• Proceedings of the Ginseng society Conference
• /
• 1998.06a
• /
• pp.224-230
• /
• 1998

This study showed the anti-invasive activity of ginseng components, panaxadiol (PD) and panamatrlol (PT) on the highly metastatic HT1080 human flbrosarcoma cell line. PD and PT reduced tumor cell invasion through a reconstitute basement membrane in the transwell chamber. A significant down regulation of MMP-9 by PD and PT was detected by northern blot analysis. However, MMP-2 was constantly expressed. Quantitative gelatin based zymography confirmed a marked reduced expression of MMP-9 but not MMP-2 in the treatment of PD and PT. Since the chemical structures of PD and PT are very similar to that of dexamethasone, a synthetic glucocorticoid, it was investigated whether PD and PT act through GR. Western blot analysis and immunocytochemistry showed that PD and PT increased the GR fraction in the nucleus. These results suggest that ursolic acid may induce repression of MMP-9 by stimulating the nuclear translocation of GR and hence inhibiting the activity of AP-1 to TPA-responsible element of MMP-9 promoter region. In conclusion, we suggest that CR-induced down-regulation of MMP-9 by PD and PT contributes to reduce the invasive capacity of HT 1080 cells.

• YAKHAK HOEJI
• /
• v.47 no.4
• /
• pp.234-238
• /
• 2003

Linarin is a main compound from Chrysanthemum zawadskii var, latilobum. However, the biological mechanisms of these activities are unclear. Because of this wide diversity of effects, it is believed that they may be exerted through pluripotent effectors of linarin. In our previous screening study, the effects of linarin on the mouse macrophages cell line, RAW 264.7 cells, were investigated. It was found that linarin could stimulate macrophages activation by the production of tumor necrosis factor (TNF). The linarin (6.25∼12.5 $\mu\textrm{g}$/mι) inhibited the production of NO in LPS-activated RAW 264.7 cells and linarin became an useful candidates for the development of new drug to treat endotoxemia and inflammation accompanied by the overproduction of NO. However, linarin-treated total lymphocyte showed cytotoxicity in a dose dependent manner between 20 $\mu\textrm{g}$/mι and 40 $\mu\textrm{g}$/mι. In this study, linarin derivative (acetylated linarin) was synthesized in order to obtain less-cytotoxicity of linarin and evaluated for their in vitro cytotoxic activity aganist mouse total lymphocyte. There was no cytotoxic activity in a dose dependent manner (20∼40 $\mu\textrm{g}$/mι) of acetylated linarin whereas linarin showed. The production of NO, however, was not the case by this modified linarin. The cell morphological change was not significantly changed in response to acetylated linarin alone and these effects were potentiated by the addition of LPS. These results suggest that acetylated linarin may be developed to be a promising new drug candidate without cytotoxicity on the basis of its activity of macrophage activation.

• Journal of the East Asian Society of Dietary Life
• /
• v.27 no.1
• /
• pp.1-8
• /
• 2017

This study was conducted to investigate alterations of seaweed composition upon Lactobacillus rhamnosus GG (LGG) fermentation as well as potential anti-inflammatory effects and mechanism (s) of water extracts and fermented water extracts of Laminaria japonica (LJ) and Hizikia fusiforme (HF) in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. Total polyphenol, total sugar, and reducing sugar contents were measured in LJ and HF water extracts before and after fermentation by LGG. Alterations of inflammatory cytokine levels in cell culture media were measured by ELISA, and levels of phosphorylation of c-jun NH2-terminalkinase (JNK) and extra cellular signal regulated kinase (ERK) were examined by Western blot analysis. LGG fermentation of LJ and HF altered total polyphenol and sugar contents in water extracts of LJ and HF. LPS-induced production of pro-inflammatory cytokines such as IL-6 and $TNF-{\alpha}$ was significantly reduced by HF-f compared to control in RAW264.7 cells. Consistent with reduction of anti-inflammatory cytokine, interleukin (IL)-6, and tumor necrosis factor $(TNF)-{\alpha}$ levels by HF-f, HF-f also significantly reduced phosphorylation of ERK and JNK in LPS-stimulated RAW264.7 cells. In addition, LJ-f and HF also significantly reduced phosphorylation of JNK and ERK induced by LPS in RAW264.7 cells. Overall, our result suggests that HF-f among the four tested seaweed extracts is the most potent anti-inflammatory agent, and its mechanism of action is partially mediated by reduction of JNK and ERK phosphorylation as well as IL-6 and $TNF-{\alpha}$ production in LPS-stimulated RAW264.7 cells.

