• Toxicological Research
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• 제25권1호
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• pp.35-40
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• 2009

TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin) and related halogenated aromatic hydrocarbons elicit a diverse spectrum of biochemical and toxic responses in laboratory animals and mammalian cells in culture. Toxicity and carcinogenicity of TCDD is well established but the molecular mechanism is still poorly understood. Here, we found the noble responsive genes to TCDD using the differential display analysis. Treatment of HepG2 cells with TCDD showed a significantly different mRNA expression pattern from the untreated cells in differential display analysis. The differentially displayed bands were isolated and used as probes in dot blot and Northern blot analyses. Of thirty-five isolated differentially displayed bands, only two bands were confirmed as positive in dot blot and Northern blot analyses. The nucleotides sequences of these clones were analyzed and the search of Genebank database revealed that one clone is highly homologous with RanBP2 (Ras-related nuclear protein binding protein2; 92%) and the other is an unknown gene. RanBP2 is a nucleoporin with SUMO E3 ligase activity that functions in both nucleocytoplasmic transport and mitosis and its role as a novel tumor suppressor has been recently proposed. Thus, these results may suggest the clue elucidating the toxic mechanism of TCDD through RanBP2.

• Natural Product Sciences
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• 제12권4호
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• pp.214-216
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• 2006

Isorhamnetin from the aerial parts of Persicaria thunbergii has been reported to have anti-tumor activity mediated by the inhibition of farnesyl protein transferase. In this study, we investigated the anti-proliferation effects of isorhamnetin on NIH3T3, K-RAS, H-RAS and SW620 cells, and it showed anti-proliferative effects in a dose-dependent manner with $IC_{50}$ value 4.1, 7.9, 20.2, and $22.4{\mu}g/ml$, repectively.

• Molecules and Cells
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• 제27권1호
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• pp.39-45
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• 2009

Ligand-dependent or independent oligomerization of receptor protein tyrosine kinase (RPTK) is often an essential step for receptor activation and intracellular signaling. The novel oncogene with kinase-domain (NOK) is a unique RPTK that almost completely lacks an ectodomain, expresses intracellularly and activates constitutively. However, it is unknown whether NOK can form oligomer or what function oligomerization would have. In this study, two NOK deletion mutants were generated by either removing the ectodomain ($NOK{\Delta}ECD$) or including the endodomain (NOK-ICD). Co-immunoprecipitation demonstrated that the transmembrane (TM) domain of NOK was essential for its intermolecular interaction. The results further showed that NOK aggregated more closely as lower order oligomers (the dimer- and trimer-sized) than either deletion mutant did since NOK could be crosslinked by both Sulfo-EGS and formaldehyde, whereas either deletion mutant was only sensitive to Sulfo-EGS. Removing the NOK TM domain (NOK-ICD) not only markedly promoted higher order oligomerization, but also altered the subcellular localization of NOK and dramatically elevated the NOK-mediated constitutive activation of extracellular signal-regulated kinase (ERK). Moreover, NOK-ICD but not NOK or $NOK{\Delta}ECD$ was co-localized with the upstream signaling molecule RAS on cell membrane. Thus, TM-mediated intermolecular contacting may be mainly responsible for the constitutive activation of NOK and contribute to the autoinhibitory effect on RAS/MAPK signaling.

• Asian Pacific Journal of Cancer Prevention
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• 제16권3호
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• pp.1077-1082
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• 2015

Recent studies have suggested that the RAS protein activator like-1 (RASAL1) functions as a tumor suppressor in vitro and may play an important role in the development of gastric cancer. However, whether or not RASAL1 suppresses tumor growth in vivo remains to be determined. In the present study, we investigated the role of RASAL1 in gastric carcinogenesis using an in vivo xenograft model. A lentiviral RASAL1 expression vector was constructed and utilized to transfect the human poorly differentiated gastric adenocarcinoma cell line, BGC-823. RASAL1 expression levels were verified by quantitative real-time RT-PCR and Western blotting analysis. Then, we established the nude mice xenograft model using BGC-823 cells either over-expressing RASAL1 or normal. After three weeks, the results showed that the over-expression of RASAL1 led to a significant reduction in both tumor volume and weight compared with the other two control groups. Furthermore, in xenograft tissues the increased expression of RASAL1 in BGC-823 cells caused decreased expression of p-ERK1/2, a downstream moleculein the RAS/RAF/MEK/ERK signal pathway. These findings demonstrated that the over-expression of RASAL1 could inhibit the growth of gastric cancer by inactivation of the RAS/RAF/MEK/ERK pathway in vivo. This study indicates that RASAL1 may attenuate gastric carcinogenesis.

