• Title/Summary/Keyword: AP-PCR

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Inhibitory Effect of Naringenin on MMP-9 Activity and Expression in HT-1080 Cells (Naringenin이 NF-$\kappa$B, AP-1 억제를 통한 MMP-9 활성 및 발현 억제 효과)

  • Chae, Soo-Chul;Kho, Eun-Gyeong;Seo, Eun-Sun;Ryu, Geun-Chang;Na, Myung-Suk;Kim, In-Suk;Lee, Jong-Bin
    • Korean Journal of Environmental Biology
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    • v.27 no.1
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    • pp.58-65
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    • 2009
  • The chemopreventive effects of naringenin derived from citrus on tumor migration and the possible mechanisms involved in this protection were investigated in HT-1080 tumor cells. In this study, we found that naringenin reduced phorbol 12-myristate 13-acetate (PMA)-enhanced matrix metalloproteinases (MMP)-9 activation in a dose-dependant manner and further inhibited HT-1080 cell migration. In addition, naringenin suppressed PMA-enhanced expression of MMP-9 protein, mRNA and transcription activity levels through suppression of nuclear factor $\kappa$B (NF-$\kappa$B) activation and activator protein-1 (AP-1) translocation without changing tissue inhibitor of metalloproteinase (TIMP)-1 level. Therefore, our results suggested that the inhibitory effects of naringenin on MMP-9 activation, relation of tumor migration in vitro possibly involve mechanisms related to its ability to suppress PMA-enhanced MMP-9 gene and protein expression through NF-$\kappa$B activation and AP-1 translocation. Overall, naringenin may be a valuable anti-invasive drug candidate for cancer therapy.

New Gene Profiling in Determination of Breast Cancer Recurrence and Prognosis in Iranian Women

  • Poorhosseini, Seyed Mohammad;Hashemi, Mohammad;Olyaei, Nasrin Alipour;Izadi, Amir;Moslemi, Elham;Ravesh, Zeinab;Hashemi-Gorji, Feyzollah;Kheiri, Hamid Reza;Yassaee, Vahid Reza
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.sup3
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    • pp.155-160
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    • 2016
  • Breast cancer (BC) is the second most common cancer in the world and by far the most frequent cancer among women, with an estimated 1.67 million new cancer cases diagnosed in 2012 (25% of all cancers). Polygene expression analysis is used to predict the prognosis and determine the most appropriate treatment regimen. The objective of this study was to examine the gene expression profiles of SIRT3, HRAS, LSP1, SCUBE2 and AP2A2 in Iranian women with BC.A total of 136 patients including healthy controls were categorized into three groups based on the relapse of the disease. Expression of desired genes in formalin-fixed, paraffin embedded tissues collected from all groups of participants was analyzed via the RT PCR method. RNA extraction and cDNA synthesis were performed then real-time quantitative PCR was carried out. Gene expression analysis revealed that the expression of SIRT3 was equal among patient and control groups. LSP1 was down regulated in all patient groups relative to controls but reduced expression in the metastatic group relative to the non-metastatic one was not significant. HRAS was significantly overexpressed in total and metastatic tumor samples versus normal but not in non-metastatic cases. SCUBE2 expression showed significant over-expression in both overall tumor samples and the non-metastatic group as compared to normal tissues. Gene expression level of AP2A2 in all groups was not detectable. Our data are compatible with a tumor suppressor role of LSP1 related to potential prognostic factor for tumor recurrence and outcome. This study for the first time assayed the prognostic value and changes in the expression of SIRT3, LSP1, HRAS, SCUBE2 and AP2A2 genes in women with breast cancer in the Iranian population and findings confirmed potential biomarker and prognostic capability of these genes. Such expression profiling data can critically improve prognosis and treatment decisions in cancer patients.

