• Animal Bioscience
• /
• 제36권10호
• /
• pp.1604-1611
• /
• 2023

Objective: The aim of this study was to investigate the protective effect of wheat phytase as a structural decomposer of inflammatory nucleotides, extracellular adenosine triphosphate (ATP), and uridine diphosphate (UDP) on HT-29 cells. Methods: Phosphatase activities of wheat phytase against ATP and UDP was investigated in the presence or absence of inhibitors such as L-phenylalanine and L-homoarginine using a Pi Color Lock gold phosphate detection kit. Viability of HT-29 cells exposed to intact- or dephosphorylated-nucleotides was analyzed with an EZ-CYTOX kit. Secretion levels of pro-inflammatory cytokines (IL-6 and IL-8) in HT-29 cells exposed to substrate treated with or without wheat phytase were measured with enzyme-linked immunosorbent assay kits. Activation of caspase-3 in HT-29 cells treated with intact ATP or dephosphorylated-ATP was investigated using a colorimetric assay kit. Results: Wheat phytase dephosphorylated both nucleotides, ATP and UDP, in a dose-dependent manner. Regardless of the presence or absence of enzyme inhibitors (L-phenylalanine and L-homoarginine), wheat phytase dephosphorylated UDP. Only L-phenylalanine inhibited the dephosphorylation of ATP by wheat phytase. However, the level of inhibition was less than 10%. Wheat phytase significantly enhanced the viability of HT-29 cells against ATP- and UDP-induced cytotoxicity. Interleukin (IL)-8 released from HT-29 cells with nucleotides dephosphorylated by wheat phytase was higher than that released from HT-29 cells with intact nucleotides. Moreover, the release of IL-6 was strongly induced from HT-29 cells with UDP dephosphorylated by wheat phytase. HT-29 cells with ATP degraded by wheat phytase showed significantly (13%) lower activity of caspase-3 than HT-29 cells with intact ATP. Conclusion: Wheat phytase can be a candidate for veterinary medicine to prevent cell death in animals. In this context, wheat phytase beyond its nutritional aspects might be a novel and promising tool for promoting growth and function of intestinal epithelial cells under luminal ATP and UDP surge in the gut.

• Journal of Pharmaceutical Investigation
• /
• 제25권2호
• /
• pp.137-143
• /
• 1995

The peptide transporter can be utilized for improving the bioavailability of compounds that are poorly absorbed. Characterization of the dipeptide uptake into the human intestinal epithelial cells, HT-29 was investigated. The uptake of tritiated glycylsarcosine $([^3H]-Gly-Sar,\;0.1\;{\mu}Ci/ml)$ was measured in confluent or subconfluent HT-29, Caco-2, and Cos-7 cells. Uptake medium was the Dulbecco's Modified Eagle's Media (DMEM) adjusted to pH 6.0. Both HT-29 and Caco-2 cells expressed the dipeptide transporter significantly (p<0.005) but Cos-7 did not. Certain portions of passive uptake were observed in all three cell lines. Uptake of Gly-Sar was largest at 7 days after plating HT-29 cells with significant inhibition with 25 mM cold Gly-Sar (p<0.05). but expression ratio of the dipeptide transporter was 0.7, suggesting lower expression. The effect of pH on Gly-Sar uptake was not significant in the range of pH 6 to 8. Gly-Sar uptake was also inhibited with 50 mM carnosine, 25 mM Gly-Sar, and 35 mM cephalexin significantly (p<0.05). From above results the dipeptide transporter was expressed well in HT-29 cells and was similar to that in the small intestine, suggesting that large amounts of mRNA of the transporter from the cells can be obtained.

• 동아시아식생활학회지
• /
• 제24권1호
• /
• pp.20-27
• /
• 2014

Our study is investigated the effects of Nelumbo nucifera root extract on HT-29 colon cancer cells. The anti-proliferative effect of 70% ethanol extract from Nelumbo nucifera root on HT-29 colon cancer cells was identified based on cell viability, Hoechst 33342 nuclear staining, apoptosis analysis, Western blotting and RT-PCR analyses. In our study, Nelumbo nucifera root extract inhibited the growth of HT-29 colon cancer cells in a dose-dependent manner. Concomitant activation of the mitochondria-dependent apoptotic pathway of HT-29 colon cancer cells by Nelumbo nucifera root extract occurred via modulation of Bax and Bcl-2 expressions, which activated cleavage of caspases-3 and -9. The findings of this study indicate that Nelumbo nucifera root extract induces apoptosis in HT-29 colon cancer cells, and this phenomenon is occurs via the death receptor-mediated and mitochondria-mediated apoptotic pathways.

