• BMB Reports
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• 제45권8호
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• pp.470-475
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• 2012

Protein arginine methyltransferase 1 (PRMT1), a type-I arginine methyltransferase, has been implicated in diverse cellular events. We have focused on the role of PRMT1 in gliomagenesis. In this study, we showed that PRMT1 expression was up-regulated in glioma tissues and cell lines compared with normal brain tissues. The knock-down of PRMT1 resulted in an arrest in the G1-S phase of the cell cycle, proliferation inhibition and apoptosis induction in four glioma cell lines (T98G, U87MG, U251, and A172). Moreover, an in vivo study confirmed that the tumor growth in nude mouse xenografts was significantly decreased in the RNAi-PRMT1 group. Additionally, we found that the level of the asymmetric dimethylated modification of H4R3, a substrate of PRMT1, was higher in glioma cells than in normal brain tissues and decreased after PRMT1 knock-down. Our data suggest a potential role for PRMT1 as a novel biomarker of and therapeutic target in gliomas.

• Asian Pacific Journal of Cancer Prevention
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• 제13권11호
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• pp.5865-5869
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• 2012

RHOXF1 has been shown to be expressed in embryonic stem cells, adult germline stem cells and some cancer lines. It has been proposed as a candidate gene to encode transcription factors regulating downstream genes in the human testis with antiapoptotic effects. Its expression in cancer cell lines has implied a similar role in the process of tumorigenesis. The human breast cancer cell lines MDA-MB-231 and MCF-7 were cultured in DMEM medium and transfected with a pGFP-V-RS plasmid bearing an RHOXF1 specific shRNA. Quantitative real-time RT-PCR was performed for RHOXF1, CASP8, BCL2 and HPRT genes. Decreased RHOXF1 expression was confirmed in cells after transfection. shRNA knock down of RHOXF1 resulted in significantly decreased BCL2 expression in both cell lines but no change in CASP8 expression. shRNA targeting RHOXF1 was shown to specifically mediate RHOXF1 gene silencing, so RHOXF1 can mediate transcriptional activation of the BCL2 in cancers and may render tumor cells resistant to apoptotic cell death induced by anticancer therapy. shRNA mediated knock down of RHOXF1 can be effective in induction of apoptotic pathway in cancer cells via BCL2 downregulation, so it can have potential therapeutic utility for human breast cancer.

• The Korean Journal of Physiology and Pharmacology
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• 제18권6호
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• pp.525-530
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• 2014

Transient receptor potential vanilloid subtype 1 (TRPV1) was originally found in sensory neurons. Recently, it has been reported that TRPV1 is expressed in salivary gland epithelial cells (SGEC). However, the physiological role of TRPV1 in salivary secretion remains to be elucidated. We found that TRPV1 is expressed in mouse and human submandibular glands (SMG) and HSG cells, originated from human submandibular gland ducts at both mRNA and protein levels. However, capsaicin (CAP), TRPV1 agonist, had little effect on intracellular free calcium concentration ($[Ca^{2+}]_i$) in these cells, although carbachol consistently increased $[Ca^{2+}]_i$. Exposure of cells to high temperature (> $43^{\circ}C$) or acidic bath solution (pH5.4) did not increase $[Ca^{2+}]_i$, either. We further examined the role of TRPV1 in salivary secretion using TRPV1 knock-out mice. There was no significant difference in the pilocarpine (PILO)-induced salivary flow rate between wild-type and TRPV1 knock-out mice. Saliva flow rate also showed insignificant change in the mice treated with PILO plus CAP compared with that in mice treated with PILO alone. Taken together, our results suggest that although TRPV1 is expressed in SGEC, it appears not to play any direct roles in saliva secretion via transcellular pathway.

• 통합자연과학논문집
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• 제12권4호
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• pp.142-146
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• 2019

Peroxidasin is a unique member of peroxidase family in that it has extracellular matrix (ECM) motif as well as peroxidase activity. Peroxidasins are involved in consolidation the extracellular matrix during development and in innate immune defense. C. elegans has two functional peroxidasins, PXN-1 and PXN-2, and PXN-2 is known to contribute to innate immune system. However, it is not clear of PXN-1 function in innate immune system. Therefore, this study is focused on the function of PXN-1 and the relationship between PXN-1 and PXN-2 in innate defense system in C. elegans. When pxn-1 was knocked down by RNAi, the worm turned to be more resistant to pathogens, Staphylococcus aureus and Pseudomonas aeruginosa and the enhanced resistance was abolished in pxn-1pxn-2 double knock down. By contrast, pxn-2 knock down showed stronger susceptibility to the pathogens. These results suggest that pxn-2 can contribute the pathogen resistance and pxn-1 can suppress the pathogen resistance. To confirm the idea, overexpression experiments were performed. Overexpression of pxn-1 showed more susceptible to pathogens compared to the control and double overexpression of pxn-1pxn-2 overcame the susceptibility of pxn-1 overexpression to the pathogens. On the other hand, pxn-2 overexpression made the worm more resistant to the pathogens and the resistance was maintained in pxn-1pxn-2 double overexpression. The comparison of the susceptibilities to the bacterial pathogens in above mentioned constructs suggests that PXN-1 suppress the function of PXN-2 in defense against bacterial pathogens in Caenorhabditis elegans.

