• Title/Summary/Keyword: GT1-1 cells

Search Result 65, Processing Time 0.025 seconds

Inhibition of Sphingolipid Metabolism Enhances Resveratrol Chemotherapy in Human Gastric Cancer Cells

  • Shin, Kyong-Oh;Park, Nam-Young;Seo, Cho-Hee;Hong, Seon-Pyo;Oh, Ki-Wan;Hong, Jin-Tae;Han, Sang-Kil;Lee, Yong-Moon
    • Biomolecules & Therapeutics
    • /
    • v.20 no.5
    • /
    • pp.470-476
    • /
    • 2012
  • Resveratrol, a chemopreventive agent, is rapidly metabolized in the intestine and liver via glucuronidation. Thus, the pharmacokinetics of resveratrol limits its efficacy. To improve efficacy, the activity of resveratrol was investigated in the context of sphingolipid metabolism in human gastric cancer cells. Diverse sphingolipid metabolites, including dihydroceramides (DHCer), were tested for their ability to induce resveratrol cytotoxicity. Exposure to resveratrol ($100{\mu}M$) for 24 hr induced cell death and cell cycle arrest in gastric cancer cells. Exposure to the combination of resveratrol and dimethylsphingosine (DMS) increased cytotoxicity, demonstrating that sphingolipid metabolites intensify resveratrol activity. Specifically, DHCer accumulated in a resveratrol concentration-dependent manner in SNU-1 and HT-29 cells, but not in SNU-668 cells. LC-MS/MS analysis showed that specific DHCer species containing C24:0, C16:0, C24:1, and C22:0 fatty acids chain were increased by up to 30-fold by resveratrol, indicating that resveratrol may partially inhibit DHCer desaturase. Indeed, resveratrol mildly inhibited DHCer desaturase activity compared to the specific inhibitor GT-11 or to retinamide (4-HPR); however, in SNU-1 cells resveratrol alone exhibited a typical cell cycle arrest pattern, which GT-11 did not alter, indicating that inhibition of DHCer desaturase is not essential to the cytotoxicity induced by the combination of resveratrol and sphingolipid metabolites. Resveratrol-induced p53 expression strongly correlated with the enhancement of cytotoxicity observed upon combination of resveratrol with DMS or 4-HPR. Taken together, these results show that DHCer accumulation is a novel lipid biomarker of resveratrol-induced cytotoxicity in human gastric cancer cells.

Polymorphic Lengths of Dinucleotide $(GT)^n$ Repeats in Upstream of Human nNOS Exon 1f Gene Play a Role in Modulating the nNOS Transcription: Clinical Implications

  • Shin, Mi-Kyung;Kim, Kyung-Nam;Kim, Chul-Eung;Lee, Sung-Keun;Kang, Ju-Hee;Park, Chang-Shin
    • Molecular & Cellular Toxicology
    • /
    • v.4 no.1
    • /
    • pp.11-15
    • /
    • 2008
  • The expression of neuronal nitric oxide synthase (nNOS) is regulated by various spliced first exons (exon 1a-1i), sharing differentially common exon 2 in diverse human tissues. The highly complex structure and regulation of human nNOS gene gave limitations of information for the precise mechanism of nNOS regulation. In the present study, we report that the repeats of polymorphic dinucleotides $(GT)^nA(TG)^n$ repeats located in just upstream to the exon 1f in human nNOS gene play suppressive role in transcription, as shown in the characteristics of Z-DNA motif in other genes. In neuronal and trophoblast cells transfected transiently with luciferase construct without dinucleotide repeats at the 5'-flanking region of exon 1f in nNOS gene, the luciferase activity was increased markedly. However, the presence of the dinucleotide repeats dramatically suppressed the luciferase activity to the basal level, and which was dependent on the length of $(GT)^n$ and $(TG)^n$ repeats. More importantly, we found the polymorphisms in the length of dinucleotide repeats in human. Furthermore, we show for the first time here that there is a significant association of the lengths of polymorphic dinucleotide $(GT)^n$ and $(TG)^n$ repeats with the risk of schizophrenia.

