• 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.

• 생명과학회지
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• 제22권1호
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• pp.55-61
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• 2012

Akt는 세포의 증식과 분화에 관여하며 많은 암종에서 과발현되어 있다는 것이 보고되었다. 본 연구에서는 Akt의 조절을 통한 셀레늄의 HT-29 세포의 세포증식억제 시너지효과를 확인하였다. 셀레늄을 농도별과 시간별로 처리하였을 때 HT-29 세포의 증식이 억제되었고, apoptosis가 일어남을 확인하였다. 셀레늄을 농도별로 처리하여 Western blotting 및 immunofluorescence를 실시한 결과 Akt의 인산화가 저해되었고 COX-2의 발현도 저해되었다. 또한 Akt 저해제인 LY294002를 처리한 결과, HT-29 대장암세포의 증식이 억제되었으며, LY294002를 셀레늄과 병행처리하였을 때 셀레늄에 의한 세포증식억제 효과가 더 강하게 나타나는 것을 확인하였다. Akt siRNA에 의한 Akt의 불활성화는 non-transfected 세포에 비하여 HT-29 세포의 성장을 더 강하게 억제하였으며, Akt가 불활성화 되었을 때 COX-2의 발현 역시 non-transfected 세포에 비하여 감소된 것을 확인하였다. 따라서 HT-29 세포에서 셀레늄의 세포증식억제 효과는 Akt와 COX-2 신호분자의 조절을 통해 일어나며, Akt 의 저해는 셀레늄의 대장암세포증식 억제에 시너지 효과를 나타냄을 확인하였다.

• 한국식품영양과학회지
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• 제44권3호
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• pp.338-346
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• 2015

본 연구에서는 최근 식생활의 서구화로 인해 발병률이 급증하고 있는 대장암의 진행을 억제하거나 감소시키고 인체 대장암 세포인 HT-29의 증식을 억제하며, 세포사멸을 유도하는 천연소재를 알아보기 위해서 flavonoid가 HT-29 인체 대장암 세포의 apoptosis 유도 및 기전에 미치는 영향을 알아보았다. MTT assay 결과 apigenin, rutin, naringenin, myricetin을 $100{\mu}M$ 농도로 처리하였을 때 62.71, 75.78, 74.24, 77.61%로 이 중 naringenin이 대장암 세포 성장에 억제 효과가 가장 높은 실험 결과를 나타내었다. Caspase-3 activity에서는 naringenin이 241.46%로 가장 높은 활성을 나타내었다. 이를 바탕으로 세포사멸과 관련된 유전자를 확인하고자 대장암 세포에 flavonoid인 apigenin, rutin, naringenin, myricetin에 $100{\mu}M$ 농도로 처리한 후 RTPCR을 실시한 결과, 세포사멸의 주요한 조절인자인 Bcl-2 family 단백질 중 Bcl-2는 rutin에 의해 감소되었고 Bax는 myricetin에 의해 증가하였으며, p53은 naringenin이 높게 발현되었다. 또한 western blotting을 통해 flavonoid인 apigenin, rutin, naringenin, myricetin에 $100{\mu}M$ 농도로 처리한 결과, Bcl-2 family 단백질과 더불어 세포사멸 조절에 중요한 역할을 하는 활성형인 cleaved caspase-3은 모두 증가하였고, 그중 myricetin이, PARP은 naringenin, E-cadherin은 rutin이 각각 높은 발현 양상을 나타내었다. 이번 실험 결과를 통해 flavonoid가 세포사멸의 주요한 조절 인자인 Bcl-2 family 단백질의 발현이나 caspase의 활성 등을 조절하여 암세포 사멸인자인 Bcl-2의 발현은 감소시키고 Bax, p53, PARP의 발현을 증가시키는 것을 통해 대장암 세포의 apoptosis를 유도하였다. 또한 암세포의 전이와 관련된 E-cadherin의 발현도 조절하는 것을 관찰하였다. 이상의 연구를 통해 flavonoid가 대장암 세포의 증식을 억제하는 효과가 있음을 확인하였으며, 세포사멸과 관련된 기전을 규명하였다. 이를 기초자료로 일상에서 쉽게 섭취할 수 있는 식품에 많이 존재하며 비교적 독성과 부작용이 적은 flavonoid를 이용한 천연 항암제 개발 가능성을 제시하였고, 추후 대장암의 암예방제 및 암치료제로 개발될 수 있도록 추가 연구 수행이 필요할 것으로 사료된다.