• Asian Pacific Journal of Cancer Prevention
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• 제14권11호
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• pp.6625-6629
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• 2013

Breast cancer accounts for one third of new cancer cases among women. The need for biomarkers for early detection is the stimulus to researchers to evaluate altered expression of genes in tumours. Cancer-testis (CT) genes are a group with limited expression in normal tissues except testis but up-regulation in a wide variety of cancers. We here evaluated expression of two CT genes named FBXO39 and TDRD4 in 32 invasive ductal carcinoma samples, 10 fibroadenomas and 6 normal breast tissue samples, in addition to two breast cancer cell lines, MCF-7 and MDA-MB-231, by the means of quantitative real time RT-PCR. FBXO39 showed significant up-regulation in invasive ductal carcinoma samples in comparison with normal samples. It also was expressed in both cell lines and after RHOXF1 gene knock down it was down-regulated in MCF-7 but up-regulated in the MDA-MB-231 cell line. TDRD4 was not expressed in the MCF-7 cell line and any of the tissue samples except testis. However, it was expressed in MDA-MB-231 and was up-regulated after RHOXF1 gene knock down. Our results show that FBXO39 but not TDRD4 can be used for cancer detection and if proved to be immunogenic, might be a putative candidate for breast cancer immunotherapy.

• Journal of Korean Biological Nursing Science
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• 제22권1호
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• pp.45-52
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• 2020

Purpose: The purpose of this study was to investigate the risk factors of intracranial hemorrhage in children with skull fractures from head trauma. Methods: The retrospective study included 205 patients diagnosed with a skull fracture in a pediatric emergency room. Data were analyzed using 𝓍²-test, Fisher's exact test, t-test, and logistic regression analysis with the SPSS/WIN24.0 program. Results: Intracranial hemorrhage was diagnosed in 71 patients. There were statistically significant differences between the hemorrhagic group and non-hemorrhagic group in age group, places of accident, type of accident, location of the fracture, and symptoms. Intracranial hemorrhage by age group was higher in school-age and adolescence than in infancy. The places of accidents of hemorrhage were higher in street and school than in the home. The types of an accident of bleeding were higher in the case of knock and traffic accident than in fall. Symptoms of nausea, headache, and loss of consciousness were associated with higher intracranial hemorrhage. Multivariable logistic regression analysis showed that knock (OR= 3.29, 95% CI= 1.50-7.22), traffic accident (OR= 4.78, 95% CI= 1.31-17.43), nausea (OR= 4.18, 95% CI= 1.42-12.31), and loss of consciousness (OR= 3.29, 95% CI= 1.41-9.50) were risk factors for intracranial hemorrhage. Conclusion: In this study, the risk factors of intracranial hemorrhage were identified in pediatric patients with skull fractures caused by head trauma. It is recommended that the results of this study be used to manage and educate patients, caregivers, and medical staff after head trauma hemorrhage.

• 한국동물생명공학회지
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• 제36권3호
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• pp.121-128
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• 2021

Increasing the efficiency of HR (homologous recombination) is important for a successful knock-in. Rad51 is mainly involved in homologous recombination and is associated with strand invasion. The HR-related mismatch repair system maintains HR fidelity by heteroduplex rejection and repair. Therefore, the purpose of this study is to control Rad51, which plays a critical role in HR, through UV-induced DNA damage. It is also to confirm the effect on the expression of MMR related genes (Msh2, Msh3, Msh6, Mlh1, Pms2) and HR-related genes closely related to HR through treatment with the MMR inhibitor CdCl₂. The mRNA expression of Rad51 gene was confirmed in both HC11 cells and mouse testes, but the mRNA expression of Dmc1 gene was confirmed only in mouse testes. The protein expression of Rad51 and Dmc1 gene increased in UV-irradiated HC11 cells. After 72 hours of treatment with 1 ㎛ of CdCl₂, the mRNA expression level of Msh3, Pms2, and Rad51 decreased, but the mRNA expression level of Msh6 and Mlh1 increased in HC11 cells. There was no significant difference in Msh2 mRNA expression between CdCl₂ untreated-group and the 72 hours treated group. In conclusion, HR-related gene (Rad51) was increased by UV-induced DNA damage. Treatment of the MMR inhibitor CdCl₂ in HC11 cells decreased the mRNA expression of Rad51.

