• BMB Reports
• /
• v.42 no.5
• /
• pp.277-280
• /
• 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.

• Asian Pacific Journal of Cancer Prevention
• /
• v.15 no.17
• /
• pp.7245-7249
• /
• 2014

Glucose regulated protein 78 (GRP78) is usually recognized as a chaperone in the endoplasmic reticulum. However, increasing evidence indicates that GRP78 can be translocated to the cell surface, acting as a signaling receptor for a variety of ligands. Since little is known about the secretion of GRP78 and its role in the progression of colon cancer we here focused on GRP78 from colon cancer cells, and purified GRP78 protein mimicking the secreted GRP78 was able to utilize cell surface GRP78 as its receptor, activating downstream PI3K/Akt and Wnt/${\beta}$-catenin signaling and promote colon cancer cell proliferation. Our study revealed a new mode of action of autocrine GRP78 in cancer progression: secreted GRP78 binds to cell surface GRP78 as its receptor and activates intracellular proliferation signaling.

• The Journal of Internal Korean Medicine
• /
• v.31 no.2
• /
• pp.273-289
• /
• 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.

• Nutrition Research and Practice
• /
• v.16 no.2
• /
• pp.161-172
• /
• 2022

BACKGROUND/OBJECTIVES: Colorectal cancer (CRC) is the third most common cancer worldwide and has a high recurrence rate, which is associated with cancer stem cells (CSCs). β-carotene (BC) possesses antioxidant activity and several anticancer mechanisms. However, no investigation has examined its effect on colon cancer stemness. MATERIALS/METHODS: CD133⁺CD44⁺ HCT116 and CD133⁺CD44⁺ HT-29 cells were isolated and analyzed their self-renewal capacity by clonogenic and sphere formation assays. Expressions of several CSCs markers and Wnt/β-catenin signaling were examined. In addition, CD133⁺CD44⁺ HCT116 cells were subcutaneously injected in xenograft mice and analyzed the effect of BC on tumor formation, tumor volume, and CSCs markers in tumors. RESULTS: BC inhibited self-renewal capacity and CSC markers, including CD44, CD133, ALDH1A1, NOTCH1, Sox2, and β-catenin in vitro. The effects of BC on CSC markers were confirmed in primary cells isolated from human CRC tumors. BC supplementation decreased the number and size of tumors and delayed the tumor-onset time in xenograft mice injected with CD133⁺CD44⁺ HCT116 cells. The inhibitory effect of BC on CSC markers and the Wnt/β-catenin signaling pathway in tumors was confirmed in vivo as well. CONCLUSIONS: These results suggest that BC may be a potential therapeutic agent for colon cancer by targeting colon CSCs.

• The Korean Journal of Food And Nutrition
• /
• v.35 no.5
• /
• pp.359-368
• /
• 2022

In this study, the quality characteristics of kimchi, such as its salinity, pH, and acidity, were measured and compared, and the HT-29 human colon cancer cells were used to show the anticancer effects of kimchi. The kimchi samples used herein included standard kimchi (SK), turnip kimchi (TK), and turnip-powder-added kimchi (TPK). The measured pH and acidity of TK and TPK showed no significant differences with those of SK. Compared to SK and TK, TPK had higher DPPH scavenging activity and higher total flavonoid content, confirming its antioxidant activity. The cancer cell growth inhibition rates of TK and TPK were significantly higher than that of SK. In HT-29 cells treated with TPK, the mRNA expression of Bcl-xL, an anti-apoptosis-related gene, was lower, and the mRNA expressions of the apoptosis-related genes Bax, Bad, and caspase-9 were higher. TPK showed significantly higher levels of mRNA expressions for the cell-cycle-related genes p53 and p21 than the other samples, in addition to suppression effects on cancer cell proliferation. Compared to SK, TK and TPK suppressed the growth of colon cancer cells and showed higher anticancer effects. Therefore, it is shown that kimchi with added turnip powder had high anticancer effects.

• Asian Pacific Journal of Cancer Prevention
• /
• v.15 no.8
• /
• pp.3767-3772
• /
• 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.

• Preventive Nutrition and Food Science
• /
• v.2 no.2
• /
• pp.167-173
• /
• 1997

The inhibitory effects of kale juice on the growh and DNA incorporation of human cancer cells, using HT-29 colon cancer cells, MG-63 osteosarcoma cells, AGS gastric adenocarcinoma cells and K-562 leukemia cells, were studied. The growth of human cancer cells were inhibited in the presence of kale juice (10, 20 nd 40$\mu$l/ml) and the effects were the juice concentration- and incubation time-dependent up to 6 days. When 20$\mu$l/ml of kale juice was added to the media of HT-29, MG-63, AGS and K-562 cancer cells, the cell growth after 6 or 4 days of incubation was retarded by 83~95% of control group. Morphological changes of HT-29 colon cancer cells wre studied under inverted microscope. As the concentration of kale juice increased up to 20$\mu$l/ml, degree of cell aggregation was decreased. Moreover, the DNA incorporation o AGS gastric adenocarcinoma cells and MG-63 osteosarcoma cells which were labeled with [$^3$H] thymidine was significantly reduced after 2 days of incubation at 37$^{\circ}C$ with kale juice. Therefore, we concluded that kale juice strongly decreased the growth of various human cancer cells.

