inhibit ODC and COX-2 activities in HT-29 human colon adenocarcinoma cells. Consequently, this effect may contribute to the protective effect of polysaccharides from A. pectinifera against colon cancer.

• Food Science and Biotechnology
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• 제16권1호
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• pp.167-170
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• 2007

(-)-Epigallocatechin-3-gallate (EGCG), a mai or tea catechin has been shown to have many interesting biological activities. In the present study, we studied the effects of green tea catechins, EGCG metabolites, and black tea theaflavins on accumulation of EGCG in HT-29 human colon cells. Intracellular levels of [$^3H$]-EGCG were not changed significantly in the presence of other tea catechins including (-)-epicatechin, (-)-epigallocatechin, and (-)-epicatechin-3-gallate. EGCG methyl metabolites and EGCG 4"-glucuronide did not affect cellular levels of [$^3H$]-EGCG. Black tea theaflavins and theasinensin A (TsA), an EGCG oxidative dimer, however, significantly decreased cellular accumulation of EGCG in HT-29 cells by 31-56%. This decrease was more pronounced when cells were incubated in the presence of theaflavin-3',3"-digallate (TFdiG) or TsA. When EGCG was added separately from TFdiG or TsA, the accumulation of EGCG in HT-29 cells was also significantly decreased regardless of when TFdiG or TsA was added during the uptake study (p<0.01). The results suggest that theaflavins and TsA may interrupt EGCG absorption through the gastrointestinal epithelium.

• Food Science and Biotechnology
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• 제17권6호
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• pp.1265-1271
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• 2008

Adlay bran is a waste product previously thought to have no commercial value, Its methanolic extract was fractionated using n-hexane (ABM-Hex), ethyl acetate (ABM-EtOAc), 1-butanol (ABM-BuOH), and water (ABM-$H_2O$). The ABM-EtOAc fraction exhibited a strongest inhibition against growth of human lung cancer cell A549 and human colorectal carcinoma cells HT-29 and COLO 205. Inhibition of cell cycle progression at $G_0/G_1$ transition, increase of cells at the sub-$G_1$ phase, and DNA ladders were observed in cells treated with ABM-EtOAc. The ABM-BuOH fraction showed the strongest inhibition of proinflammatory cytokines tumor necrosis factor (TNF)-$\alpha$ and interlukin (IL)-$1{\beta}$ in stimulated RAW 264.7 macrophages. Further, ABM-EtOAc and ABM-BuOH inhibited cyclooxygenase (COX)-2 expression in A549 and HT-29 carcinoma cells, while COX-l expression was not affected. These results reveal that both ABM-EtOAc and ABM-BuOH may aid the prevention of cancers and the applications in cancer chemotherapy.

• 한국식생활문화학회지
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• 제24권6호
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• pp.749-755
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• 2009

We investigated the chemical components of red onion powder dried using the low temperature vacuum method and the inhibitory effects of solvent extracts of the dried red onion powder on the growth of HT-1080 human fibrosarcoma and HT-29 human colon cancer cells and $H_2O_2$-induced oxidative stress. The moisture content of the dried red onion powder was 17.95%, while the vitamin C content was 96 mg/100 g and the total phenols content was 39.1 mg/mL. The inhibitory effects of acetone with methylene chloride (A+M) and methanol (MeOH) extracts of the red onion powder on the growth of HT-1080 and HT-29 cancer cells increased in a dose dependent manner (p<0.05). The inhibitory effect was greater on the growth of HT-29 cells, while the A+M extracts had a higher inhibitory effect than the MeOH extracts. Treatment with the hexane, 85% aq. methanol, butanol and water fractions of the extract led to significant inhibition of the growth of both cancer cell lines (p<0.05). Among the fractions, the hexane and 85% aq. methanol fractions showed a greater inhibitory effect. To determine the protective effect on $H_2O_2$-induced oxidative stress, a DCFH-DA (dichlorodihydrofluorescin diacetate) assay was conducted. All fractions, including the crude extracts of dried red onion, appeared to lead to a significant reduction in the levels of intracellular reactive oxygen species (ROS), and these reductions occurred in a dose dependent fashion (p<0.05). Among the fractions, the 85% methanol fraction showed the greatest protective effect on the production of lipid peroxides.

