• Asian Pacific Journal of Cancer Prevention
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• 제15권23호
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• pp.10483-10487
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• 2015

Heptaphylline derivatives are carbazoles in Clausena harmandiana, a medicinal plant that is utilized for headache, stomach ache, and other treatments of illness. The present study examined the effects of heptaphylline and 7-methoxyheptaphylline on apoptosis of human colon adenocarcinoma cells (HT-29 cell line). Quantification of cell viability was performed using cell proliferation assay (MTT assay) and of protein expression through immunoblotting. The results showed that only heptaphylline, but not 7-methoxyheptaphylline, significantly significantly activated cleaved of caspase-3 and poly (ADP-ribose) polymerase (PARP-1) which resulted in HT-29 cell death. We found that heptaphylline activated BH3 interacting-domain death agonist (Bid) and Bak, proapoptotic proteins. In contrast, it suppressed X-linked inhibitor-of-apoptosis protein (XIAP), Bcl-xL and survivin, inhibitors of apoptosis. In addition, heptaphylline inhibited activation of NF-${\kappa}B$/p65 (rel), a regulator of apoptotic regulating proteins by suppressing the activation of Akt and $IKK{\alpha}$, upstream regulators of p65. The findings suggested that heptaphylline induces apoptosis in human colon adenocarcinoma cells.

• Asian Pacific Journal of Cancer Prevention
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• 제17권3호
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• pp.1103-1110
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• 2016

In the present study, we investigated the effects of motesanib (AMG 706), a multikinase inhibitor alone and in combination with DuP-697, an irreversible selective inhibitor of COX-2, on cell proliferation, angiogenesis, and apoptosis induction in a human colorectal cancer cell line (HT29). Real time cell analysis (RTCA, Xcelligence system) was used to determine the effects on colorectal cancer cell proliferation. Apoptosis was assessed with annexin V staining and angiogenesis was determined with chorioallantoic membrane model. We found that motesanib alone exerted antiproliferative, antiangiogenic and apoptotic effects on HT29 colorectal cancer cells. Combination with DUP-697 increased the antiproliferative, antiangiogenic and apoptotic effects. Results of this study indicate that motesanib may be a good choice in treatment of colorectal tumors. In addition, the increased effects of combination of motesanib with DuP-697 raise the possibility of using lower doses of these drugs and therefore avoid/minimize the dose-dependent side effects generally observed.

• 대한한의학방제학회지
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• 제32권2호
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• pp.111-120
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• 2024

Objectives : Intestinal epithelial cell damage is closely associated with various intestinal diseases, such as Inflammatory Bowel Disease (IBD), Celiac Disease and Gastroenteritis, and it plays a crucial role in the development and progression of intestinal diseases. Therefore, it is important to develop drugs that target protection of intestinal epithelial cells. Here, we aimed to investigated whether Bambusae Caulis in Liquamen (BCL) against t-BHP induced oxidative stress injury in human intestinal epithelial cells and to explore the underlying molecular mechanism. Methods : In this study, we performed MTT assay, measurement of ROS generation, and immunoblot analysis to determine the cytoprotective efficacy in HT29 cells (human colorectal adenocarinoma cell line with epithelial morphogy). Results : First, we checked that BCL was not cytotoxic up to concentration 30 ㎍/mL in HT29 cells. Then, we confirmed that BCL inhibited t-BHP-induced ROS and cell death. BCL also reversed the expression of proteins associated apoptosis. Next, to confirm the relationship between efficacy of BCL and Nrf2, we conducted experiments using siNrf2. Asresult, the effects of inhibiting ROS production and cell death of BCL was reversed by siNrf2. Conclusion : BCL prevents t-BHP-induced oxidative stress and apoptosis. And the efficacy of BCL is related to Nrf2 activation.