• 한방안이비인후피부과학회지
• /
• 제24권1호
• /
• pp.16-24
• /
• 2011

Objective : In this study, we investigated the effects of ethanol extract of Anemarrhenae Rhizoma (EAR) on the proliferation and apoptosis induction of HT-29 human colon cancer cells. Methods : Cell viability of HT-29 cells were measured by MTT assay and apoptisis-related proteins were assessed using western blotting. Chromatin condensation of HT-29 cells stained with Hoechst 33258. Results : In the present study, we demonstrated that EAR exhibited significant cytotoxicity in HT-29 cells. The induction of apoptosis in HT-29 cells by EAR treatment was characterized by chromatin condensation and the activation of caspase-3. EAR-induced apoptosis is accompanied by the release of cytochrome c and the specific proteolytic cleavage of PARP. EAR was appeared cytotoxic effect to HT-29 cells in a dose-dependent manner. Concomitantly, EAR treatment led to increase in the caspase-9. The reduction of Bcl-2 and truncation of Bid were induced by EAR. Conclusion : We studied that the EAR induced apoptosis in human colon adenocarcinoma HT-29 cells. These results indicated that EAR can cause apoptosis through mitochondria/caspase pathway in human HT-29 cells.

• 생약학회지
• /
• 제42권3호
• /
• pp.282-287
• /
• 2011

This study was presented to the anti-cancer activity from different fraction of Zanthoxylum schnifolium fruits. The values for human colon cancer cell(HT-29) survival rate of 0.3 mg/mL of 70% EtOH and 70% MeOH ethyl acetate fraction extracts were 7.62${\pm}$0.173%, 7.66${\pm}$0.037%, respectively. It was shown that human colon cancer cell(HT-29) survival rate was in a dose-dependent manner. The percentages of cells were increased in the sub-G0 and G0/G1 phase region, meaning that cell proliferation was decreased. The RT-PCR demonstrated that 70% EtOH and 70% MeOH ethyl acetate fraction extracts were down-regulated the expression of Bcl-2 and survivin genes in HT-29 cells. We examined that 70% EtOH and 70% MeOH ethyl acetate fraction extracts anti-cancer activities initiated through ROS generation suggesting that HT-29 cells treated with ethyl acetate fraction extracts induced ROS generation. Our results revealed that the Zanthoxylum schnifolium fruit may expect for anti-cancer activities in HT-29 cells.

• Parasites, Hosts and Diseases
• /
• 제51권1호
• /
• pp.61-68
• /
• 2013

Entamoeba histolytica, which causes amoebic colitis and occasionally liver abscess in humans, is able to induce host cell death. However, signaling mechanisms of colon cell death induced by E. histolytica are not fully elucidated. In this study, we investigated the signaling role of NOX in cell death of HT29 colonic epithelial cells induced by E. histolytica. Incubation of HT29 cells with amoebic trophozoites resulted in DNA fragmentation that is a hallmark of apoptotic cell death. In addition, E. histolytica generate intracellular reactive oxygen species (ROS) in a contact-dependent manner. Inhibition of intracellular ROS level with treatment with DPI, an inhibitor of NADPH oxidases (NOXs), decreased Entamoebainduced ROS generation and cell death in HT29 cells. However, pan-caspase inhibitor did not affect E. histolytica-induced HT29 cell death. In HT29 cells, catalytic subunit NOX1 and regulatory subunit Rac1 for NOX1 activation were highly expressed. We next investigated whether NADPH oxidase 1 (NOX1)-derived ROS is closely associated with HT29 cell death induced by E. histolytica. Suppression of Rac1 by siRNA significantly inhibited Entamoeba-induced cell death. Moreover, knockdown of NOX1 by siRNA, effectively inhibited E. histolytica-triggered DNA fragmentation in HT29 cells. These results suggest that NOX1-derived ROS is required for apoptotic cell death in HT29 colon epithelial cells induced by E. histolytica.

