• Asian-Australasian Journal of Animal Sciences
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• 제15권7호
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• pp.1066-1070
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• 2002

10 male piglets at 5-6 weeks old with similar body weight (BW) were randomly assigned into the experimental (EXP) and control (CON) groups. The animals in EXP received intro-muscular injection with daidzein (DA) at the dose of 0.5 mg DA per kg start BW on day 1. The same procedures were repeated once every 3 days for eight times. The animals in CON received the injection only with same volume of control peanut oil. The animals were weighted on day 14 and 28 and the blood samples were obtained at different stages of the treatment for determining IGF-I levels and blood parameters. At the end of the experiment, the thymus and spleen from all the animals were surgically taken out and weighted. The results showed that BW and average daily gain (ADG) were not significantly different between the groups in term of the whole period, but ADG between days 14-28 was higher in EXP than in CON (p<0.05). On days 18, 21 and 25, IGF-I levels in EXP group were 20.53% (p<0.05), 15.92% (p>0.05) and 23.46% (p<0.05), respectively, higher than those in CON. The weights of thymus and spleen, the ratios of their weights to BW and red blood count (RBC) did not significantly differ between the groups at all stages. White blood count (WBC) in EXP steadily increased from day 22, reached its apex on day 24, which was higher than in CON (p<0.05) and its own levels on day 20 and 22 (p<0.01 or p<0.05), and remained higher on the later time (p=0.058). The results of percentage of T-Lymphocytes also demonstrated similar trend to WBC, but TLymphocyte transformation rate (%) appeared no significant change between the groups. In conclusion, Daidzein could stimulate male piglet growth and elevate serum IGF-I levels at certain stages of the treatment. It could also increase WBC and T-Lymphocyte rates, but had no significant impacts on RBC and T-Lymphocyte transformation rate.

• 치과방사선
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• 제28권1호
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• pp.245-259
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• 1998

Cellular transforming genes have been identified in a number of different tumor cell lines and tumor types. A significant number of these oncogenes belong to the ras gene family. The ras gene family consists of three closely related genes:H-ras, K-ras and N-ras which code for a related 21 kDa protein. Mutations in codon 12, 13 and 61 of one of the three ras genes convert these genes into acute oncogenes. The presence of H-ras gene mutations has important prognostic implications in various tumors. Each genomic DNA was isolated from tumors induced by implantation with DMBA, or by treatment with DMBA -implantation/irradiation. When genome DNA was transfected into NIH 3T3 cells and investigated by two-step PCR-RFLP, the fOllowing results were concluded: 1. Transformation foci developed in two groups when the genome DNA of two experimental groups were transfected into NIH 3T3 cells. 2. Transformation efficiency was 0.01-0.02 foci/㎍DNA in the experimental group with the DMBA-implantation, 0.01-0.03 foci/㎍lgDNA in the experimental group with the DMBA-implantation/irradiation according to results of transfection assay. 3. When the point mutation of H-ras gene was investigated by a two-step PCR-RFLP, there was 13.9％ (5/36) in the experimental group with the DMBA implantation, 15.4 ％ (6/39) in the experimental group with the DMBA -implantation/irradiation. 4. The point mutation in codon 12 and 61 of H-ras was 5.6％(2/36) and 8.3％(3/36) in the experimental group with the DMBA implantation. 5. The point mutation in codon 12 and 61 of H-ras gene was 7.7％(3/39) in the experimental group with the DMBA -implantation/irradiation.

• 자원환경지질
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• 제52권6호
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• pp.627-635
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• 2019