Anti-wrinkle Effect by Ginsenoside Rg3 Derived from Ginseng (인삼유래 Ginsenoside Rg3에 의한 항-주름 효과)

  • 김성우;정지헌;조병기
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.2
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    • pp.221-225
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    • 2004
  • The root of Panax ginseng C. A. Meyer has been used as a traditional anti-aging and anti-wrinkle agent in the Orient. However, it is still unknown which component of ginseng is effective at suppressing wrinkle formation. Recently at least twenty ginsenosides regarded as the main active ingredients of ginseng have been isolated. Among them, we examined the effect of ginsenoside Rg3 on dermal ECM metabolism to elucidate the mechanism of anti-wrinkle by ginseng. In our study, to investigate the anti-wrinkle effect of the ginsenoside Rg3, ECM component and growth factor in dennis were evaluated by ELISA assay. Ginsenoside Rg3 was found to stimulate type I procollagen and fibronectin (FN) biosynthesis in a dose-dependent manner in normal human fibroblast culture (p < 0.05, n =3), and dose-dependently enhance TGF- ${\beta}$1 level (p < 0.05, n =3). In RT-PCR analysis mRNA level of c-Jun, a member of AP-1 transcription factor, was reduced by ginsenoside Rg3 in normal human fibroblast culture. These results indicate that ginsenoside Rg3 stimulates type I collagen and FN synthesis through the changes of TGF - ${\beta}$1 and AP-1 expression in fibroblasts.

Isolation of labial Gene in the Oligochaeta Perionyx excavatus and Expression Patterns during Head Regeneration (지렁이 labial 유전자의 동정 및 머리재생 시 발현 양상에 관한 연구)

  • Cho Sung Jin;Lee Myung Sik;Tak Eun Sik;Hur So Young;Lee Jong Ae;Park Bum Joon;Cho Hyun Ju;Shin Chuog;Park Soon Cheol
    • The Korean Journal of Soil Zoology
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    • v.8 no.1_2
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    • pp.1-6
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    • 2003
  • Hox genes are pivotal in the control of morphogenesis along the anterior-posterior (AP) axis in various bilaterians. Several indications suggest their involvement in the control of cell growth and regeneration. For the labial full-length fragment, RACE-PCR was employed to obtain the 3' and 5' franking regions. Semi-quantitative RT-PCR analysis revealed that the labial expression began to increase at 12 hours after amputation. The peak expression was approximately 1.5-fold more than the unamputated controls. This result could give us information on the significance of Hox genes and the relationships between Hox genes during regeneration.

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Antimicrobial Resistance and Molecular Epidemiologic Characteristics of Stenotrophomonas maltophilia Isolated from Clinical Specimens (병원 재료에서 분리한 Stenotrophomonas maltophilia의 항균제 내성 및 분자역학적 특성)

  • Seol, Sung-Yong;Jang, Kyoung-Soo;Jeong, Oung-Gi;Cho, Eung-Rae;Kim, Neung-Hee;Yu, Hak-Sun;Lee, Yoo-Chul;Cho, Dong-Taek
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.3
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    • pp.239-250
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    • 2000
  • Sixty-eight clinical isolates of Stenotrophomonas maltophilia from inpatients of 2 university hospitals in Taegu were epidemiologically analyzed by using the minimum inhibitory concentrations of 25 antimicrobial drugs, biochemical reaction, pulsed-field gel elctropgoresis (PFGE), and PCR with enterobacterial repetitive intergenic consensus sequences as primer (ERIC-PCR). 1. All the strains were susceptible to minocycline. More than 57% were susceptible to sulfisomidine (Su), ciprofloxacin (Ci), Ofloploxacin (Of), nalidixic acid (Na), and chloramphenicol (Cm), and $19{\sim}35%$ to ceftazidime (Cd), trimethoprim (Tp), Ticacillin-clavulanic acid, and cefoperazone-sulbactam. Most isolates were resistant to ${\beta}$-lactam antibiotics such as ampicillin (Ap), carbenicillin (Cb), cefotaxim (Ct), cefoxitin (Cx), and aminoglycosides including gentamicin (Gm), tobramycin (Tb), amikacin (Ak). 2. All the isolates were multiply resistant of 5 to 17 drugs and showed 40 different resistance pattern types. 3. All the strains showed very similar biochemical reactions except ${\beta}$-galactosidase and nitrate reduction test. Fourteen strains selected randomly were classified 10 different pattern type by PFGE and ERIC-PCR. These two methods showed identical result. Four strains isolated from wound in 1994 showed similar MIC pattern and identical API 20NE profile, PFGE, and ERIC-PCR pattern indicating episodes of cross-infection among patients. These results indicate that PFGE or ERIC-PCR profile has comparable discriminatory power for epidemiological typing of S. maltophilia.