• Food Science and Biotechnology
• /
• 제14권6호
• /
• pp.866-870
• /
• 2005

The 14 lactic acid bacteria (LAB) have been evaluated to determine the binding capacity to HT29 cell and Aflatoxin $B_1$ ($AFB_1$). The interaction of LAB to HT29 cells has been further investigated to identify the possibility of competing the binding sites with $AFB_1$. Of 14 LAB strains, Lactobacillus casei KCTC 3260 demonstrated the higher adhesiveness to HT29 and $AFB_1$ with the rate of 19.6% and 46.3%, respectively. In competitive analysis for binding sites, the adhesion of L. casei KCTC 3260 to HT29 cells was reduced with 100 nmol $AFB_1$ by 31.2%. The protoplast of L. casei KCTC 3260 showed no binding capacity to HT29 cells with increment of $AFB_1$ concentration, indicating that cell wall components might serve as a critical factor for the binding. To discriminate the major component influencing on L. casei KCTC 3260 binding to HT29 cells and $AFB_1$, four different pre-treatments (lipase, pronase E, sodium m-periodate, and urea) were employed. Of those, sodium m-periodate treatment caused the lower adhesion of L. casei KCTC 3260 to HT29 cells with the increment of $AFB_1$ concentration. These results indicated that carbohydrate moiety on the cell wall of L. casei KCTC 3260 might be the most critical component in binding to both HT29 cells and $AFB_1$.

• 대한본초학회지
• /
• 제22권2호
• /
• pp.65-72
• /
• 2007

Objectives : In this study, we investigate that Euphorbiae lathyridis Semen extract contributes to growth inhibitory effect and anti-cancer activity on the HT-29 human colon cancer cells. Methods : Euphorbiae lathyridis Semen was extracted from the Semen of the plant using 80% Methanol. The Euphorbiae lathyridis Semen extract was treated to different concentrations for 24 hr, 4Shr or 72hr. Growth inhibitory effect was analyzed by measuring FACS study and MTT assay. Cell apoptosis was confirmed by surveying caspases cascades activation using Westem blot. Results : Exposure to Euphorbiae lathyridis Semen extract (0.4mg/ml) results in an inhibitory effect on cell growth in HT-29 cells. Growth inhibition by Euphorbiae lathyridis Semen extract in HT-29 cells was related with the inhibition of proliferation and induction of apoptosis. The Euphorbiae lathyridis Semen extract induces DNA fragmentation in HT-29 cells. Furthermore, Euphorbiae lathyridis Semen extract induces cell apoptosis through the activation of caspases-3, caspase-9 and PARP cleavage. Conclusion : Euphorbiae lathyridis Semen extract induces apoptosis in human colon cancer cells, therefore, we suggest that Euphorbiae lathyridis Semen extract can be used as a novel class of anti-cancer drugs.

• 동의생리병리학회지
• /
• 제26권2호
• /
• pp.155-159
• /
• 2012

The purpose of this study was to investigate the anti-cancer effects of Sophorae Radix and the effects of 5-Fluorouracil (5-FU) in human colorectal adenocarcinoma cells (HT-29). We used human colorectal adenocarcinoma cell line, HT-29 cells. We examined cell death by MTT assay and caspase 3 assay with Sophorae Radix. To examine the inhibitory effects of Sophorae Radix, cell cycle (sub G1) analysis was done the HT-29 cells after three days with Sophorae Radix. The reversibility of Sophorae Radix was examined on one day to five days treatment with $150{\mu}g$ Sophorae Radix. Sophorae Radix inhibited the growth of HT-29 cells in a dose-dependent fashion. Also we showed that Sophorae Radix induced apoptosis in HT-29 cells by MTT assay, caspase 3 assay and sub-G1 analysis. Sophorae Radix combined with 5-FU markedly inhibited the growth of HT-29 cells compared to Sophorae Radix or 5-FU alone. After 3 days treatment of HT-29 cells with Sophorae Radix, the fraction of cells in sub-G1 phase was much higher than that of the control group. Our findings provide insight into unraveling the effects of Sophorae Radix in human colorectal adenocarcinoma cells and developing therapeutic agents against colorectal cancer.

• Journal of Nutrition and Health
• /
• 제34권2호
• /
• pp.150-157
• /
• 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)