• Asian Pacific Journal of Cancer Prevention
• /
• v.13 no.9
• /
• pp.4751-4756
• /
• 2012

Colon cancer continues to be one of the most common cancers, and the importance and necessity of new therapies needs to be stressed. The most important proto-oncogen factors for colon cancer appear to be epidermal growth factor receptor, EGFR, and c-Src with high expression and activity leading to tumor growth and ultimately to colon cancer progression. Application of c-Src and EGFR antisense agents simultaneously should theoretically therefore have major benefit. In the present study, anti-EGFR and c-Src specific antisense oligodeoxynucleotides were combined in a formulation using PAMAM dendrimers as a carrier. Nano drug entry into cells was confirmed by flow cytometry and fluorescence microscopy imaging and real time PCR showed gene expression of c-Src and EGFR, as well as downstream STAT5 and MAPK-1 with the tumor suppressor gene P53 to all be downregulated. EGFR and c-Src protein expression was also reduced when assessed by western blotting techniques. The effect of the antisense oligonucleotide on HT29 cell proliferation was determined by MTT assay, reduction beijng observed after 48 hours. In summary, nano-drug, anti-EGFR and c-Src specific antisense oligodeoxynucleotides were effectively transferred into HT-29 cells and inhibited gene expression in target cells. Based on the results of this study it appears that the use of antisense EGFR and c-Src simultaneously might have a significant effect on colon cancer growth by down regulation of EGFR and its downstream genes.

• Asian Pacific Journal of Cancer Prevention
• /
• v.16 no.6
• /
• pp.2569-2574
• /
• 2015

Necroptosis, also known as "programmed necrosis", has emerged as a critical factor in a variety of pathological and physiological processes and is considered a cell type-specific tightly regulated process with mechanisms that may vary rather greatly due to the change of cell line. Here we used HT-29, a human colon cancer cell line, to establish a necroptosis model and elucidate associated mechanisms. We discovered that cobalt chloride, a reagent that could induce hypoxia-inducible $factor-1{\alpha}(HIF1{\alpha})$ expression and therefore mimic the hypoxic microenvironment of tumor tissue in some aspects induces necroptosis in HT-29 cells when caspase activity is compromised. On the other hand, apoptosis appears to be the predominant death form when caspases are functioning normally. HT-29 cells demonstrated significantly increased RIPK1, RIPK3 and MLKL expression in response to cobalt chloride plus z-VAD treatment, which was accompanied by drastically increased $IL1{\alpha}$ and IL6 expression, substantiating the notion that necrosis can induce profound immune reactions. The RIPK1 kinase inhibitor necrostatin-1 and the ROS scavenger NAC each could prevent necrosis in HT-29 cells and the efficiency was enhanced by combined treatment. Thus by building up a necroptosis model in human colon cancer cells, we uncovered that mechanically RIP kinases collaborate with ROS during necrosis promoted by cobalt chloride plus z-VAD, which leads to inflammation. Necroptosis may present a new target for therapeutic intervention in cancer cells that are resistant to apoptotic cell death.

• Korean Journal of Plant Resources
• /
• v.28 no.6
• /
• pp.682-689
• /
• 2015

Abeliophyllum distichum Nakai (A. distichum) has been reported to exert the inhibitory effect on angiotensin converting enzyme and aldose reductase. Recently, our group found that branch extracts of A. distichum (EAFAD-B) induce apoptosis through ATF3 activation in human colon cancer cells. However, anti-cancer reagents exert their activity through the regulation of various molecular targets. Therefore, the elucidation of potential mechanisms of EAFAD-B for anti-cancer activity may be necessary. To elucidate the potential mechanism of EAFAD-B for anti-cancer activity, we evaluated the regulation of cyclin D1 in human colon cancer cells. EAFAD-B decreased cellular accumulation of cyclin D1 protein. However, cyclin D1 mRNA was not changed by EAFAD-B. Inhibition of proteasomal degradation by MG132 attenuated EAFAD-B-mediated cyclin D1 downregulation and the half-life of cyclin D1 was decreased in the cells treated with EAFAD-B. In addition, EAFAD-B induced cyclin D1 phosphorylation at threonine-286 and the point mutation of threonine-286 to alanine attenuated EAFAD-B-mediated cyclin D1 proteasomal degradation. Inhibitions of both ERK1/2 by PD98059 and NF-κB by a selective inhibitor, BAY 11-7082 suppressed cyclin D1 downregulation by EAFAD-B. From these results, we suggest that EAFAD-B-mediated cyclin D1 downregulation may result from proteasomal degradation through its threonine-286 phosphorylation via ERK1/2-dependent NF-κB activation. The current study provides new mechanistic link between EAFAD-B and anti-cancer activity in human colon cancer cells.