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Effect of Sulforaphane on LPS-Induced Matrix Metalloproteinase-9 (MMP-9) Expression (Sulfolaphane이 lipopolysaccharide (LPS)에 의해 유도된 matrix metalloproteinase-9 (MMP-9) 발현에 미치는 영향)

  • Lee, Jung-Tae;Woo, Kyung-Jin;Kwon, Taeg-Kyu
    • Journal of Life Science
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    • v.20 no.2
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    • pp.275-280
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    • 2010
  • Sulforaphane is a naturally occurring member of the iosothiocyanate family, which reveals chemopreventive capacities including anti-cancer, anti-inflammation and inhibition of MMP-9 activities. In this study, we investigated the effect of sulforaphane on the expression of matrix metalloproteinase-9 (MMP-9) in lipopolysaccharide (LPS)-induced Raw 264.7 cells. Sulforaphane strikingly suppressed the LPS-induced MMP-9 activity and mRNA expression in a dose-dependent manner. In addition, sulforaphane inhibited not only the LPS-induced MMP-9 promoter activity but also LPS-mediated activator protein-1 (AP-1) and nuclear factor-kB (NF-${\kappa}B$) promoter activity. Transient transfection by MMP-9 constructs, in which specific transcriptional factors were mutagenized, indicated that the effects of LPS and sulforaphane were mediated via AP-1 and NF-${\kappa}B$ response elements. We found that sulforaphane had the ability to suppress LPS-induced invasion in vitro. Taken together, these results demonstrated that sulforaphane effectively suppressed LPS-induced MMP-9 expression via modulation of promoter elements (AP-1 and NF-${\kappa}B$) in MMP-9 transcriptional activation.

Protective effects of Dictamni Radicis Cortex water extract on acute pancreatitis (백선피(白鮮皮) 물 추출물의 급성 췌장염 보호 효과)

  • Kim, Dong-Uk;Bae, Gi-Sang;Choi, Ji-Won;Kim, Dong-Gu;Kim, Myoung-Jin;Song, Ho-Joon;Park, Sung-Joo
    • The Korea Journal of Herbology
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    • v.34 no.1
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    • pp.75-80
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    • 2019
  • Objectives : Dictamni Radicis Cortex (DRC) has been used as an important traditional medicine for inflammation and fungal diseases. However, the protective effect of DRC water extract on acute pancreatitis (AP) has not been deeply reported. Therefore, we aimed to evaluate the protective effects of DRC water extract on cerulein-induced AP. Methods : AP was induced via intraperitoneal injection of supramaximal concentrations of stable cholecystokinin analogue cerulein ($50{\mu}g/kg$) every hour for 6 times. DRC water extract (0.05, 0.1, or 0.2 g/kg) or saline was administrated intraperitoneally 1 h before to the first injection of cerulein. The mice were sacrificed at 6 h after the final cerulein injection. Pancreas was rapidly removed for histochemical examination and myeloperoxidase (MPO) assay. In addition, polymerase chain reaction (PCR) was performed to examine mRNA levels of proinflammatory cytokines such as Interleukin $(IL)-1{\beta}$, IL-6 and Tumor necrosis factor $(TNF)-{\alpha}$. Results : Administration of DRC water extract significantly inhibited the pancreatic weight to body weight ratio, pancreas histological damages and increase of pancreatic MPO activity during cerulein-induced AP. In addition, increased pancreatic mRNA levels of $IL-1{\beta}$, IL-6 but not $TNF-{\alpha}$ were significantly inhibited by treatment of DRC water extract against cerulein-induced AP. Conclusions : In conclusion, we have revealed that pre-treatment of DRC water extract reduces the severity of cerulein-induced AP. Accordingly, our results could give a clinical basis that DRC could be used as a drug or agent to prevent AP.