• The Korea Journal of Herbology
• /
• v.35 no.4
• /
• pp.37-43
• /
• 2020

Objectives : Allium hookeri is a well-known traditional herbal remedy and its root used for treatment of inflammation and tumor. However, the mechanism of anti-inflammatory effect of Allium hookeri is still unknown. This study aims to examine the mechanism of anti-inflammatory effect of Allium hookeri on mouse macrophage cell line, RAW 264.7 cells. Methods : Anti-oxidant effect of water extract of Allium hookeri (WEAH) was measured by 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) assay. 3- (4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay was performed to determine the effect of WEAH on cell viability in RAW 264.7 cells. In addition, anti-inflammatory effect of WEAH was investigated in RAW 264.7 cells. Inflammation of RAW 264.7 cells induced by lipopolysarccharide (LPS) treatment and expression levels of inflammatory cytokine interleukin 1 β (IL-1β) and interleukin 6 (IL-6) gene were analyzed using quantitative reverse transcription PCR (qRT-PCR) analysis. Furthermore, the phosphorylation of inhibitor of nuclear factor kappa B (IκBα) after LPS treatment with WEAH-treated RAW 264.7 cells was confirmed by immunoblot analysis. Results : WEAH showed a strong anti-oxidant effect and no cytotoxicity to RAW 264.7 cells up to 2 mg/㎖ concentration. The LPS-induced mRNA expression levels of IL-1β and IL-6 were decreased by WEAH treatment. Furthermore, the LPS-induced phosphorylation of IκBα is attenuated by WEAH treatment. Conclusions : Through experimental demonstration of anti-oxidant and anti-inflammatory effects of WEAH, we suggest that Allium hookeri is a valuable material for prevention and treatment of various inflammatory diseases.

• YAKHAK HOEJI
• /
• v.42 no.1
• /
• pp.82-88
• /
• 1998

The effect of hepatoprotective agents and bile acids on tumor necrosis factor-alpha, (TNF-${\alpha}$) production in murine and human macrophage cell line (RAW264.7 and U937) was inve stigated. The hepatoprotective agents including silymarin and its major component, silybin, significantly inhibited TNF-alpha production in a concentration dependent manner ($IC_50$ of silybin=67.7${\mu}g$/ml (140.3${\mu}g$M)). In differentiated U937 cells, especially, silybin showed more effective inbitory activity ($IC_50$=35.1${\mu}g$g/ml (72.7${\mu}g$M)). These results suggest that silymarin and silybin may inhibit TNF-alpha production in the process of hepatic diseases in human. However, biphenyldimethyl dicarboxylate (DDB) was not effective. In the case of bile acids, chenodeoxycholic acid (CDCA) showed a concentration dependent inhibitory effect on TNF-alpha production ($IC_50$ of CDCA= 71.5${\mu}g$g/ml (182.1${\mu}g$M)). In contrast, glycine or taurine conjugated form (G-CDCA or T-CDCA) restored to the control level or significantly increased TNF-${\alpha}$ production. And also ursodeoxycholic acid (UDCA) and its conjugated forms (G-UDCA and T-UDCA) showed a variety of patterns on TNF-${\alpha}$ production by changes of functional groups and concentration. These results also indicate that bile acids may regulate TNF-${\alpha}$ production in normal hepatic function or disease conditions.

