• Asian Pacific Journal of Cancer Prevention
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• 제15권8호
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• pp.3767-3772
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• 2014

Introduction: MicroRNAs have emerged as post-transcriptional regulators that are critically involved in tumorigenesis. This study was designed to explore the effect of miRNA 133b on the proliferation and expression of TBPL1 in colon cancer cells. Methods: Human colon cancer SW-620 cells and human colon adenocarcinoma HT-29 cells were cultured. MiRNA 133b mimcs, miRNA 133b inhibitors, siRNA for TBPL1 and scrambled control were synthesized and transfected into cells. MiR-133b levels in cells and CRC tumor tissue was measured by real-time PCR. TBPL1 mRNA was detected by RT-PCR. Cell proliferation was studied with MTT assay. Western blotting was applied to detect TBPL1 protein levels. Luciferase assays were conducted using a pGL3-promoter vector cloned with full length of 3'UTR of human TBPL1 or 3'UTR with mutant sequence of miR-133b target site in order to confirm if the putative binding site is responsible for the negative regulation of TBPL1 by miR-133b. Results: Real time PCR results showed that miRNA 133b was lower in CRC tissue than that in adjacent tissue. After miR-133b transfection, its level was elevated till 48h, accompanied by lower proliferation in both SW-620 and HT-29 cells. According to that listed in http://www.targetscan.org, the 3'-UTR of TBPL1 mRNA (NM_004865) contains one putative binding site of miR-133b. This site was confirmed to be responsible for the negative regulation by miR-133b with luciferase assay. Further, Western blotting and immunohistochemistry both indicated a higher TBPL1 protein expression level in CRC tissue. Finally, a siRNA for TBPL1 transfection obviously slowed down the cell proliferation in both SW-620 and HT-29 cells. Conclusion: MiR-133b might act as a tumor suppressor and negatively regulate TBPL1 in CRC.

• 생명과학회지
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• 제15권6호
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• pp.857-862
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• 2005

본 연구는 whole molecular-, 40 kDa-, 10 kDa poly-mannuronate를 대장암세포에 첨가하였을 때 분자량과 농도 에 따른 항암효과를 세포수준에서 검토하고자 하였다. 3종류의 인체 대장암세포인 HT-29, DLD-1, WiDr의 증식 억제를 살펴본 결과, HT-29 세포는 모든 polymannuronate $0.25\%$의 농도에서 $50\%$의 증식 억제 효과를 보였으며 저분자화 될 수 록 증식 억제 효과가 증가되었다. DLD-1 세포는 저분자화 정도와 농도증가에 따른 유의적인 차이를 보이지 않았고, WiDr 세포는 농도증가에 따른 유의적인 효과는 있었으나, 분자량에 의한 차이는 보이지 않았다. $0.25\%$의 농도에서 $50\%$의 증식 억제 효과를 보인 HT-29 세포의 DNA 합성능을 살펴본 결과, 증식 억제 효과와 유사한 결과를 보였으며, 형태학적인 변화 또한 동일한 것으로 나타났다. 이상의 결과로부터 polymannuronate는 HT-29 대장암세포의 증식억제에 유의적인 효과가 있는 것으로 나타났다. 따라서 polymannuronate는 대장암의 예방뿐 아니라 그 치료에도 도움이 되며 본 연구가 polymannuronate의 효과에 대한 기초적인 자료가 될 것으로 사료된다.

• 한국식품영양과학회지
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• 제28권1호
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• pp.240-245
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• 1999

Growth inhibitory effect of doenjang(Korean soypaste) methanol extracts in SRB assay using AGS human gastric adenocarcinoma cell, Hep 3B human hepatocellular carcinoma cell and HT 29 human colon cancer cell was studied. The treatment of doenjang methanol extracts(2mg/assay) to the AGS, Hep 3B and HT 29 cancer cells inhibited the growth of the cancer cells by 55%, 60%, and 71%, respectively. Doenjang methanol extracts exhibited the highest inhibitory effect among other soybean fermented foods and original materials in the SRB assay. In addition, to separate active compounds of doenjang methanol extracts, we fractionated the doenjang with hexane, methanol, dichloromethane, ethylacetate and butanol. Growth inhibitory effect on the AGS, Hep 3B, HT 29 and MG 63 cancer cells was the highest in the fractions of dichloromethane and ethylacetate among other solvent fractions of the doenjang. These results showed that some compounds contained in the fractions of dichloromethane and ethylacetate might play a role on the anticanceric effect of doenjang.

