• IMMUNE NETWORK
• /
• v.1 no.2
• /
• pp.151-161
• /
• 2001

Background: Inactivation in p53 tumor suppressor gene through a point mutation and deletion is one of the most frequent genetic changes found in human cancer, with 50% of an incidence. This high rate of mutation mostly suggests that the gene plays a central role in the development of cancer and the mutations detected so far were found in exons 5 to 8. Mutation of p53 locus produced accumulation of abnormal p53 protein, and negative regulation of cell proliferation and transcriptional activation as a suppressor of transformation were lost. In addition, inhibition of its normal cellular function of wild-type by mutant is an important step in tumorigenesis. Method: 4 colon cancer cell lines (SNU C1, C2A, C4, C5) were examined for mutation in exons 5 to 8 of the p53 tumor suppressor gene by PCR-SSCP analysis and expression pattern by western blotting and immunoprecipitation. p53-mediated transactivation ability were examined by CAT assay and base substitution of p53 in SNU C2A cell were detected by DNA sequencing. Results: 1) SNU C2A cell and SNU C5 cell were detected mobility shifts each in exon 5 and exon 7 of p53 gene by the PCR-SSCP method, implicating being of p53 mutation. 2) 3 colon cancer cell lines (SNU C1, SNU C2A, SNU C5) expressed wild type and mutant type p53 protein. 3) In northern blot experiment, SNU C2A and SNU C5 cell expressed high level of p53 mRNA. 4) Results of p53-mediated transactivation in colon cancer cell lines by CAT assay represented only SNU C2A cell has transcriptional activity. 5) DNA sequencing in SNU C2A cell showed missense mutation in codon 179 of one allele, histidine to arginine and wild type p53 in the other allele. Conclusion: Colon cancer cell lines showed correlation with mutation in p53 gene and accumulation of abnormal p53 protein. Colon cancer cell SNU C2A retained p53-mediated transactivation as heterozygous p53 with one mutant allele in 179 codon and the other wild-type allele.

• The Korean Journal of Physiology and Pharmacology
• /
• v.5 no.6
• /
• pp.521-527
• /
• 2001

Diclofenac, a phenylacetic acid derivative, is a widely used non-steroidal anti-inflammatory drug (NSAID) to provide effective relief of inflammation and pain. Nitric oxide (NO) synthesized by inducible nitric oxide synthase (iNOS) has been implicated as a mediator of inflammation. We examined the inhibitory effects of diclofenac on the induction of iNOS in RAW 264.7 macrophages which were activated with lipopolysaccharide (LPS) plus interferon-gamma $(IFN-{\gamma}).$ Treatment of RAW 264.7 cells with diclofenac and other NSAIDs (aspirin and indomethacin) significantly inhibited NO production and iNOS protein expression induced by LPS plus $IFN-{\gamma}.$ Also, diclofenac but not aspirin and indomethacin, inhibited iNOS mRNA expression and nuclear factor-kappa B $(NF-{\kappa}B)$ binding activity concentration-dependently. Furthermore, transfection of RAW 264.7 cells with iNOS promoter linked to a CAT reporter gene revealed that only diclofenac inhibited the iNOS promoter activity induced by LPS plus $IFN-{\gamma}$ through the $NF-{\kappa}B$ sites of iNOS promoter. Taken together, these suggest that diclofenac may exert its anti-inflammatory effect by inhibiting iNOS gene expression at the transcriptional level through suppression of $NF-{\kappa}B$ activation.

• Mycobiology
• /
• v.46 no.4
• /
• pp.382-387
• /
• 2018

The entomopathogenic fungus Cordyceps militaris is a valuable medicinal ascomycete, which degenerates frequently during subsequent culture. To avoid economic losses during industrialized production, scratching stimuli of mycelia was introduced to improve the fruiting body production. The present results indicated that higher yields and biological efficiency were obtained from two degenerate strains (YN1-14 and YN2-7) but not from g38 (an insertional mutant in Rhf1 gene with higher yields and shorter growth periods). Furthermore, the growth periods of the fruiting bodies were at least 5 days earlier when the mycelia were scratched before stromata differentiation. Three ROS-scavenging genes including Cu/Zn superoxide dismutase (CmSod1), Glutathione peroxidase (CmGpx), and Catalase A (CmCat A) were isolated and their expression profiles against scratching were determined in degenerate strain YN1-14 and mutant strain g38. At day 5 after scratching, the expression level of CmGpx significantly decreased for strain g38, but that of CmSod1 significantly increased for YN1-14. These results indicated that scratching is an effective way to promote fruiting body production of degenerate strain, which may be related at least with Rhf1 and active oxygen scavenging genes.