• IMMUNE NETWORK
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• 제13권5호
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• pp.213-217
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• 2013

Enterotoxigenic Bacteroides fragilis (ETBF) is a human gut commensal bacteria that causes inflammatory diarrhea and colitis. ETBF also promotes colorectal tumorigenesis in the Min mouse model. The key virulence factor is a secreted metalloprotease called B. fragilis toxin (BFT). BFT induces E-cadherin cleavage, cell rounding, activation of the ${\beta}$-catenin pathway and secretion of IL-8 in colonic epithelial cells. However, the precise mechanism by which these processes occur and how these processes are interrelated is still unclear. E-cadherin form homophilic interactions which tethers adjacent cells. Loss of E-cadherin results in detachment of adjacent cells. Prior studies have suggested that BFT induces IL-8 expression by inducing E-cadherin cleavage; cells that do not express E-cadherin do not secrete IL-8 in response to BFT. In the current study, we found that HT29/C1cells treated with dilute trypsin solution induced E-cadherin degradation and IL-8 secretion, consistent with the hypothesis that E-cadherin cleavage causes IL-8 secretion. However, physical damage to the cell monolayer did not induce IL-8 secretion. We also show that EDTA-mediated disruption of E-cadherin interactions without E-cadherin degradation was sufficient to induce IL-8 secretion. Finally, we determined that HT29/C1 cells treated with LiCl (${\beta}$-catenin activator) induced IL-8 secretion in a dose-dependent and time-dependent manner. Taken together, our results suggest that BFT induced IL-8 secretion may occur by the following process: E-cadherin cleavage, disruption of cellular interactions, activation of the ${\beta}$-catenin pathway and IL-8 expression. However, we further propose that E-cadherin cleavage per se may not be required for BFT induced IL-8 secretion.

• Asian Pacific Journal of Cancer Prevention
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• 제13권12호
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• pp.6349-6355
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• 2012

Previous studies showed that Math1 homologous to human Hath1 can cause mouse goblet cells to differentiate. In this context it is important that the majority of colon cancers have few goblet cells. In the present study, the potential role of Hath1 in colon carcinogenesis was investigated. Sections of paraffin-embedded tissues were used to investigate the goblet cell population of normal colon mucosa, mucosa adjacent colon cancer and colon cancer samples from 48 patients. Hath1 and Muc2 expression in these samples were tested by immunohistochemistry, quantitative real-time reverse transcription -PCR and Western blotting. After the recombinant plasmid, pcDNA3.1(+)-Hath1 had been transfected into HT29 colon cancer cells, three clones were selected randomly to test the levels of Hath1 mRNA, Muc2 mRNA, Hath1, Muc2, cyclin D1 and p27 by quantitative real-time reverse transcription-PCR and Western blotting. Moreover, the proliferative ability of HT29 cells introduced with Hath1 was assessed by means of colony formation assay and xenografting. Expression of Hath1, Muc2, cyclin D1 and p27 in the xenograft tumors was also detected by Western blotting. No goblet cells were to be found in colon cancer and levels of Hath1 mRNA and Hath1, Muc2 mRNA and Muc2 were significantly down-regulated. Hath1 could decrease cyclin D1, increase p27 and Muc2 in HT29 cells and inhibit their proliferation. Hath1 may be an anti-oncogene in colon carcinogenesis.

• Journal of Nutrition and Health
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• 제55권1호
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• pp.47-58
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• 2022

Purpose: Obesity and a high-fat diet (HFD) are risk factors for colorectal cancer. We have previously shown that luteolin (LUT) supplementation in HFD-fed mice markedly inhibits tumor development in chemically induced colon carcinogenesis. In this study, we evaluated the anticancer effect of LUT in the inhibition of cell proliferation in HFD-fed obese mice and HT-29 human colorectal adenocarcinoma cells grown in an adipocyte-derived medium. Methods: C57BL/6 mice were fed a normal diet (ND, 11.69% fat out of total calories consumed, n = 10), HFD (40% fat out of total calories consumed, n = 10), HFD with 0.0025% LUT (n = 10), and HFD with 0.005% LUT (n = 10) and were subjected to azoxymethane-dextran sulfate sodium chemical colon carcinogenesis. All mice were fed the experimental diet for 11 weeks. 3T3-L1 preadipocytes and HT-29 cells were treated with various doses of LUT in an adipocyte-conditioned medium (Ad-CM). Results: The weekly body weight changes in the LUT groups were similar to those in the HFD group; however, the survival rates of the LUT group were higher than those of the HFD group. Impaired crypt integrity of the colonic mucosa in the HFD group was observed to be restored in the LUT group. The colonic expression of proliferating cell nuclear antigen and insulin-like growth factor 1 (IGF-1) receptors were suppressed by the LUT supplementation in the HFD-fed mice. The LUT treatment (10, 20, and 40 µM) inhibited the proliferation and migration of HT-29 cells cultured in Ad-CM in a dose-dependent manner, as well as the differentiation of 3T3-L1 preadipocytes. Conclusion: These results suggest that the anticancer effect of LUT is probably due to the inhibition of IGF-1 signaling and adipogenesis-related cell proliferation in colon cancer cells.

