• Journal of Marine Bioscience and Biotechnology
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• v.14 no.2
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• pp.93-101
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• 2022

Hizikia fusiforme, a type of brown algae, is widely used in Asian cuisine. It has been reported to have various pharmacological effects. In this study, the effects of the ethanol extract from H. fusiforme (EAHF) on the proliferation of human colon carcinoma cells were investigated. The effect on the survival of human hepatocarcinoma and colon carcinoma cells was examined, and results revealed that the anti-proliferative effects of EAHF were higher in colon carcinoma cells than in hepatocarcinoma cells. The inhibition of proliferation of HT29 colon carcinoma cells by EAHF treatment was closely related to the induction of apoptosis. EAHF treatment also increased caspase activity and poly(ADP-ribose) polymerase degradation, induced mitochondrial dysfunction, altered Bcl-2 family protein expression, and increased the rate of cytochrome c released from the mitochondria into the cytoplasm. Furthermore, the production of reactive oxygen species (ROS) was markedly stimulated by EAHF treatment, and when ROS production was blocked, EAHF-induced cytotoxicity was significantly attenuated. These results indicate that the anticancer activity of EAHF in HT29 colon carcinoma cells was induced by ROS-dependent mitochondrial impairment. While EAHF exhibited potent anticancer activity in colon carcinoma cells in this study, further studies on the active components of EAHF and their efficacy should be performed.

• Archives of Pharmacal Research
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• v.27 no.12
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• pp.1253-1257
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• 2004

6-Methoxydihydrosanguinarine (6ME), a benzophenanthridine alkaloid derived from the methanol extracts of Hylomecon hylomeconoides, showed a dose-dependent effect at 1-10 ${\mu}M$ on causing apoptotic cell death in HT29 colon carcinoma cells $(IC_{50} = 5.0{\pm}0.2 {\mu}M)$. Treatment of HT-29 cells with 6ME resulted in the formation of internucleosomal DNA fragmentation. Treatment of the cells with 6ME caused activation of caspase-3, -8 and 9 protease and subsequent proteolytic cleavage of poly(ADP-ribose)polymerase. 6ME increased the expression of p53 and Bax and decreased the expression of Bid. These results indicate that p53 and proapoptotic Bcl-2 family proteins might participate in the antiproliferative activity of 6ME in HT29 cells.

• Korean Journal of Food Science and Technology
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• v.23 no.3
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• pp.370-374
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• 1991

The inhibitory effects of garlic on the mutagenicity in Salmonella assay system and on the growth of HT-29 human colon carcinoma cells were studied. Methanol extract of garlic inhibited the mutagenicities induced by aflatoxin $B_1(AFB_1)$and N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) in Salmonella typhimurium TA100. The inhibition rate increased significantly when the concentration of the methanol extract from garlic increased in both strains of Salmonella typhimurium TA98 and TA100. The chloroform fraction from the methanol extract exhibited strong antimutagenicity against $AFB_1$. The chloroform fraction also inhibited greatly the growth of human HT-29 colon carcinoma cells in fetal bovine serum concentrations of 1% and 5%.

• Food Science and Biotechnology
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• v.16 no.2
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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.

• The Korean Journal of Physiology and Pharmacology
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• v.3 no.1
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• pp.75-82
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• 1999

It has been reported that activation of sphingomyelin pathway and nonsteroidal anti-inflammatory drugs (NSAIDS) inhibit the promotion of colon carcinoma. Ceramide, a metabolite of sphingomyelin, and indomethacin were shown to induce apoptosis in colon carcinoma cells. However, the mechanisms of ceramide- and indomethacin-induced apoptosis in the colon carcinoma cells are not clearly elucidated. Recent studys showed that indomethacin-induced apoptosis in colon cancer cells through the cyclooxygenase-independent pathways, and that may be mediated by generation of ceramide. In this study, we compared effects of ceramide and indomethacin on important modulators of apoptotic processes in HT29 cells, a human colon cancer cell line. Ceramide and indomethacin induced apoptosis dose- and time- dependently. Ceramide and indomethacin increased stress-activated protein kinase (SAPK) activity, and decreased mitogen-activated protein kinase (MAPK) activity. The expression of Bak was increased by the treatment of ceramide and indomethacin. The expression of other Bcl-2 related proteins (Mcl-1, $Bcl-X_L,$ Bax) which were known to be expressed in colon epithelial cells was not changed during the ceramide- and indomethacin-induced apoptosis. Our results suggest that ceramide and indomethacin share common mechanisms for induction of apoptosis in HT29 cells.