• 한국양식학회지
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• 제21권3호
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• pp.125-132
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• 2008

Seed production of olive flounder Paralichthys olivaceus was performed in a pilot RAS. The growth of juvenile olive flounder and changes in water quality were monitored for the entire production period. The pilot RAS consisted of 8 circular culture tanks($4.0mD{\times}1.0mH$), 2 trickling biofilters($1.7mD{\times}2.0mH$), 2 protein skimmers ($0.8mD{\times}2.5mH$), and 4 sedimentation chambers($0.7mD{\times}1.5mH$). The culture surface area was about $100.5m^2$ and the actual working volume was about $106.9m^3$. As many as 300,000 fertilized olive flounder eggs were initially distributed into 2 culture tanks with the water temperature at $19.0^{\circ}C$. Live feeds such as rotifers and Artemia nauplii were fed until the 32nd day after hatching, and a commercial diet was fed from the 19th day to the end of the experiment. After 70 days, 150,256 juveniles with a body length of $65.8{\pm}3.9mm$ were produced in the RAS, with a daily growth rate for body length of 4.7%/day. At this time, the final culture density was 1,495 individuals $m^{-2}$, and 13.6 L of makeup water, 0.071 kW of electricity and 0.025 L of diesel fuel were used to produce a juvenile olive flounder. During metamorphosis of the larvae, the TAN concentration increased to 0.99 mg/L, which made the larvae sensitive to result in some mortality. However no more massive mortality occurred at the juvenile stage after metamorphosis even at a TAN concentration of 4.25 mg/L and a ${NO_2}^{-}-N$ concentration of 2.45 mg/L.

• Bulletin of the Korean Chemical Society
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• 제27권4호
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• pp.529-534
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• 2006

Chemical modification of the S. cerevisiae farnesyl protein transferase (FPT) with CMC, phenylglyoxal and DEPC resulted in enzyme inactivation, depending upon the reagent concentration. The peptide substrate GST-PEP-I, a GST-fused undecapeptide mimicking the C-terminus of $p21^{Ki-ras}$, protected the enzyme against inactivation by CMC which is specific to either aspartate or glutamate, while the other substrate farnesyl pyrophosphate (FPP) showed protection against phenylglyoxal which is the specific modifier of arginine residues, dependent on the substrate concentrations. Neither of the two substrates protected the enzyme against histidine inactivation by DEPC. It is suggested that there is at least one aspartate or glutamate residue at the peptide substrate binding site, and that at least one arginine residue is located at the binding site of FPP. There also seems to be at least one histidine residue which is critical for enzymic activity and is exposed toward the bulk solution, excluded from the substrate binding sites.

• BMB Reports
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• 제51권12호
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• pp.611-612
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• 2018

$C1q/TNF-{\alpha}-Related$ Protein 1 (CTRP1) has recently been shown to act as a blood pressure regulator, as it induces vasoconstriction. In the aorta, CTRP1 facilitates recruitment of angiotensin II receptor 1 (AT1R) to plasma membrane, through activation of the AKT/AS160 signaling pathway. This leads to activation of the Ras homolog gene family (Rho)/Rho kinase (ROCK) signaling pathway, resulting in vasoconstriction. Accordingly, mice overexpressing Ctrp1 have hypertensive phenotype. Patients with hypertension also display higher circulating CTRP1 levels, compared to healthy individuals, indicating that excessive CTRP1 may affect development of hypertension. Conversely, CTRP1 is regarded as an 'innate blood pressure modulator' because CTRP1 increases blood pressure under dehydration to prevent hypotension. Mice lacking Ctrp1 fail to maintain normotension under dehydration conditions, resulting in hypotension, suggesting that CTRP1 is an essential protein for maintaining blood pressure homeostasis. In conclusion, CTRP1 is a novel, anti-hypotensive vasoconstrictor that increases blood pressure during dehydration-induced hypotension.