γ-Glutamyltranspeptidase Gene from Bacillus subtilis BS 62 (Bacillus subtilis BS 62의 γ-Glutamyltranspeptidase 유전자)

  • Lee, Tae-Eun;Yoon, Min-Ho;Choi, Woo-Young
    • Korean Journal of Agricultural Science
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    • v.34 no.2
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    • pp.161-170
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    • 2007
  • To characterize $\gamma$-glutamyltranspeptidase ($\gamma$-GTP or ggt; EC 2. 3. 2. 2.) gene of Bacillus subtilis BS 62, the $\gamma$-GTP gene of BS 62 was prepared from PCR products amplified with the chromosomal DNA. The $\gamma$-GTP gene of about 2.5 kb was sequenced, and its homology was compared with the other ggt genes which were reported previously. The base sequence of the gene appeared to have an open reading frame of 1,758 bp encoding a protein of 62,175 Da. The coding region was flanked by putative ribosome binding site - AGGAGG of 7th to 12th upstream - and the stem-loof sequence was followed by transcription terminator codon. Homology of the amino acid residues sequence consisting of 587 amino acid residues was found as 98% with Bacillus subtilis gene (BSU49358), 97.4% with that of Bacillus subtilis KX 102, 37% with Pseudomonas sp. A14 (S63255) and 38% with Streptomyces avermitils (AP005028).

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Berberine suppresses in vitro migration of human aortic smooth muscle cells through the inhibitions of MMP-2/9, u-PA, AP-1, and NF-κB

  • Liu, Su-Jian;Yin, Cai-Xia;Ding, Ming-Chao;Xia, Shao-You;Shen, Qin-Min;Wu, Ji-Dong
    • BMB Reports
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    • v.47 no.7
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    • pp.388-392
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    • 2014
  • Berberine, a type of isoquinoline alkaloid isolated from Chinese medicinal herbs, has been reported to have various pharmacological activities. Studies have demonstrated that berberine has beneficial effects on vascular remodeling and alleviates restenosis after vascular injury. However, its mechanism of action on vascular smooth muscle cell migration is not fully understood. We therefore investigated the effect of berberine on human aortic smooth muscle cell (HASMC) migration. Boyden chamber assay was performed to show that berberine inhibited HASMC migration dose-dependently. Real-time PCR and Western blotting analyses showed that levels of matrix metalloproteinase (MMP)-2, MMP-9, and urokinase-type plasminogen activator (u-PA) were reduced by berberine at both the mRNA and protein levels. Western blotting assay further confirmed that activities of c-Fos, c-Jun, and NF-${\kappa}B$ were significantly attenuated. These results suggest that berberine effectively inhibited HASMC migration, possibly by down-regulating MMP-2, MMP-9, and u-PA; and interrupting AP-1 and NF-${\kappa}B$ mediated signaling pathways.

Cloning And Characterization of Pathogen-Inducible EREBP-Like Transcription Factor(CaNR19) From Hot Pepper (Capsicum annuum L.)

  • Yi, So-Young;Kim, Jee-Hyub;Yu, Seung-Hun;Park, Doil
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.77.2-78
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    • 2003
  • An EREBP/AP2-type transcription factor (CaPFl) was isolated by DDRT-PCR following inoculation of soybean pustule pathogen Xanthomonas axonopodis pv. glycines Bra which induces HR on pepper leaves. Genomic Southern blot analysis revealed that the CaPFl gene is present as a single copy within the hot pepper genome. The deduced amino acid sequence of CaPFl has two potential nuclear localization signals, a possible acidic activation domain, and an EREBP/AP2 motif that could bind to a conserved cis- element present in promoter region of many stress-induced genes. The mRNA level of CaPFl was induced by both biotic and abiotic stresses. We observed higher-level transcripts in resistance-induced pepper tissues than diseased tissues. Expression of CaPFl is also induced upon various abiotic stresses including ethephon, MeJA, cold stress, drought stress and salt stress treatments. To study the role of CPFI in plant, transgenic Arabidopsis and tobacco plants which express higher level of pepper CaPFl were generated. Global gene expression analysis of transgenic Arabidopsis by cDNA microarray indicated that expression of CaPFl in transgenic plants affect the expression of quite a few GCC box and DRE/CRT box-containing genes. Furthermore, the transgenic Arabidopsis and tobacco plant, expressing CaPFl showed tolerance against freezing temperature and enhanced resistance to Pseudomonas syrnigae pv. tabaci. Taken together, these results indicated that CaPFl is a novel EREBP/AP2 transcription factor in hot pepper plant and it may has a significant role(s) in regulation of biotic and abiotic stresses in plant.

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