• Applied Biological Chemistry
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• 제49권3호
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• pp.248-253
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• 2006

조각인(Gleditsiae Semen)은 동아시아 지역에서 장 기능을 향상시키는 등의 여러 가지 민간요법으로 사용되는 한약재이다. 본 연구에서는 조각인의 항암활성을 검색하기 위한 목적으로 methanol, ethanol, acetone으로 추출한 뒤, 인간 유래의 대장암 세포주 HT-29에 처리하여 각 추출물의 암세포 성장 억제능을 확인하기 위하여 MTT-dye reduction assay를 이용하여 실험하였다. 그 결과, methanol 추출물에서 가장 높은 세포독성을 확인할 수 있었고, ethanol 추출물에서도 비교적 높은 암세포 성장 억제능을 볼 수 있었으나, acetone 추출물에서는 거의 세포독성이 나타나지 않았다. 그리고 추출물들을 $25\;{\mu}g/ml$의 농도로 암세포에 처리하여 위상차 현미경으로 세포주의 형태학적 변화를 관찰한 결과에서도 methanol 추출물에서의 활성이 가장 강력하게 나타남을 알 수 있었다. 조각인의 항암활성물질들의 용매특성을 알아보기 위하여 methanol 추출물을 극성도에 따라 용매분획 하여 분리한 각 분획물의 세포독성을 확인한 결과, water층 분획물에서 농도 의존적으로 세포독성을 나타냄을 확인하였다. 또한 조각인의 water층 분획물은 또 다른 대장암 세포주인 SW620에 대해서도 대조군에 비해 $25\;{\mu}g/ml$의 농도에서 2배 정도 높은 항암활성을 나타내고 있음을 MTT reduction assay와 LDH release assay를 통하여 확인 할 수 있었다.

• 한국미생물·생명공학회지
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• 제41권4호
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• pp.425-432
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• 2013

본 연구에서는 인체 대장암세포인 HT29를 사용하여 포황 메탄올 추출물(Methanol extract of Typha orientalis, METO)의 항암 활성 및 그 분자적 기전에 관하여 분석하였다. 먼저 METO가 다양한 암세포의 증식에 미치는 영향을 분석한 결과, 인체 대장암 세포, 폐암 세포, 간암 세포 등의 세포증식을 억제하였으며 그 중에서도 대장암 세포인 HT29에 대해 강한 세포 증식 억제 효과를 나타내었다. METO에 의한 세포 증식 억제 기전을 분석하기 위하여 Flow cytometry analysis를 수행한 결과, METO 농도의존적으로 HT29 세포의 G2/M기 세포분포가 증가하고 아울러 apoptosis 유발군인 SubG1기 세포분포가 증가하는 것을 확인할 수 있었다. METO에 의한 HT29 세포의 G2/M arrest는 Cdc2의 inactive form인 phospho-Cdc2의 증가에 의한 G2/M checkpoint 관련 단백질의 활성억제에 의한 것이라 사료된다. 이러한 phospho-Cdc2의 증가는 METO에 의해 발현이 증가된 Wee1 kinase와 발현이 감소된 Cdc25C phosphatase에 의해 야기된 것임을 확인하였다. 또한 METO에 의한 HT29의 apoptosis 유도에 관한 분자적 기전 분석을 위해 Western blot analysis를 수행한 결과, METO 농도가 증가할수록 종양 억제 유전자인 p53, death receptor인 FAS, Bcl-2 family 중 pro-apoptotic 단백질인 Bax 및 cytosolic cytochrome C의 발현이 증가되고, Caspase-3가 활성화되어 단편화된 Caspase-3의 증가가 관찰되었다. 또한 활성화된 Caspase-3의 기질 단백질인 PARP의 단편화가 일어나 apoptosis가 유도되는 것을 알 수 있었다. 이상의 결과들로부터 METO는 인체 대장암세포 HT29의 G2/M arrest 및 apoptosis 유도에 의한 항암활성을 보유함을 확인하였다.

• Asian Pacific Journal of Cancer Prevention
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• 제17권5호
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• pp.2637-2641
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• 2016