• Journal of Nutrition and Health
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• v.33 no.6
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• pp.660-667
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• 2000

Previous studies on the effect of age on intestinal taurine transport in animals have invariably shown a decline in the activity of the transport system with increasing age. In the present study changes in taurine transporter activity were observed during cell differentiation in the human colon carcinoma cell line HT-29 This cell line exhibits various enterocytic characteristics when differentiated and therefore has frequently been used to study the characteristcs and regulation of nutrient and drug absorption in the small intestine at the cellular level. Pre-treatment of the cells with $\beta$-alanine(10mM) reduced the taurine transport activity to 33% of the value for the control cells(p<0.05) which implies that taurine and $\beta$-alanine share a common $\beta$-amino acid transport system for their celluar uptake in the HI-29 was continued until 21 days post seeding. Kinetic studies of the taurine transporter were conducted in the HT-29 cell line with varying taurine concentration(5-60$\mu$M) in the uptake medium Both Vmax and the Michaelis-Menten constant(Km) of taurine transporter were decreased as differentiation of the HT-29 cell line was progressed ; Vmax of the taurine transporter in cells incubated for 4, 14 and 21 days post seeding was 2.79$\pm$3.4m 16.89$\pm$1.74, and 0.85$\pm$0.08 and 0.32$\pm$0.01nmol.mg protein-1 .30min-1 respectively(p<0.001) and Km was 42.3$\pm$3.4, 16.89$\pm$1.74, and 11.2$\pm$3.0$\mu$M respectively (p<0.01) These results indicate that the activity of sodium dependent active taurine transport system in the HT-29 cell line is decreased as confluent cells are differentiated. This phenomenon in cell culture system corresponds well with the earlier observation of lower intestinal taurine transport activity in suckling rats compared to that in adult animals although direct relationship of cell differentiation with in vivo aging process needs further verification.

• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.2
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• pp.314-319
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• 2001

In the present study, effects of $\beta$-alanine, a known taurine antagonist for its structural similarity, on the adaptive regulation and kinetic behavior of the taurine transporter were investigated in the HT-29, human colon carcinoma cell line. Pretreatment of the cell with $\beta$-alanine(10mM) for varying periods from 3 to 30 hrs significantly reduced the taurine uptake compared to the value for control cells. This decrease in the taurine transporter activity was dependent on the incubation time with $\beta$-alanine, and the maximal down-regulation of the transporter activity was observed in cells pretreated with $\beta$-alanine for 24 hrs (25% of the control value, p<0.01). The taurine transporter appears to bind exclusively with $\beta$-alanine in the HT-29 cells since the same concentration of $\alpha$-alanine added in the culture medium for 24 hrs did not influence the taurine uptake. Kinetic analyses of the taurine transporter activity was performed in the HT-29 cell line with varying taurine concentration (5~60$\mu$M) in the uptake medium. Active taurine uptake was significantly lower in $\beta$-alanine pretreated cells compared to the value for control cells in the range of taurine concentration used in the experiment (p<0.001). The cells pretreated with $\beta$-alanine showed a 50% lower maximal velocity (Vmax, 1.7$\pm$2.0 nmole.mg $protein^{-1}$.$30min^{-1}$), and a 99% higher Michaelis constant (Km, 40.3$\pm$7.6$\mu$M) than the control values (3.3$\pm$1.9 nmole.mg $protein^{-1}$.$30min^{-1}$, and 20.3$\pm$2.1$\mu$M, respectively). These results on kinetic data suggest that $\beta$-alanine induced down-regulation of the taurine transporter activity was associated with decreases in both maximal velocity and affinity of the transporter.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.12
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• pp.6017-6022
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• 2012