• 한국식품영양과학회지
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• 제35권9호
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• pp.1115-1120
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• 2006

육계 추출물의 대장암 세포에 미치는 영향을 HT-29 인체 결장암 세포주를 이용하여 조사하였다. 육계 추출물은 대장 정상세포인 CCD-112CoN의 성장에는 크게 영향을 미치지 않았으나, 대장암 세포인 HT-29의 성장은 농도, 시간에 의존적으로 크게 억제하였고, 대장암의 진행 과정에서 매우 중요한 COX-2 mRNA 발현량 및 $PGE_2$ 생성, cGMP 생성을 농도 의존적으로 억제하였다. 이상의 결과에서 육계 추출물은 정상세포에는 독성 없이 대장암 세포의 COX-2 및 $PGE_2$, cGMP 생성을 억제함으로써 결과적으로 대장암 세포의 성장을 감소시킨 것으로 판단된다. 따라서 육계는 부작용이 없는 대장암 예방 건강 보조식품으로서 개발 가능성이 있으며, 암세포 사멸에 대한 더욱 정확한 작용기전의 규명과 활성성분의 분리 및 정제, 응용법 등의 연구가 필요할 것으로 사료된다.

• Archives of Pharmacal Research
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• 제29권3호
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• pp.209-212
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• 2006

We investigated the effect of protein extract of Asterina pectinifera on the activity of 4 enzymes that may playa role in adenocarcinoma of the colon: quinone reductase (QR), glutathione Stransferase (GST), ornithine decarboxylase (ODC), and cyclooxygenase (COX)-2. QR and GST activity increased in HT-29 human colon adenocarcinoma cells increased that had been exposed to 4 concentrations of the protein extract (80, 160, 200, and $240{\mu}g/mL$). Additionally, 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced ODC activity decreased significantly in cells exposed to the extract in concentrations of $160{\mu}g/mL$ (p<0.05), $200{\mu}g/mL$ (p<0.005), and $240{\mu}g/mL$ (p<0.005). TPA-induced COX-2 activity also decreased in cells exposed to extract concentrations of 10, 20, 40, and $60{\mu}g/mL$. COX-2 expression was also inhibited in cells exposed to this extract. These results suggest that this protein extract of A pectinifera has chemopreventive activity in HT-29 human colon adenocarcinoma cells, and therefore, may have the potential to function as a chemopreventive agent in human colorectal cancer.

• Food Science and Biotechnology
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• 제16권2호
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• pp.260-264
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• 2007

Gleditsiae Semen (GS) has been used in both Korea and China as herbal medicine for the treatment of cephalalgia, catharsis, and other diseases. However, the apoptosis of GS against human cancer cells has not previously been investigated. The primary objective of this study was to determine the mechanisms inherent in GS-induced cytotoxicity and apoptosis, using methanolic extract of GS (GSE) in HT-29 human colon carcinoma cells. We found that GSE induced cytotoxicity in HT-29 cells in a dose-dependent manner, and this effect was verified via a lactate dehydrogenase release assay and a colony formation assay. In particular, HT-29 cells showed extensive cell death when treated with $50\;{\mu}g/mL$ of GSE; the calculated $IC_{50}$ value was $20\;{\mu}g/mL$. It induced characteristic apoptotic signs in HT-29 cells, including chromatin condensation and DNA fragmentation, occurring within 6-24 hr when the cells were treated at a concentration of $50\;{\mu}g/mL$. Interestingly, we detected the activation of caspase-3 and -9, but not caspase-8, and apoptotic bodies in GSE-treated HT-29 cells. Collectively, our results indicate that GSE induces apoptosis via a mitochondria-mediated apoptotic pathway, and these findings may be significant with regard to the development of a new drug for the treatment of human colon carcinoma cells.

• Preventive Nutrition and Food Science
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• 제7권3호
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• pp.250-254
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• 2002

The anticancer effects of leek (buchu in Korean) kimchi were evaluated in the human cancer cells: AGS gastric adenocarcinoma cells, HT-29 human colon adenocarcinoma cells and HL-60 leukemia cells. The leek kimchi (fermented for 6 days at 15$^{\circ}C$) was fractionated into 7 groups: methanol extract, hexane extract, methanol soluble extract MSE), dichloromethane (DCM) fraction (fr.), ethyl acetate fr., butanol fr. and aqueous fr. Most of the leek kimchi tractions inhibited the growth of AGS and HT-29 cancer cells in a dose dependent manner. In particular, the DCM fr. showed the highest inhibitory effect among the tractions. Treatment with the DCM fr. (0.1 mg/mL) reduced the survival rates of AGS and HT-29 cancer cells to 19% and 37% of the controls, respectively. Moreover the DCM fr. of the leek kimchi arrested G2/M phase in the cell cycle and induced apoptosis in HL-60 human promyelocytic leukemia cells. These results indicate that the leek kimchi exerted an anticancer effect on those human cancer cells, and that the DCM fr. arrested G2/M phase in the cell cycle and induced apoptosis in the leukemia cells.