Glycosyltransformation of ginsenoside Rh2 into two novel ginsenosides using recombinant glycosyltransferase from Lactobacillus rhamnosus and its in vitro applications

  • Wang, Dan-Dan;Kim, Yeon-Ju;Baek, Nam In;Mathiyalagan, Ramya;Wang, Chao;Jin, Yan;Xu, Xing Yue;Yang, Deok-Chun
    • Journal of Ginseng Research
    • /
    • v.45 no.1
    • /
    • pp.48-57
    • /
    • 2021
  • Background: Ginsenoside Rh2 is well known for many pharmacological activities, such as anticancer, antidiabetes, antiinflammatory, and antiobesity properties. Glycosyltransferases (GTs) are ubiquitous enzymes present in nature and are widely used for the synthesis of oligosaccharides, polysaccharides, glycoconjugates, and novel derivatives. We aimed to synthesize new ginsenosides from Rh2 using the recombinant GT enzyme and investigate its cytotoxicity with diverse cell lines. Methods: We have used a GT gene with 1,224-bp gene sequence cloned from Lactobacillus rhamnosus (LRGT) and then expressed in Escherichia coli BL21 (DE3). The recombinant GT protein was purified and demonstrated to transform Rh2 into two novel ginsenosides, and they were characterized by nuclear magnetic resonance (NMR) techniques and evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide assay. Results: Two novel ginsenosides with an additional glucopyranosyl (6→1) and two additional glucopyranosyl (6→1) linked with the C-3 position of the substrate Rh2 were synthesized, respectively. Cell viability assay in the lung cancer (A549) cell line showed that glucosyl ginsenoside Rh2 inhibited cell viability more potently than ginsenoside Rg3 and Rh2 at a concentration of 10 μM. Furthermore, glucosyl ginsenoside Rh2 did not exhibit any cytotoxic effect in murine macrophage cells (RAW264.7), mouse embryo fibroblasts cells (3T3-L1), and skin cells (B16BL6) at a concentration of 10 μM compared with ginsenoside Rh2 and Rg3. Conclusion: This is the first report on the synthesis of two novel ginsenosides, namely, glucosyl ginsenoside Rh2 and diglucosyl ginsenoside Rh2 from Rh2 by using recombinant GT isolated from L. rhamnosus. Moreover, diglucosyl ginsenoside Rh2 might be a new candidate for treatment of inflammation, obesity, and skin whiting, and especially for anticancer.

Antimicrobial Peptides from Lactobacillus plantarum UTNGt2 Prevent Harmful Bacteria Growth on Fresh Tomatoes

  • Tenea, Gabriela N.;Pozo, Tatiana Delgado
    • Journal of Microbiology and Biotechnology
    • /
    • v.29 no.10
    • /
    • pp.1553-1560
    • /
    • 2019
  • In a previous study, the antimicrobial peptides extracted from Lactobacillus plantarum UTNGt2 of wild-type fruits of Theobroma grandiflorum (Amazon) were characterized. This study aimed to investigate the antimicrobial mechanisms of peptides in vitro and its protective effect on fresh tomatoes. The addition of partially purified Gt2 peptides to the E. coli suspension cells at the exponential ($OD_{605}=0.7$) growth phase resulted in a decrease with 1.67 (log10) order of magnitude compared to the control without peptide. A marginal event (< 1 log10 difference) was recorded against Salmonella, while no effect was observed when combined with EDTA, suggesting that the presence of a chelating agent interfered with the antimicrobial activity. The Gt2 peptides disrupted the membrane of E. coli, causing the release of ${\beta}$-galactosidase and leakage of DNA/RNA molecules followed by cell death, revealing a bacteriolytic mode of action. The tomatoes fruits coated with Gt2 peptides showed growth inhibition of the artificially inoculated Salmonella cocktail, demonstrating their preservative potential.