• Journal of Animal Reproduction and Biotechnology
• /
• v.37 no.4
• /
• pp.218-225
• /
• 2022

High levels of proinflammatory cytokines have been observed in obese pregnancies. Obesity during pregnancy may increase the risk of various pregnancyrelated complications, with pathogenesis resulting from excessive inflammation. Palmitic acid (PA) is a saturated fatty acid that circulates in high levels in obese women. In our previous study, we found that PA inhibited the proliferation of trophoblasts developing into the placenta, induced apoptosis, and regulated the number of cleaved halves derived from transfer RNAs (tRNAs). However, it is not known how the expression of tRNA-derived stress-induced RNAs (tiRNAs) changes in response to PA treatment at concentrations that induce inflammation in human trophoblasts. We selected concentrations that did not affect cell viability after dose-dependent treatment of HTR8/SVneo cells, a human trophoblast cell line. PA (200 μM) did not affect the expression of apoptotic proteins in HTR8/SVneo cells. PA significantly increased the expression of inflammatory cytokines including interleukin (IL)-1β, IL-6, IL-8, and tumor necrosis factor (TNF)-α. In addition, 200 μM PA significantly increased the expression of tiRNAs compared to 800 μM PA treatment. These results suggest that PA impairs placental development during early pregnancy by inducing an inflammatory response in human trophoblasts. In addition, this study provides a basis for further research on the association between PA-induced inflammation and tiRNA generation.

• Tuberculosis and Respiratory Diseases
• /
• v.45 no.3
• /
• pp.587-595
• /
• 1998

Background: Recombinant adenovirus hold promise as vectors to carry therapeutic genes for several reasons: 1) they can infect both dividing and non-dividing cells; 2) they have the ability to directly transduce tissues in vivo; 3) they can easily be produced in high titer; and 4) they have an established record of safety as vaccination material. However, one of the major limitation in the use of adenoviruses is that transgene expression is quite short because adenovirusees insert their DNA genome episomally rather than by chromosomal integration, and an immune response against the virus destroys cells expressing the therapeutic gene. Since sodium butyrate has been reported to induce adenovirus-mediated gene expression, we hypothesized that treatment of tumor cells, transduced with herpes simples virus thymidine kinase(HSVtk) gene using adenoviral vector, with butyrate could augment the effect of gene therapy. Methods: We transduced HSVtk gene, driven by the cytomegalovirus promoter, into REN cell line(human mesothelioma cell line). Before proceeding with the comparison of HSVtk/ganciclovir mediated bystander killing, we evaluated the effect of butyrate on the growth of tumor cells in order to rule out a potential antitumor effect of butyrate alone, and also on expression of HSVtk gene by Western blot analysis. Then we determined the effects of butyrate on bystander-mediated cell killing in vitro. Results: There was no inhibition of growth of cells exposed to butyrate for 24 hours at a concentration of 1.5mM/L. Toxic effects were seen when the concentration of butyrate was greater than 2.0mM/L. Gene expression was more stable and bystander effect was augmented by butyrate treatment of a concentration of 1.5mM/L. Conclusion: These results provide evidence that butyrate can augment the efficiency of cell killing with HSVtk/GCV system by inducing transgene expression and may thus by a promising new approach to improve responses in gene therapy using adenoviral vectors.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.27 no.4
• /
• pp.314-320
• /
• 2001

Matrix metalloproteinases have long been viewed as ideal candidates for proteinases that enables tumor cells to permeated basement membrane defenses and invade surrounding tissue. There is growing evidence that the MMPs have an expanded role, as they are important for the creation and maintenance of a microenvironment that facilitates growth and angiogenesis of tumors at primary and metastatic sites. MT-MMPs are not secreted but instead remaining attached to cell surfaces. Although not all of the MT-MMPs are fully characterized, MT-MMPs have important role in localizing and activating secreted MMPs. The MMP genes are transcriptionally responsive to a wide variety of oncogene, growth factors, cytokine, and hormones. Currently, a number of MMP inhibitors are being developed and some have reached clinical trials as anti-metastatic or anti-cancer therapies. MT1-MMP is involved in the activation of proMMP-2. MT1-MMP is significant not only as a tumor marker but as a new target for chemotherapy against cancer. The purpose of this study was to evaluate the effects of protein kinase C inhibitor(genistein) on the proliferation of HT1080 and expression of MT1-MMP mRNA. Human fibrosarcoma cell line HT1080 was cultured and divided 2 groups. The experimental group was treated with $100{\mu}M$ genistein and incubated 12h, 24h for $[3^H]-thymidine$ uptake assay and northern hybridization individually. And the control group was treated with same amount of PBS for the above procedures. $[3^H]-thymidine$ incorporation was measured with ${\beta}$ ray detector. And RT-PCR and northern blotting for MT1-MMP mRNA was performed. The results were as follows 1. $[3^H]-thymidine$ uptake was reduced in experimental group with statistical significance. 2. MT1-MMP mRNA expression was significantly reduced in experimental group. These results showed that protein kinase C inhibitor (genistein) inhibited proliferation of HT1080 and almost completely blocked transcription of MT1-MMP mRNA. So, it is possible to use the protein kinase inhibitor (genistein) as anti-metastatic and anti-proliferative agent.

