• Title/Summary/Keyword: HT-29 human

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Inhibitory Effect of Persimmon Leaves on the Mutagenicity in Spore Rec Assay and on the Growth of Human Cancer Cells (감잎의 용매별 추출물의 돌연변이 유발 억제 및 암세포 증식억제 효과)

  • 문숙희
    • The Korean Journal of Food And Nutrition
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    • v.15 no.1
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    • pp.23-28
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    • 2002
  • The inhibitory effects of persimmon leaves on th e mutagenicity in spore rec assay and on the growth of HT-29 human colon cancer cells and AZ-521 human gastric cancer cells were studied. Methanol extract of persimmon leaves inhibited the mutagenicity induced fly N-methyl- N'-nitro-N-nitrosoguanidine(MNNG) in spore rec assay. The hexane, chloroform and ethylacetate fraction from the methanol extract exhibited strong antimutagenicity against MNNG in spore rec assay The methanol extract of persimmon leaves also revealed the inhibitory effects on the growth of HT-29 human colon cancer cells and AZ-521 human gastric cancer cells. Among the solvent extracted fraction from the methanol extract, the chloroform fraction was most effective and inhibited the growth of HT-29 and AZ-521 cells by 100 percent.

Antimutagenic and Anticancer Effects of Leaf Mustard and Leaf Mustard Kimchi

  • Kim, Yong-Taek;Kim, Boh-Kyung;Park, Kun-Young
    • Preventive Nutrition and Food Science
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    • v.12 no.2
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    • pp.84-88
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    • 2007
  • In this study, we investigated antimutagenic and anticancer activities of leaf mustard (LM, Brassica juncea) and leaf mustard kimchi (LMK) during their fermentation period. Methanol extracts were prepared from raw mustard, brined leaf mustard in 10% Gueun salt solution for 2 hrs, leaf mustard fermented at 15$^{\circ}C$ for 5 days after brined in 10% Guenun salt solution for 2 hrs (Fr-LM), fresh leaf mustard kimchi (Fresh-LMK) and optimally ripened leaf mustard kimchi fermented at 5$^{\circ}C$ for 30 days (OR-LMK). OR-LMK showed the strongest inhibitory activities against the mutagenicities induced by aflatoxin B1 in Salmonella Typhimurium TA100. LMs and LMKs inhibited the survival or growth of AGS human gastric adenocarcinoma cells and HT-29 human colon carcinoma cells in MTT assay and growth inhibition test. Among the extracts, OR-LMK and FR-LM exhibited strong antiproliferative effect against cancer cells, especially HT-29 cells. DAPI staining assay showed that OR-LMK induced apoptosis cell death of HT-29 cells in a dose-dependent manner. These results suggest that leaf mustards and leaf mustard kimchi have chemopreventive activities.

Anticancer Effect of Doenjang in in vitro Sulforhodamine B (SRB) Assay (된장의 in vitro Sulforhodamine B (SRB) Assay에 의한 암세포 증식 억제 효과)

  • 이숙희;임선영;박건영
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.28 no.1
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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.

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사람의 Serine palmitoryl transferase II 및 ceramidase의 promoter에 대한 연구

  • Kim, Hui-Suk;Song, Seong-Gwang;Lee, Eun-Yeol;Lee, Sang-Do;Linn, Steve;Merrill, Alfred H.
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.588-591
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    • 2000
  • Serine palmitoyl transferase(SPT) and ceramidase are the key enzymes in sphingolipid biosynthesis. To study sphingolipid metabolism, we have got the 5'-upstream regions of human serine palmitoyl transferase subunit II and acid ceramidase gene by using GenomeWalker kits(Clontech Co.). Human genomic DNA was purified from HT29, human colon canser cell line by using DNAzol. We got several bands after secondary PCR and subcloned them to T7bule vector. Human SPTII promoter which we got was 2690bp but we cut it with Bgl II and vector with Bgl II and BamH I, and subcloned 1782bp to pGL2-enhancer vector and pGL2-basic vector with luciferase reporter gene. Human acid ceramidase promoter which we got were 2028bp and 1034bp and subcloned to pGL2-enhancer vector and pGL2-basic vector. We transfected these promoters to HT29 cell and assayed luciferase activity. For measuring transfection efficiency, pRL-TK vector with seapancy luciferase reproter gene was cotransfected with these promoters.

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$\beta$-Alanine Induced Down-Regulation of the Taurine Transporter Activity in the Human Colon Carcinoma Cell Line (HT-29) (인체 소장상피세포주 모델(HT-29)에서 $\beta$-알라닌이 타우린수송체 활성에 미치는 영향)

  • 박태선;윤미영;정한나;이해미
    • 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.