Background: Resveratrol has been reported to have potential chemopreventive and apoptosis-inducing properties in a variety of tumor cell lines. Objective: In this study, to investigate the effects of resveratrol on protein kinase C (PKC) activity and apoptosis in human colon carcinoma cells, we used HT-29 cells and examined the $PKC{\alpha}$ and ERK1/2 signaling pathways. Methods: To test the effects of resveratrol on the growth of HT-29 cells, the cells were exposed to varying concentrations and assessed with the the MTT cell-viability assay. Fluorescence-activated cell sorter (FACS) analysis was applieded to determine the effects of resveratrol on cell apoptosis. Western blotting was performed to determine the protein levels of $PKC{\alpha}$ and ERK1/2. In inhibition experiments, HT-29 cells were treated with G$\ddot{o}$6976 or PD98059 for 30 min, followed by exposure to $200{\mu}M$ resveratrol for 72 h. Results: Resveratrol had a significant inhibitory effect on HT-29 cell growth. FACS revealed that resveratrol induced apoptosis. Western blotting showed that e phosphorylation of $PKC{\alpha}$ and ERK1/2 was significantly increased in response to resveratrol treatment. Pre-treatment with $PKC{\alpha}$ and ERK1/2 inhibitors (G$\ddot{o}$6976 and PD98059) promoted apoptosis. Conclusion: Resveratrol has significant anti-proliferative effects on the colon cancer cell line HT-29. The PKC-ERK1/2 signaling pathway can partially mediate resveratrol-induced apoptosis of HT-29 cells.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.12
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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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• v.34 no.2
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• pp.150-157
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• 2001

Intestinal absorption of dietary taurine is one of the regulatory component maintaining taurine homeostasis along with renal reabsorption, bile acid conjugation and secretion, and de nobo synthesis of taurine in mammals. Recent observations of decreased enterocytic levels of taurine in response to trauma, infection and surgical insults, postulate the possibility that intestinal taurine absorption might be impaired in such stressed conditions. The aim of the present study was to evaluate changes in enterocytic taurine transporter activity using the human intestinal colon carcinoma cell line, HT-29, in various stress-induced conditions. Pretreatment of the HT-29 cells with dexamethasone, a stress hormone(0.1,1,10 or 100$\mu$M) for 3 hrs, or with E coli heat-stable enterotoxin(10, 100, or 200nM) for 30 minutes in order to induce the condition of enterotoxigenic infection did not influence taurine uptake as compared to the value found in control cells. In contrast, pretreatment of the cells with cholera toxin(10, 100, 500, or 1000ng/ml)for 3hr or 24hr significantly decreased taurine uptake by HT-29 cells to 40~50% of the value found in untreated control cells. Kinetic studies of the taurine transporter activity were conducted in control and cholera toxin treated HT-29 cells with varying taurine concentrations(2~60$\mu$M) in the uptake medium. Pretreatment of the cells with cholera toxin(100ng/ml) for 3hr did not influence the Vmax, but resulted in a 55% increase in the Michaelis-Menten constant(Km) of the taurine transporter compared to those in control cells. These results suggest that cholera toxin-induced reduction in taurine transporter activity in HT-29 cells is associated with decreased affinity of the taurine transporter without altering the amount of transporter protein. Intestinal taurine absorption appears to be reduced in the condition of water-borne diseases caused by bacteria such as V. cholerae. This might influence the taurine status of infants and young children more readily, an age group in which the prevalence of intestinal infection is high and the role of intestinal absorption is crucial for maintaining the body taurine pool. (Korean J Nutrition 34(2) : 150-157, 2001)