Tumor necrosis factor ($TNF-{\alpha}$), an inflammatory cytokine that plays an important role in the control of cell proliferation, differentiation, and apoptosis, has previously been used in anti-cancer therapy. However, the therapeutic applications of $TNF-{\alpha}$ are largely limited due to its general toxicity and anti-apoptotic influence. To overcome this problem, the present study focused on the effect of active constituents isolated from a medicinal plant on $TNF-{\alpha}$-induced apoptosis in human colon adenocarcinoma (HT-29) cells. Nimbolide from Azadirachta indica was evaluated for cytotoxicity by methyl tetrazolium 3-[4,5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay and phase contrast microscopy. Effects on apoptotic signaling proteins were investigated using Western blot analysis. Nimbolide showed cytotoxicity against HT-29 cells that was significantly different from the control group (p<0.01), a concentration of $10{\mu}M$ significantly inducing cell death (p<0.01). In combination with $TNF-{\alpha}$, nimbolide significantly enhanced-induced cell death. In apoptotic pathway, nimbolide activated c-Jun N-terminal kinase (JNK) phosphorylation, BH3 interacting-domain death agonist (Bid) and up-regulated the death receptor 5 (DR5) level. In the combination group, nimbolide markedly sensitized $TNF-{\alpha}$-induced JNK, Bid, caspase-3 activation and the up-regulation of DR5. Our findings overall indicate that nimbolide may enhance $TNF-{\alpha}$-mediated cellular proliferation inhibition through increasing cell apoptosis of HT-29 cells by up-reglation of DR5 expression via the JNK pathway.

• Asian Pacific Journal of Cancer Prevention
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• 제15권7호
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• pp.3113-3121
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• 2014

Colorectal cancer remains one of the most common types of cancer and a leading cause of cancer death worldwide. In this study, we aimed to investigate effects of DuP-697, an irreversible selective inhibitor of COX-2 on colorectal cancer cells alone and in combination with a promising new multi-targeted kinase inhibitor E7080. The HT29 colorectal cancer cell line was used. Real time cell analysis (xCELLigence system) was conducted to determine effects on colorectal cell proliferation, angiogenesis was assessed with a chorioallantoic membrane model and apoptosis was determined with annexin V staining. We found that DuP-697 alone exerted antiproliferative, antiangiogenic and apoptotic effects on HT29 colorectal cancer cells. For the antiproliferative effect the half maximum inhibition concentration ($IC_{50}$) was $4.28{\times}10^{-8}mol/L$. Antiangiogenic scores were 1.2, 0.8 and 0.5 for 100, 10 and 1 nmol/L DuP-697 concentrations, respectively. We detected apoptosis in 52% of HT29 colorectal cancer cells after administration of 100 nmol/L DuP-697. Also in combination with the thyrosine kinase inhibitor E7080 strong antiproliferative, antiangiogenic and apoptotic effects on HT29 colorectal cancer cells were observed. This study indicates that DuP-697 may be a promising agent in the treatment of colorectal cancer. Additionally the increased effects observed in the combination with thyrosine kinase inhibitor give the possibility to use lower doses of DuP-697 and E7080 which can avoid and/or minimize side effects.

• 생명과학회지
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• 제24권3호
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• pp.234-241
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• 2014

용규(Solanum nigrum)에 함유된 여러 성분 중 항암 활성이 있는 물질을 분리 및 규명하고자 용규를 MeOH로 추출하고 이 추출물을 $CH_2Cl_2$, EtOAc, n-BuOH, $H_2O$로 유기용매 계통분획 하였다. 이 분획물 중 $H_2O$ 획분이 가장 뛰어난 세포 독성 효과를 보였으며 이를 다시 용매별($H_2O$, 40% MeOH, 60% MeOH, MeOH)로 Diaion HP-20 column chromatography하여 4개의 fraction으로 나누었다. 이 fraction 중 $H_2O$ fraction에서 가장 뛰어난 세포 독성효과를 보였으며 몇 차례의 column chromatography를 행하여 얻어진 활성 획분을 GC-MS 및 FAB-MS 분석하였다. 분자량이 416이었으며 GC-Mass를 통하여 library search를 행한 결과 분자식 $C_{28}H_{48}O_2$인 Des-N-26-methylenedihydrotomatidine으로 확인되었다. Des-N-26-methylene-dihydrotomatidine과 비슷한 구조를 가지고 있는 tomatine과 tomatidine의 HT29 세포에 대한 세포독성효과를 확인한 결과, tomatine보다는 활성이 낮았으나 tomatidine 보다는 40 ${\mu}g/ml$ 이상의 농도에서 높은 세포독성 효과를 나타내었다. Des-N-26-methylene-dihydrotomatidine은 지금까지 학계에 보고되어 있지 않은 물질로, 본 연구에서 최초로 용규에서 Des-N-26-methylene-dihydrotomatidine을 분리하였고, 암세포에 대한 세포 독성 효과를 보고하였다. Des-N-26-methylene-dihydrotomatidine의 정상세포에서의 세포 독성 및 작용 기전 등의 추가 연구를 통해 천연 생리 활성 물질 소재로 활용 가능할 것으로 사료된다.