• Nutrition Research and Practice
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• 제6권5호
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• pp.396-404
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• 2012

The aim of the study was to investigate the inhibitory effects of calcium against intestinal cancer in vitro and in vivo. We first investigated the effects of calcium treatment in HCT116 and HT29 human colon cancer cells. At the concentration range of 0.8-2.4 mM, calcium significantly inhibited cell growth (by 9-29%), attachment (by 12-26%), invasion (by 15-31%), and migration (by 19-61%). An immunofluorescence microscope analysis showed that the treatment with calcium (1.6 mM) for 24 h increased plasma membrane ${\beta}$-catenin but decreased nuclear ${\beta}$-catenin levels in HT29 cells. We then investigated the effect of dietary calcium on intestinal tumorigenesis in $Apc^{Min/+}$ mice. Mice received dietary treatment starting at 6 weeks of age for the consecutive 8 weeks. The basal control diet contained high-fat (20% mixed lipids by weight) and low-calcium (1.4 mg/g diet) to mimic the average Western diet, while the treatment diet contained an enriched level of calcium (5.2 mg calcium/g diet). The dietary calcium treatment decreased the total number of small intestinal tumors (by 31.4%; P < 0.05). The largest decrease was in tumors which were ${\geq}$ 2 mm in diameter, showing a 75.6% inhibition in the small intestinal tumor multiplicity (P < 0.001). Immunohistochemical analysis showed significantly reduced nuclear staining of ${\beta}$-catenin (expressed as nuclear positivity), but increased plasma membrane staining of ${\beta}$-catenin, in the adenomas from the calcium-treated groups in comparison to those from the control group (P < 0.001). These results demonstrate intestinal cancer inhibitory effects of calcium both in human colon cancer cells and $Apc^{Min/+}$ mice. The decreased ${\beta}$-catenin nuclear localization caused by the calcium treatment may contribute to the inhibitory action.

• BMB Reports
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• 제43권11호
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• pp.750-755
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• 2010

Crude Orostachys japonicus polysaccharide extract (OJP) was prepared by hot steam extraction. Polysaccharides (OJPI) were separated from OJP by gel filtration chromatography and phenol-sulfuric acid assay. The average molecular weight of the OJPI was 30-50 kDa. The anti-proliferative effect of OJPI on HT-29 human colon cancer cells was investigated via morphology study, cell viability assay, apoptosis assay, cell cycle analysis, and cDNA microarray. OJPI inhibited proliferation and growth of HT29 cells and also stimulated apoptosis in a dose- and time-dependent manner. In cell cycle analysis, treatment with OJPI resulted in a marked increase of cells in the G0 (sub G1) and G2/M phases. To screen for genes involved in the induction of cell cycle arrest and apoptosis, the gene expression profiles of HT-29 cells treated with OJPI were examined by cDNA microarray, revealing that a number of genes were up- or down-regulated by OJPI. Whereas several genes involved in anti-apoptosis, cell proliferation and growth, and cell cycle regulation were down-regulated, expression levels of several genes involved in apoptosis, tumor suppression, and other signal transduction events were up-regulated. These results suggest that OJPI inhibits the growth of HT-29 human colon cancer cells by various apoptosis-aiding activities as well as apoptosis itself. Therefore, OJPI deserve further development as an effective agent exhibiting anticancer activity.

• 생명과학회지
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• 제17권9호통권89호
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• pp.1232-1236
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• 2007

인체 암세포계(MG-63 골육암 세포, HT-29 인체 결장암세포, K-562 백혈암세포)를 이용하여 느릅나무 근피 메탄올 추출물, 열탕 추출물 및 즙액에 의한 암세포 성장에 미치는 효과를 검토하였다. 느릅나무 근피 메탄올 추출물, 열탕 추출물 및 즙액은 낮은 농도에서부터 인체 골육암 세포의 증식을 억제시켰다. 인체 결장암세포와 백혈암세포에서도 느릅나무 근피 메탄올 추출물, 열탕 추출물 및 즙액은 낮은 농도인 1 mg/ml에서부터 활성을 나타내어 40% 이상으로 암세포 증식 억제효과를 나타내어 앞서의 MG-63 골육암 세포에서 보다 그 증식 억제효과가 높은 것으로 나타났다. 느릅나무 근피 메탄올 추출물, 열탕 추출물 및 즙액을 골육암과 결장암세포에 투여한 2일 후에 세포내의 DNA 합성에 미치는 영향을 측정한 결과, 농도 의존적으로 암세포의 DNA 합성을 저해하는 것을 살펴 볼 수가 있었다. 따라서 느릅나무 근피 추출물은 인체 암세포 증식을 크게 억제하였으며 열탕 추출물에서도 항암활성 효과를 보여 느릅나무 근피 유래 생리활성 물질은 열에 안정한 것으로 여겨진다.