• 한국미생물·생명공학회지
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• 제42권1호
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• pp.73-81
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• 2014

본 연구에서는 명아주과 수송나물속 솔장다리(Salsola collina Pall.) 추출물의 항산화 및 항암 활성을 분석하였다. 먼저 솔장다리의 에탄올 추출물의 DPPH radical scavenging activity를 분석한 결과, $IC_{50}$가 $4.82{\mu}g/ml$로 나타나 강한 항산화능을 보유하였음을 확인하였다. 또한 대장암 세포주(HT29), 폐암 세포주(A549), 간암 세포주(HepG2)를 사용하여 솔장다리 추출물의 암세포 사멸효과를 분석한 결과, $IC_{50}$가 각각 43.8, 64.1, $92.5{\mu}g/ml$로 강력한 세포사멸효과를 나타냈으며 특히 HT29에 대한 강한 사멸효과를 보였다. 솔장다리 추출물의 항암 활성 기전 분석을 위해 세포주기를 분석한 결과, 대장암세포인 HT29의 G2/M arrest를 유도하였으며 최고 농도인 $60{\mu}g/ml$까지 S기 세포수가 증가하였다. 세포주기관련 단백질의 발현 분석 결과, 솔장다리 추출물을 처리한 경우, G2기에서 M기로의 전이에 필수적인 단백질인 Cdc25C와 cyclin A의 발현이 감소되었고, 반면 Cdc25C와 Cdc2의 불활성화 형태인 p-Cdc25C, p-Cdc2는 증가하였다. 또한 p21과 Wee1의 발현도 증가되었다. 하지만 p53의 발현량은 변화가 없었다. 이러한 결과는 솔장다리 추출물을 처리한 경우, p53 비의존적으로 p21의 발현이 증가되어 cyclin A/Cdc2 complex의 활성이 조절되고, 이어서 G2/M phase의 check point에 작용하는 Wee1의 발현증가 및 Cdc25C, Cdc2의 인산화에 의한 불활성화를 통하여 G2/M arrest가 유도되는 것을 시사한다. 또한 솔장다리 추출물 처리에 의해 S기 진행을 조절하는 Cdk2의 발현량도 감소하여, cyclin A/Cdk2 complex가 감소되어 S기의 세포수가 증가한 것으로 보인다. 따라서 본 연구 결과를 통해 솔장다리 추출물이 높은 항산화 활성을 지니며 암세포의 세포주기를 조절하여 높은 항암 활성을 보유함을 확인하였다.

• KSBB Journal
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• 제27권4호
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• pp.227-231
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• 2012

Whole plants of Zostera asiatica were extracted twice with acetone/methylene chloride (A+M) and methanol (MeOH) in turn. The combined crude extracts were evaporated in vacuo and then the residue was partitioned between water and methylene chloride. The aqueous layer was fractionated into $H_2O$ and n-butanol and then the organic layer was also fractionated into 85% aq. MeOH and n-hexane, successively. The crude extracts and their solvent fractions were evaluated for their inhibitory effect on growth of human cancer cells AGS, HT-29, MCF-7, and HT-1080 cells by MTT reduction assay. Among samples tested, 85% aq. MeOH and n-hexane fractions showed strong cytotoxic effect against AGS, HT-29, and MCF-7 cells. On the other hand, for HT-1080 cell, 85% aq. MeOH fraction exhibited the strongest cytotoxic effect.

• 생명과학회지
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• 제20권9호
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• pp.1365-1372
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• 2010