• Microbiology and Biotechnology Letters
• /
• v.17 no.6
• /
• pp.600-605
• /
• 1989

For the purpose of isolation of the Zymomonas mobilis DNA fragments showing promoter activity in Escherichia coli, a promoter screening vector, PCMT215 was constructed by transferring a promoterless chloramphenicol acetyltransferase (CAT) gene of pYEJ001 into pMT21 which contains $\beta$-lactamase gene and multiple cloning sites. A library of Z, mobilis Sau3AI DNA fragments was constructed in E. coli using the newly constructed pCMT215. Fourteen clones showing resistance to chloramphenicol ranging in concentration from 30 to 750 $\mu$g/$m\ell$ were selected. From five clones of them, the Z. mobilis DNA fragments expressing CAT gene of the recombinant plasmids were sequenced and then sites of transcriptional initiation were identified. The nucleotide sequences of the cloned DNA shared AT rich regions, poly A's or T's stretches and palindromic regions. The positions of transcriptional initiation for CAT gene occurred at more than one site spaced over by 4 to 190 base pairs on the cloned fragments in E. coli.

• YAKHAK HOEJI
• /
• v.50 no.1
• /
• pp.58-63
• /
• 2006

The antioxidant enzyme (AOE) system plays an important role in the defense against oxidative stress damage. To determine whether myricetin or myricetin/taurine can exert antioxidative effects not only by modulating the AOE system directly but also by scavenging free radical, we investigated the influence of the myricetin and taurine on cell viability ROS level, activities of different antioxidant enzyme, and the expression of different antioxidant enzyme. As results, the cell viability showed inhibition of the proliferation with treatment of 'myricetin' or 'myricetin with taruine', respectively, with dose-dependent manner. Compared to control, the treatment of 'myricetin' decreased activities and gene expressions of superoxide dismutase (SOD), and glutathione peroxidase (GPx). However, combined treatment of 'myricetin with taurine' increased activities and gene expressions of the SOD, GPx, and catalase (CAT). In addition, the combined treatment of 'myricetin with taurine' somewhat decreased ROS levels, compared to the treatment of 'myricetin'. In conclusion, our study provides that the combined treatment of different antioxidants can enhance antioxidant effects.

• Bulletin of the Korean Chemical Society
• /
• v.26 no.9
• /
• pp.1385-1389
• /
• 2005

Human hepatitis B virus (HBV) is a pathogen related to the development of liver diseases including chronic hepatitis, cirrhosis, and hepatocellular carcinoma (HCC). However, the efficient methods to suppress HBV replication have not been developed yet. Therefore, we have used RNA interference (RNAi) as a potential tool for the suppression of HBV replication. Here, we designed a 21 nt small intefering dsRNA (siRNA) against hepatitis B virus X (HBx) RNA with 3' overhanging ends derived from T7 promoter. It has been reported that HBV X protein plays an important role in HBV gene expression and viral replication. The suppression of HBx gene expression by the 21 nt siRNA was investigated by Northern blot analysis and chloramphenicol acetyl transferase (CAT) assay. The level of HBx mRNA was decreased by siRNA in a dose-dependent manner. We also found that the 21 nt siRNA inhibited the HBV replication in hepatocellular carcinoma cell.

• Korean Journal of Plant Resources
• /
• v.24 no.1
• /
• pp.55-60
• /
• 2011

To investigate effects of Ixeris dentata EtOH ext. on lowering lipid levels and anti-oxidation activity, hyperlipidemic rats were treated with Ixeris dentata EtOH ext. and lipid levels and anti-oxdation activity were measured. The Ixeris dentata EtOH ext. groups showed low concentration of plasma FFA, plasma triglyceride, plasma total cholesterol, and plasma LDL-cholesterol compared to control group. However, concentration of plasma HDLcholesterol was not significantly different among all the treatment groups. The Ixeris dentata EtOH ext. groups showed lower level of liver total cholesterol, liver triglyceride, plasma TBARS, and liver TBARS than those of control group. The Ixeris dentata EtOH ext. groups also showed higher level of GSH-Px activity, SOD activity, and CAT activity than those of control group. Moreover, the Ext. showed lower level of TNF-${\alpha}$, Apo-B, Apo-E, and leptin expression than those of control group. The results in this study shows that the Ixeris dentata EtOH ext. have positive effect in lowering lipid level, and anti-oxidative activity.