Effects of the Feeds Supplemented with Korean Green Tea and Lactic Acid Bacteria on Infection of Eimeria maxima in Chickens (닭에서 녹차 및 유산균 혼합 사료 투여가 Eimeria maxima의 감염에 미치는 효과)

  • Jang, Seung-Ik;Chung, Nyun-Ki;Min, Won-Gi;You, Myung-Jo;Park, Bae-Keun;Jun, Moo-Hyung
    • Journal of Veterinary Clinics
    • /
    • v.23 no.3
    • /
    • pp.243-250
    • /
    • 2006
  • The chickens fed with the feeds supplemented with green tea(GT) and lactic acid bacteria(LB) were infected orally with 10,000 oocysts per chicken of E. maxima. The groups administered with the feeds supplemented with GT by 0.5% and 2.0% of feed showed the significant levels of decreasing in the number of oocysts shed for 5 days after E. maxima infection. The feeds supplemented with LB by 0.1% and 0.5% of feed were less effective in reducing the number of the fecal oocyst, compared with the groups administered with GT. To evaluate the immunoregulatory effects of the feed additives, the expression patterns of IL-2 and IFN-r in spleen cells were studied by RT-PCR and ELISA. The higher levels of IL-2 transcripts after E. maxima infection were observed in the groups with n, compared with the groups with LB and the mixture of GT and LB. The $IFN-\gamma$ mRNA bands were observed in the all of experimental groups except the uninfected control. The culture supernatants of Con A-stimulated spleen cells($5{\times}10^6cells/ml$) were measured for the concentration of IL-2 and $IFN-\gamma$ by ELISA. The levels of IL-2 and $IFN-\gamma$ on days 3 and 7 after E. maxima infection were significantly augmented in the groups with n. These results indicated that GT-supplemented feeds resulted in higher reduction of oocyst-shedding and more enhanced immune responses in the chicken infected with E. maxima, as compared with LB-supplemented feeds. According to the results, it was implicative that the supplements could be utilized for development of feed additives for anti-coccidiosis.

The Alleviating Effects of Green Tea on N-Nitrosodimethylamine Toxicity to Mucins in the Rat Lingual Salivary Glands (흰쥐 혀침샘의 점액질에 미치는 N-Nitrosodimethylamine 독성에 대한 녹차의 완화 효과)

  • Cho, Eun-Ju;Jeong, Gil-Nam;Jo, Un-Bock
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.37 no.12
    • /
    • pp.1568-1575
    • /
    • 2008
  • This study was performed to determine alleviating effects of green tea (GT) on the N-nitrosodimethylamine (NDMA) toxicity to mucins in the rat lingual salivary glands. Sprague-Dawley rats were divided into five groups: untreated (Control), one week-NDMA administrated (NDMA), one week-GT administrated after NDMA for one week (NDMAGT), one week-NDMA administrated with GT (GTNDMA), and one-week NDMA administrated with GT after GT for one week (GTGTNDMA). The mucin properties were elucidated by periodic acid Schiff (PAS) reaction, alcian blue (AB) pH 2.5, AB pH 2.5-PAS, aldehyde fuchsin (AF) pH 1.7-AB pH 2.5, and high iron diamine (HID)-AB pH 2.5 staining. The lingual von Ebner's glands of NDMA group showed some changes such as contraction and destruction of the serous acini, decrease and disappearance of the cytoplasmic granules, vacuolation of cytoplasm, and the mucigenous duct cells. Also, in the lingual mucous glands, enlarged lumens, fused acini, disappearance of granules, vacuolation of cytoplasm, and mucigenous duct cells were observed. All the GT administrated groups had a tendency to recover. GTGTNDMA group recovered almost up to the state of the control group. The lingual von Ebner's glands of NDMA group showed a decrease of neutral mucin and the lingual mucous glands showed a decrease of both neutral and acidic mucins decreased in comparison with control group, although mucous acini secreting strong sulfomucin decreased while those secreting non-sulfomucin (sialomucin) increased. All the GT administrated groups had a tendency to recover. The degree of recovery in the GTNDMA group was stronger than in NDMAGT group and that of the GTGTNDMA group was almost up to the state of the control group. Plus, lingual von Ebner's glands of GTGTNDMA group contained much more neutral mucin than in the control group. In conclusion, it is suggested that NDMA exhibit the toxicity which affects on the mucin properties and which green tea alleviates.