• 대한한방내과학회지
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• 제31권2호
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• pp.273-289
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• 2010

Objectives : To investigate the anti-cancer effect of Baekduong-tang(BDOT) against cancer cells, the signaling pathway of apoptosis was explored in human colon cancer cells. Materials and Methods : Human colon cancer cell lines, including HT-29 and HCT-116 cells, were used. Cell viability was measured by MTT assay. Apoptosis was determined by DAPI nuclei staining and flow cytometry in HCT-116 cells treated with 0.25 mg/$m{\ell}$ Baekduong-tang for 48 hrs. Results : Baekduong-tang induced the apoptosis of p53 positive HCT-116 cells with G2/M phase arrest. Treatment with Baekduong-tang led to increased expression and phosphorylation of p53 and decreased expression of CDK2 and CDK6 in HCT-116 cells. It also activated caspase-3 through caspase-10 and caspase-9 activation. Finally, Baekduong-tang induced production $H_2O_2$, superoxide anion ($O_2^-$) and NO and modulated proteins expression including SOD, NOS, Bax and Bcl-2. Conclusions : These results indicate Baekduong-tang induces apoptotic death of HCT-116 cells through G2/M phase arrest and disturbance of intracellular redox status in a p53-dependent manner.

• 대한의생명과학회지
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• 제19권3호
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• pp.224-232
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• 2013

Cucurbitacins are a natural cell-permeable triterpenoid compound isolated from Cucurbitaceae and Cruciferae. Cucurbitacins have been used as folk medicine because of their anti-inflammatory and analgesic effects. In the present study, we investigate the anti-cancer effects of cucurbitacin I on colitis-associated colon carcinogenesis induced by azoxymethane (AOM)/dextran sodium sulfate (DSS) in BALB/c mice. Cucurbitacin I treatment attenuated loss of body weight and decreased the number of colon tumors. Western blot analysis showed that cucurbitacin I treatment significantly inhibited the protein expression of inducible nitric oxide synthase (iNOS), tumor necrosis factor (TNF)-${\alpha}$ and interleukin (IL)-6. These results suggest that cucurbitacin I suppressed inflammatory reaction and tumor development in colitis-associated colon carcinogenesis.

• 한국식품영양학회지
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• 제22권4호
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• pp.542-547
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• 2009

Brine mineral water(BMW) has recently gained attention as a new water resource due to its biological activities. In this study, BMW from the Geumjin area(Gangneung-city, Korea) was evaluated for its growth inhibition, anti-metastasis and anti-angiogenesis activity against cancer cells. The in vitro cytotoxicity was measured by CCK assay, and the anti-metastasis activity was estimated by lung metastasis in vivo. The in vitro incubation of mouse splenic cells with BMW that had been diluted more than 4-fold showed no effect on the cell growth when compared to a control group. Additionally, BMW inhibited the growth of the EL-4, L5178Y-R and colon26-M3.1 cancer cell lines in a dose-dependent manner. In vivo evaluation of the anti-metastasis activity of BMW in BALB/c mice inoculated with the colon26-M3.1 cell line revealed dose-dependent inhibition in response to treatment with samples that were diluted by up to 9 times. Finally, treatment with BMW effectively suppressed the growth of vascular endothelial growth factor(VEGF) added human umbilical vein endothelial cells. Overall, these results suggest that BMW has anti-cancer activity.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 추계학술대회
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• pp.101-101
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• 2018

Resveratrol is a polyphenolic compound, which is a naturally occurring phytochemical and is found in a variety of plants, including food such as grapes, berries and peanuts. Although several studies have demonstrated that resveratrol possesses anti-cancer activity against various types of human cancer, the molecular mechanisms of resveratrol-mediated overcome drug resistance potential are unclear. In this study, we determined whether resveratrol attenuates drug resistance responses in 5-fluorouracil-resistant colon cancer (SNUC5/5-FUR) cells. Treatment with resveratrol significantly enhanced apoptosis in a concentration-dependent manner, which was associated with the modulation of anti- and/or pro-apoptotic protein expression, activation of caspases and activation of AMP-activated protein kinase. Resveratrol treatment also increased the induction of autophagy through up-regulation of autophagy-related genes such as Microtubule-associated protein 1A/1B-light chain 3, P62 and beclin-1. However, blocking of autophagy by bafilomycin A1 reduced apoptotic cell death, suggesting that resveratrol-induced autophagy functions as a cell death mechanism in SNUC5/5-FU cells. Although the further studies are needed, these findings suggest that resveratrol may have therapeutic potential to overcome drug resistance in colon cancer patients.