• Journal of Aquaculture
• /
• v.19 no.3
• /
• pp.157-165
• /
• 2006

Expression of major antioxidant enzyme (AOE) including Cu/Zn superoxide dismutase (Cu/Zn-SOD), catalase (CAT), glutathione-S-transferase (GST) and 3 glutathione peroxidase isotypes (GPXs) at mRNA levels during heat stress was examined in mud loach (Misgurnus mizolepis) liver. Based on the semi-quantitative RT-PCR, real-time RT-PCR and/or northern dot blot hybridization, the antioxidant enzyme genes were generally up-regulated during elevation of water temperature from $23^{\circ}C$ up to $32^{\circ}C$. GPXs and SOD displayed the most significant elevation of mRNA levels (up to 3 and 2 folds, respectively) while CAT showed the steady-state expression irrespective of thermal conditions. GST represented the relatively moderate response (1.3-fold increase) in its transcription to thermal stress. The transcriptional activation of AOE genes was not significant at the treatment temperature lower than $29^{\circ}C$. Increased mRNA levels of GPX (extracellular form) and SOD genes in the fish exposed to $32^{\circ}C$ was readily detectable 1 day after exposure to heat stress.

• Journal of Veterinary Clinics
• /
• v.37 no.4
• /
• pp.208-212
• /
• 2020

A 6-year-old castrated male Russian Blue cat was presented for evaluation of dyschezia. Abdominal ultrasound revealed hyperechoic nodules in both kidneys, heterogeneous mass in abdomen, and extensive mesenteric thickening with multiple hypoechoic nodules. Computed tomography showed multiple hypodense lesions in both kidneys and diffuse nodular infiltration around the mesentery. Fine needle aspirates (FNA) acquired under ultrasound guidance from the mesentery consisted of large lymphocytes which have round to irregular nuclei with granular chromatin, prominent nucleoli and a small amount of basophilic cytoplasm. Polymerase chain reaction (PCR) for antigen receptor gene rearrangement result of FNA sample revealed a T-cell malignancy. The cat died from acute renal failure after 1 cycle of modified Madison-Wisconsin L-CHOP protocol. Postmortem examination revealed bilaterally enlarged lumpy-bumpy shaped kidneys. Histopathologic examination showed an infiltration of malignant lymphocytes into the renal parenchyma and mesentery. Immunohistochemical staining of the renal sample displayed a negative expression of CD3, PAX5, MUM-1, and CD79. The clinical features and prognosis of the cat with renal lymphoma with mesenteric lymphomatosis have been described in this report.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.28 no.5
• /
• pp.520-529
• /
• 2014

The purpose of this study is to investigate the protective effects of Yukmijiwhang-tang(YM) water extracts against the UVB irradiation on the human keratinocyte HaCaT cells. We observed the effects of YM on the oxidative stress, gene expression of pro-inflammatory cytokine such as TNF-${\alpha}$ and IL-$1{\beta}$, and matrix metalloproteinase-9 in UVB-irradiated HaCaT cells. On the effects of oxidative stress and antioxidant function on the treatment with YM, The activity of xanthine oxidase(XO) was significantly decreased by treatment of YM in all the concentrations(p<0.01). The activity of superoxide dismutase(SOD) and catalase(CAT) was significantly increased by treatment of YM in a dose dependent manner(p<0.05 and p<0.01). DPPH radical was erased by treatment of YM under dose of $500{\mu}g/m{\ell}$ concentration. Treatment of HaCaT cells with YM had also significantly reduced intracellular ROS produced by UVB irradiation in a dose dependent manner(p<0.05, p<0.01, p<0.001). Gelatin zymography assay showed that YM downregulated the MMP-9 activity in UVB-irradiated HaCaT cells. RT-PCR analysis revealed that YM suppressed the expression of IL-$1{\beta}$ and MMP-9 however, it has no effects on the expression of TNF-${\alpha}$ and MMP-3. Our study suggests that Yukmijiwhang-tang exert protective actions on the UVB-irradiated HaCaT cells largely by anti-oxidative and anti-inflammatory processes.