• Journal of Nutrition and Health
• /
• v.23 no.2
• /
• pp.135-147
• /
• 1990

This study was devised to observe the cytotoxic activities of garlic extracts against various cancer cells, that is, murine leukemic lymphocyte(L1210 and P388) and human rectal(HRT-18) and colon cancer cells(HCT-48 and HT-29) in vitro, and murine ascitic tumor cell(S-180) in vivo. Each cell-line except S-180 was cultured in medium containing serial concentration of the garlic extract in vitro. Inhibitory effect n the growth rate of the cancer cells was stronger in extracts of petroleum ether than that of ethanol. A lipid soluble compound in the extracts of garlic was cytocidal to murine leukemic cells, human rectal and colon cancer cells in vitro. The growth rates of the cancer cells in medium containing garlic extracts were inhibited gradually to a significant degree in proportion to the increase of the extract concentration. The cytotoxic activity of garlic active fraction from TLC was about 2.3 times more potent than that of crude garlic extract, one unit of cytotoxic activity against L1210 cells being equivalent to 4.2$\mu\textrm{g}$ and 1.8$\mu\textrm{g}$ from the crude garlic and active fraction, respectively. The Rf value of the active fraction on silica-gel TLC was 0.18 in condition that petroleum ether/ethyl ether/acetic acid mixture(90:10:1, v/v/v) was used as a developing solvent. The survival times of mice inoculated with S-180 cells were extended about 1.5 to 2 times in the groups treated with garlic extract(through i.p. and oral administration) compared with their control group(no garlic extract treatment). Observations were carried out on S-180. Ethanol extracts of garlic injured markedly tumor cells within 3 hours after injection.

• Asian Pacific Journal of Cancer Prevention
• /
• v.16 no.9
• /
• pp.4065-4069
• /
• 2015

Background: Epidermal growth factor-like domain multiple 7 (EGFL7), a secreted protein specifically expressed by endothelial cells during embryogenesis, recently was identified as a critical gene in tumor metastasis. Epithelial-mesenchymal transition (EMT) was found to be closely related with tumor progression. Accordingly, it is important to investigate the migration and EMT change after knock-down of EGFL7 gene expression in human pancreatic cancer cells. Materials and Methods: EGFL7 expression was firstly testified in 4 pancreatic cancer cell lines by real-time polymerase chain reaction (Real-time PCR) and western blot, and the highest expression of EGFL7 was found in PANC-1 cell line. Then, PANC-1 cells transfected with small interference RNA (siRNA) of EGFL7 using plasmid vector were named si-PANC-1, while transfected with negative control plasmid vector were called NC-PANC-1. Transwell assay was used to analyze the migration of PANC-1 cells. Real-time PCR and western blotting were used to detect the expression change of EGFL7 gene, EMT markers like E-Cadherin, N-Cadherin, Vimentin, Fibronectin and transcription factors like snail, slug in PANC-1, NCPANC-1, and si-PANC-1 cells, respectively. Results: After successful plasmid transfection, EGFL7 gene were dramatically knock-down by RNA interference in si-PANC-1 group. Meanwhile, migration ability decreased significantly, compared with PANC-1 and NC-PANC-1 group. Meanwhile, the expression of epithelial phenotype marker E-Cadherin increased and that of mesenchymal phenotype markers N-Cadherin, Vimentin, Fibronectin dramatically decreased in si-PANC-1 group, indicating a reversion of EMT. Also, transcription factors snail and slug decreased significantly after RNA interference. Conclusions: Current study suggested that highly-expressed EGFL7 promotes migration of PANC-1 cells and acts through transcription factors snail and slug to induce EMT, and further study is needed to confirm this issue.