• Fisheries and Aquatic Sciences
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• 제21권2호
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• pp.4.1-4.5
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• 2018

Insulin-like growth factors (IGFs), along with IGF-binding protein and IGF receptor, are well-known regulators in the growth and survival of vertebrates. In this study, we investigated the involvement of IGFs and protein variation during embryonic development of the olive flounder (Paralichthys olivaceus). Morphological stages were divided into six main developments as blastula, gastrula, cephalization, cranial regionalization, tail lift, and hatch. During embryonic development, protein variation was investigated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrospray ionization quadrupole time-of-flight mass spectrometry/mass spectrometry. In addition, the mechanism of signaling of IGF-I receptor was examined using immuno-blot analysis. We found marked changes in protein expression at four stages of embryonic development and identified proteins as belonging to the vitellogenin 2 family. As development progresses, expression of IGF-II, phosphotyrosine, and phospho-Akt increased, while expression of growth factor receptor-bound protein 2 (GRB2) and one of guanine-nucleotide-binding proteins (Ras) decreased. These results provide basic information on the IGF system in the embryonic development of the olive flounder.

• 생명과학회지
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• 제26권12호
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• pp.1383-1391
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• 2016

타목시펜과 같은 항에스트로젠은 ER 양성의 초기 유방암 환자에게 사용되고 있다. 그러나 대부분의 환자에서 이 항에스트로젠에 대한 내성 발현은 불가피하게 발생한다. BCAR3 유전자는 사람의 에스트로젠 의존성 유방암에서 tamoxifen 내성유도를 야기하는 단백질로 발견되었다. 우리들은 이전에 이 BCAR3 유전자가 세포주기 진행과 EGF와 인슐린에 의한 DNA 합성 신호전달경로를 조절한다고 보고하였다. 본 연구에서는, 비종양성 정상적인 인간유방상피세포인 MCF-12A세포에서 c-Jun 전자의 조절에 대한 BCAR3유전자의 기능적인 역할을 조사하였다. BCAR3의 일시적인 발현 또는 지속적인 발현이 c-Jun mRNA와 단백질의 발현을 증가하는 것을 발견하였다. 또한 BCAR3 발현 유전자의 미세주사에 의해 세포 증식이 증가하였다. 이 c-Jun의 발현 증가는 promoter의 활성화를 통해 일어난다. 또한 BCAR3에 의한 c-Jun 발현 유도가 억제성 Ras, Rac, Rho에 의해 억제되었다. 다음으로 EGF 성장인자에 의한 c-Jun 발현 유도에 대한 BCAR3의 영향을 단일 세포 미세주사법에 의해 조사하였다. BCAR3 항체, BCAR3의 siRNA와 같은 BCAR3의 기능을 억제할 수 있는 물질들을 세포로 미세주사하면 EGF에 의한 c-Jun의 발현을 억제하였지만, IGF-1 성장인자에 의한 c-Jun 발현은 억제하지 않았다. 이러한 결과들로부터 BCAR3는 c-Jun 단백질 발현 유도와 세포 증식에 중요한 역할을 하며, 여기에는 Ras, Rac, Rho와 같은 GTPase들이 필요하다는 것을 발견하였다.

• 순환기질환의공학회:학술대회논문집
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• 순환기질환의공학회 2006년도 제6회 학술대회
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• pp.10-13
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• 2006

Inflammation plays an important role in the progression of atherosclerosis and plaque destabilization converting a chronic process into an acute disorder with ensuing thromboembolism. Current therapeutic effective in preventing atherosclerosis and stroke such as statins, ASS and RAS inhibitors my exert part of their effects by modulating inflammatory responses in the vessel walls. As alternative approaches, discovery to find having inhibitory action of MMP activity, COX-2, macrophage infiltration, such as APE1/ref-1 and fusion technology for cell permeable protein may provide a new antiatherosclerotic therapy in the future.