본 연구에서는 마를 건조 후 분말화하여 실험에 사용하였으며, 건조 마 분말에 대한 생리활성에 대한 연구로 건조된 마를 유기용매로 추출하여 마 추출물과 분획물들의 $H_2O_2$에 의해 유도된 산화적 스트레스 및 인체 암세포(HT1080 인체 섬유육종세포, HT-29 인체 결장암세포)에 대한 증식 억제 효과에 대해 검토하고자 하였다. 마는 항비만, 항변비, 항돌연변이, 혈당 및 혈중 콜레스테롤 감소 등의 활성이 있어 건강식품으로 알려져 있다. $H_2O_2$에 의해 유도된 산화적 스트레스 저해효과를 알아보기 위하여 DCFH-DA assay를 행하였다. 건조 마 A+M 추출물을 농도별로 인체 섬유육종세포(HT1080)에 처리하였을 때 농도 의존적으로 세포 내 활성산소종을 크게 억제시켰다. 각 분획물들 중 85% aq. MeOH 분획물은 다른 분획물과 비교하여 높은 지질 과산화물 생성 억제 효과를 보였다. 인체 암세포에 대한 증식 억제 실험에서 A+M 및 MeOH 추출물은 농도 의존적으로 인체 섬유육종 및 결장암세포의 증식을 억제하였다. 마 추출물로부터 얻어진 n-hexane, 85% aq. MeOH 및 n-butanol (n-BuOH) 분획물들은 첨가농도 0.5 mg/ml 이상 처리했을 때 이들 두 암세포의 증식을 유의적으로 억제하였다(p<0.05). 본 연구 결과로 부터 건조 마의 암세포 증식 억제 효과는 마의 A+M 추출물이 MeOH 추출물에 비해 높았고, 분획물들의 경우에는 water 분획물을 제외한 분획물들의 암세포 억제 효과가 높았다. 특히 85% aq. MeOH 분획물은 인체 암세포 증식 억제 효과뿐만 아니라 세포 내 활성산소종 감소시키는 효과가 높아 이들 분획물에 활성물질이 있을 것으로 추정된다.

• 한국식품영양과학회지
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• 제37권7호
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• pp.826-833
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• 2008

김 가공 후 발생되는 자투리 김으로부터 porphyran을 추출(13.9% 수율)하고, 이를 청국장에 첨가하여 porphyran-청국장을 제조하고 이의 항암활성을 조사하였다. Crude porphyran의 암세포에 대한 증식억제효과는 AGS와 HT-29에서 모두 농도가 증가할수록 농도 의존적으로 높은 억제율을 나타내었다. Crude porphyran의 정상세포 BJ에 대한 세포 독성은 거의 없는 것으로 관찰되었으며 0.25 mg/mL의 농도까지는 농도 의존적으로 세포성장효과를 보였다. Bacillus subtilis DJI으로 제조한 청국장의 물 추출물과 메탄올 추출물의 세포증식에 미치는 효과를 살펴보았다. 정상세포 BJ에 대해 청국장 물 추출물의 경우 0.5 mg/mL의 농도에서는 최고 153%의 생존율을 나타냈으며, 메탄올 추출물의 경우 동일 농도에서 140%의 생존율을 보여 청국장이 정상세포에 대해 세포성장효과가 있는 것으로 관찰되었다. Porphyran-청국장의 정상세포 BJ에 대한 세포독성을 관찰한 결과 물, 메탄올 추출물 모두 독성이 없었으며, 물 추출물의 경우 1.0 mg/mL 농도 처리 시 160%, 메탄올 추출물의 경우 0.25 mg/mL에서 최대 127%의 생존율을 보이므로 porphyran-청국장이 정상세포에 대해 세포독성이 없는 것으로 관찰되었다. 청국장 추출물과 porphyran-청국장 추출물의 암세포 증식 억제효과를 농도 1.0 mg/mL 구간에서 비교해보았을때 porphyran을 첨가하지 않은 청국장 물 추출물에서 최고 17%, 메탄올 추출물은 24%의 AGS 세포증식억제효과를 나타내는 것에 비해, porphyran-청국장의 물 추출물과 메탄올 추출물은 최고 23%와 38%의 세포증식 억제효과를 각각 나타내었다. 장암세포 HT-29에 대해서는 porphyran을 첨가하지 않은 청국장 물 추출물에서 19%, 메탄올 추출물은 27%의 세포증식억제효과를 나타내는 것에 비해 porphyran-청국장의 물 추출물과 메탄올 추출물은 최고 27%, 32%를 각각 나타내었다. 이로부터 청국장과 porphyran-청국장의 물 추출물보다 메탄올 추출물이 더 높은 항암활성을 나타내며, 청국장보다 porphyran-청국장이 $1.2{\sim}1.5$배 더 높은 암세포 성장 억제율을 나타냄을 알 수 있었다. 본 실험에서 김으로 부터 유용한 기능을 가지고 있는 porhyran을 추출하여 항암 활성 기능을 가지는 porphyran-청국장을 제조하여 그 우수성을 확인하였다. 앞으로 이러한 기능성 식품을 널리 이용하여 해조가공산업에 기여하고 부가가치를 향상시켜 새로운 소비를 창출할 수 있을 것으로 기대된다.