• 대한임상검사과학회지
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• 제48권2호
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• pp.114-117
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• 2016

골은 일생에 걸쳐 지속적인 재형성과장을 거치면서 유지되고 이러한 기전에 대한 연구는 골다공증을 비롯한 골대사 질환의 병태생리와 치료에 있어 큰 진전을 이루고 있다. 특히 생체 내 골형성 및 재생과정을 연구하는데 있어 형광표지자를 이용하는 방법이 널리 알려져 있는데, 그 중 calcein은 칼슘 킬레이터로 골이 새롭게 형성하는 부위에 녹색을 띔으로써 유용한 마커로 사용된다. 그러나 대부분의 골형성 연구에서 실험동물의 경우 표본제작을 할 때 크기가 작고 뼈가 부숴지기 쉬워 rat이나 rabbit을 이용하였으며, mice의 femur를 cross-section해서 관찰한 연구는 거의 없는 실정이다. 그래서 본 연구에서는 어린 mice를 실험동물로 이용하였으며, 생체 내 calcein을 4주간, 8주간 투여한 후 골 형성 변화를 형광현미경으로 관찰한 결과 8주차 쥐에서 4주차보다 진하고 골 형성 간격도 넓게 관찰된 것을 확인 할 수 있었다. Mice는 빠른 시일 내에 결과를 얻을 수 있고 부작용이 적은 장점이 있어서 앞으로 knock-out mice를 이용한 생체 내 실험에 활용하기 적합하다고 생각되며, 골형성 속도 평가 등 다양한 분야에서의 골 형성과 재생연구에 있어 기초 정보를 제공할 것으로 기대한다.

• Molecules and Cells
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• 제28권5호
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• pp.463-472
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• 2009

Although the possible cellular roles of several ubiquitin-specific proteases (UBPs) were identified in Arabidopsis, almost nothing is known about UBP homologs in rice, a monocot model plant. In this report, we searched the rice genome database (http://signal.salk.edu/cgi-bin/RiceGE) and identified 21 putative UBP family members (OsUBPs) in the rice genome. These OsUBP genes each contain a ubiquitin carboxyl-terminal hydrolase (UCH) domain with highly conserved Cys and His boxes and were subdivided into 9 groups based on their sequence identities and domain structures. RT-PCR analysis indicated that rice OsUBP genes are expressed at varying degrees in different rice tissues. We isolated a full-length cDNA clone for OsUBP6, which possesses not only a UCH domain, but also an N-terminal ubiquitin motif. Bacterially expressed OsUBP6 was capable of dismantling K48-linked tetra-ubiquitin chains in vitro. Quantitative real-time RT-PCR indicated that OsUBP6 is constitutively expressed in different tissues of rice plants. An in vivo targeting experiment showed that OsUBP6 is predominantly localized to the nucleus in onion epidermal cells. We also examined how knock-out of OsUBP6 affects developmental growth of rice plants. Although homozygous T3 osubp6 T-DNA insertion mutant seedlings displayed slower growth relative to wild type seedlings, mature mutant plants appeared to be normal. These results raise the possibility that loss of OsUBP6 is functionally compensated for by an as-yet unknown OsUBP homolog during later stages of development in rice plants.

• Asian-Australasian Journal of Animal Sciences
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• 제23권6호
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• pp.806-813
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• 2010

Animal bioreactors have been regarded as alternative tools for the production of limited human therapeutic proteins. The mammary glands of cattle are optimal tissues to produce therapeutic proteins that cannot be produced in large amounts in traditional systems based on microorganisms and eukaryotic cells. In this study, two knock-in vectors, pBCTPOKI-6 and pBCTPOKI-10, which target the hTPO gene on the bovine beta-casein locus, were designed to develop cloned transgenic cattle. The pBCTPOKI-6 and pBCTPOKI-10 vectors expressed hTPO protein in culture medium at a concentration of 774 pg/ml and 1,867 pg/ml, respectively. Successfully, two targeted cell clones were obtained from the bovine fibroblasts transfected with the pBCTPOKI-6 vector. Cloned embryos reconstructed with the targeted nuclei showed a lower in vitro developmental competence than those with the wild-type nuclei. After transfer of the cloned embryos into recipients, 7 pregnancies were detected at 40 to 60 days of gestation, but failed to develop to term. The results are the first trial for targeting of a human gene on the bovine milk protein gene locus, providing the potential for a large-scale production of therapeutic proteins in the animal bioreactor system.