• 생명과학회지
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• 제24권9호
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• pp.1012-1018
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• 2014

제주조릿대(Sasa quelpaertensis Nakai)는 한라산에 넓게 분포되어 자생하는 식물로 최근 연구에서 항염증, 항당뇨, 항산화, 항암 효능을 가지는 것으로 알려져 있으나 대장암에서의 항암 효능 및 그에 따른 mechanism에 대해서는 명확히 밝혀지지 않았다. 본 연구에서는 인간 대장암 HT-29 세포를 대상으로 제주조릿대에 의한 항암작용과 기전에 대해 조사하였다. 제주조릿대에 의한 HT-29 세포의 증식 억제가 apoptosis 유도와 연관성이 있음을 DNA fragmentation와 flow cytometry 분석에 의한 sub-G1기의 세포빈도의 증가로 확인하였다. 제주조릿대에 의한 apoptosis 유발은 HT-29 세포의 S arrest 현상을 동반하였을 뿐만 아니라 발생한 산화질소의 증가와 anti-apoptotic factor인 IAP family (survivin, XIAP, cIAP-1, cIAP-2) 발현이 감소함으로써 촉진되었음을 확인할 수 있었다. 이러한 결과들은 제주조릿대가 대장암에 대한 치료제로서의 사용 가능성을 확인할 수 있었지만 이를 입증하기 위해서는 더 자세한 항암기전에 관한 연구가 진행되어야 한다고 사료된다.

• 약학회지
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• 제48권3호
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• pp.189-196
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• 2004

Matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) playa key role in tumor invasion and metastasis. As an inhibitor of MMP-2, TIMP-2 is known to block both the invasive and metastatic behavior of cancer cells, and decrease tumor growth activity. We performed this study to investigate the effects of TIMP-2 over-expression induced by retroviral mediated gene transfer in vitro and in vivo. The human colon cancer cell line SW480 was transfected with the retroviral vector encoding TIMP-2. The effects of TIMP-2 over-expression were analyzed by invasion assay and gelatinase activity test in colon cancer cells and tumorigencity in nude mice. In evaluation of the transfection efficiency of the retroviral vector encoding TIMP-2 in colon cancer cells, we confirmed up-regulation of TIMP-2 expression dependent on the time of cell culture. In addition, inhibition of MMP-2 expression in SW480/TIMP-2 was shown by gelatin zymography. In the in vitro invasion assay SW480/TIMP-2 inhibited the invasiveness on matrigel coated with collagen. To determine whether TIMP-2 can modulate in vivo tumorigenicity and metastasis, SW480/TIMP-2 cells were injected subcutaneously in nude mice. The tumor mass formation of SW480/TIMP-2 cells in nude mice was markedly decreased compared to nontransfected cancer cells. These results showed that colon cancer cells transfected with the retroviral vector encoding TIMP-2 inhibits the invasiveness in vitro and tumorigenicity in vivo.

• 한국미생물·생명공학회지
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• 제41권3호
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• pp.379-382
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• 2013

Rhanella aquatilis AY2000 균주가 생산하는 일종의 당단백질인 항효모성 물질 (AYS)에 대한 항암활성을 조사하기 위해 in vitro에서 암세포에 대한 AYS의 세포독성을 조사하였다. 그 결과 AYS는 인체의 Jurkat T 세포와 마우스의 sarcoma 180 세포에 대해서는 세포독성을 나타내지 않았으나, 인체 대장암세포인 colon cancer TH20 세포에는 세포독성을 나타내었다. 또한 이 AYS는 62.5에서 500 ${\mu}g/ml$까지 농도의존적으로 인체대장암세포에 대해 세포독성을 증가시켰다. 뿐만 아니라 이 AYS와 시판 항암제를 혼합하여 처리한 결과 시판 항암제를 단독으로 처리한 것 보다 인체대장암세포에 대한 항암효과가 더욱 상승되었다.