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Induction of Apoptosis in HT-29 Human Colorectal Cancer by Aloin (인간 대장암 세포 HT-29에서 Aloin에 의한 Apoptosis 유도)

  • Yoo, Eun-Seon;Woo, Joong-Seok;Kim, Sung-Hyun;Lee, Jae-Han;Han, So-Hee;Jung, Soo-Hyun;Park, Young-Seok;Kim, Byeong-Soo;Kim, Sang-Ki;Park, Byung-Kwon;Jung, Ji-Youn
    • Journal of Food Hygiene and Safety
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    • v.34 no.5
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    • pp.495-501
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    • 2019
  • Aloin [1,8-Dihydroxy-10-(${\beta}$-D-glucopyranosyl)-3-(hydroxymethyl)-9(10H)-anthracenone], is a natural anthraquinone from aloe. It has been shown to have antioxidant and anticancer effects in various types of human cancer cells, but the anticancer effects of aloin in human colorectal cancer cells HT-29 have not been elucidated. In this study, possible mechanisms by which aloin exerts its apoptotic action in cultured human colorectal cancer HT-29 cells were investigated. The results of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay shows that treatment with aloin (0, 100, 200, 300 and $400{\mu}M$) reduced cell viability in a concentration-dependent manner in HT-29 and showed no effects on cell proliferation in A375SM and AGS cells. In addition, it was confirmed that apoptotic body was significantly increased as shown by 4',6-diamidino-2-phenylindole (DAPI) staining, and increased apoptosis rate by flow cytometry in HT-29 cells treated with aloin (0, 200 and $400{\mu}M$). We confirmed by western blotting that aloin activated Bax (pro-apoptotic), cleaved-poly (ADP-ribose) polymerase (PARP) and caspase-3, -8 and Bcl-2 (anti-apoptotic) were not changed compared with the control. Aloin induced up-regulation of phospho-p38 and down-regulation of phospho-extracellular signal-regulated kinase (ERK)1/2. Therefore, aloin suppressed the growth inhibitory effects by the induction of apoptosis in human colorectal cancer cells and has potential as a cancer preventive medicine.

Resveratrol Affects Protein Kinase C Activity and Promotes Apoptosis in Human Colon Carcinoma Cells

  • Fang, Jie-Yu;Li, Zhi-Hua;Li, Qiang;Huang, Wen-Sheng;Kang, Liang;Wang, Jian-Ping
    • 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.

Cytotoxic and Apoptotic Activities of Tussilago farfara Extract in HT-29 Human Colon Cancer Cells

  • Lee, Mi-Ra;Cha, Mi-Ran;Jo, Kyung-Jin;Yoon, Mi-Young;Park, Hae-Ryong
    • Food Science and Biotechnology
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    • v.17 no.2
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    • pp.308-312
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    • 2008
  • The flower buds of Tussilago farfara (TF) have been traditionally used in oriental medicine for the treatment of bronchitis and asthma. In our study, the primary objective was to determine the mechanisms that are inherent to TF-induced cytotoxicity and apoptosis, using the methanolic extract of TF (TFM) in HT-29 human colon cancer cells. We found that TFM-induced induced cytotoxicity in HT-29 cells in a dose-dependent manner. This effect was verified via an MTT reduction assay, an lactate dehydrogenase (LDH) release assay, and a colony formation assay. Interestingly, we also detected apoptotic bodies on Hoechst staining, and attempted to determine whether TFM-induced apoptosis involved the caspase pathway using a caspase-3/7 activity assay. Overall, the results indicate that TFM contain chemotherapeutic agents and potential candidates use for against human colon cancer cells.

Induction of ROS-dependent apoptosis by ethanol extract of Hizikia fusiforme in HT29 colon carcinoma cells (톳 에탄올 추출물에 의한 HT29 결장암 세포의 ROS 의존적 세포사멸 유도)

  • Su Hyun, Hong;Yung Hyun, Choi
    • 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.

Antiproliferative Effect of Opuntia humifusa Ethanol Extract on Human Carcinoma HT-29 Cells (천년초 선인장 줄기 에탄올 추출물의 HT-29 대장암 세포증식 저해효과)

  • Park, Soo Young;Kim, Young A;Ly, Sun Yung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.12
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    • pp.1827-1834
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    • 2014
  • Colon cancer is the third highest cause of death in Korea. Known dietary causes of colon cancer include a diet rich in fat and red meat as well as inadequate intake of dietary fiber, fruits, and vegetables. Therefore, recent research has focused on the anticancer effects of natural products. Opuntia humifusa is a type of prickly pear that is known to contain biologically active compounds that can be used in the treatment of diabetes mellitus, arteriosclerosis, and hyperglycemia. The aim of this study was to determine whether or not O. humifusa extract affects proliferation, cell death, and DNA fragmentation in human carcinoma HT-29 cells. O. humifusa is rich in carbohydrates, minerals (Mg, K, and Ca), and total phenolics. HT-29 cells were treated with extracts of O. humifusa at concentrations of 0, 0.25, 0.5, 1, and 2 mg/mL for 24 or 48 hours. O. humifusa extracts inhibited HT-29 cell growth in a dose-dependent manner. Hoechst 33342/PI double staining and Comet assay were performed to observe changes in nuclei of cancer cells undergoing cell death. The results of both tests showed that O. humifusa extract induced cell shrinkage, DNA fragmentation, and chromatin condensation dose-dependently in HT-29 cells. The results of this study suggest that O. humifusa extract inhibits the growth of HT-29 via induction of DNA fragmentation and chromatin condensation.