슬레이트는 석면을 이용한 대표적인 건축자재 중 하나로써 백석면(10~20%)과 시멘트 성분을 결합하여 만든 제품이다. 슬레이트에 포함되어 있는 석면은 인체에 유입되면 세포 손상이나 변형을 일으키고 체외로 잘 배출되지 않아 폐암, 석면폐, 악성중피종 및 흉막비후 등과 같은 질병을 일으키는 원인이 되는 것으로 입증되어 1977년 세계보건기구(WHO) 산하 국제암연구소(IARC)에서는 1군 발암물질로 지정하였다. 현재 이러한 슬레이트는 대부분 지정매립장에 매립하여 처리하고 있으나 매립용량이 한계에 다다르고 있고 매립한다고 하여도 추후 외부환경으로 노출될 수 있는 잠재적인 위험성이 있어 매립 처리방법 이외에 슬레이트에 포함된 석면을 무해화하여 안전하게 처리할 수 있는 방법이 필요하다. 따라서 이 연구에서는 발열반응 촉매제와 열처리를 이용하여 슬레이트에 함유된 석면 무해화 가능성을 확인하고자 하였다. 실험은 석면해체·제거 사업장에서 발생한 석면함유 슬레이트를 이용하였고 발열반응 촉매제는 염화칼슘(CaCl₂), 염화마그네슘(MgCl₂), 수산화나트륨(NaOH), 규산소듐(Na₂SiO₃), 카올린[Al₂Si₂O₅(OH)₄)], 활석[Mg₃Si₄O₁₀(OH)₂]을 이용하여 총 6가지의 촉매제를 제조하였다. 6가지의 촉매제를 슬레이트에 각각 도포한 후 열중량-시차열분석(TG-DTA)을 실시하여 분석결과를 토대로 슬레이트 무해화를 위한 열처리 온도를 750℃로 결정하였다. 슬레이트에 6가지 촉매제를 각각 도포한 후 750℃에서 2시간 열처리하여 X-선 회절 분석(XRD), 주사전자현미경 분석(SEM-EDS), 투과전자현미경 분석(TEM-EDS)을 한 결과 슬레이트 내 백석면[chrysotile, Mg₃Si₂O₅(OH₅)]이 주상의 고토감람석(forsterite, Mg₂SiO₄)으로 상전이 됨을 확인하였다. 또한, 슬레이트 원시료와 발열반응 촉매제 도포 후 열처리한 시료에 물리적인 힘을 가하여 광물의 형상 변화를 비교 관찰한 결과, 슬레이트 내 백석면은 섬유형을 유지하였으나 촉매제 도포 및 열처리를 한 시료는 무정형 형태로 깨지는 것을 확인할 수 있었다. 따라서 발열반응 촉매제와 열처리를 통하여 낮은 온도에서 경제적으로 석면함유 슬레이트를 안전하게 처리할 수 있는 하나의 방안을 제시할 수 있을 것으로 사료된다.

• Animal cells and systems
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• 제1권3호
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• pp.521-526
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• 1997

We isolated a mouse nck cDNA from the thymus cDNA expression library. The cDNA encodes a 377 amino acid protein and displays 97% amino acid sequence identity to human oncogenic protein nck, which is composed almost exclusivelv of three src homology 3 (SH3) domains and one SH2 domain. The sequence analysis also showed that the isolated cDNA is the mouse counterpart of the human nck and different from the mouse grb4, which has been reported to be highly similar to the human nck and, therefore considered as a mouse nck, Northern blot analysis showed that the transcript of the gene was 1.8 kb and was highly expressed in the testis, thymus, and brain but moderately in the liver and lymph node. Western blot analysis showed that the size of the protein was about 47 kDa. Overexpression of the mouse Nck transformed a mouse fibroblast cell line, NIH3T3. The results clearly indicate that normal nck gene has transforming ability and provide an argument against a suggested possibility that the transforming ability of the human nck gene is due to a mutation(s) in the gene.

• Toxicological Research
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• 제14권2호
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• pp.123-127
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• 1998

BRCA1 is a tumor suppresser gene in familial cases of breast cancer. It has been controversial whether the subcellular localization of BRCA1 is located in nuclei or cytoplasm in normal human breast cells. We found that a p220 protein was expressed in Type II Normal human breast epithelial cells (NHBEC) but not in Type I NHBEC in Western blot analysis using the 17F8 (3A2) antibody. Immunostaining using the same antibody revealed positive staining in nuclei, cytoplasm and perinuclei of Type II cells and negative staining in Type I NHBEC. The p220 protein, however, was expressed in SV40 immortalized Type I NHBEC and tumorigenic cells derived from them after x-ray and neu oncogene treatment. The subcelluar localization was mostly cytoplasmic and punctate in the nuclei. The breast carcinoma cell lines, MCF-7 and T47D, also expressed the p220 protein. Using RT-PCR, we observed the expression of BRCA1 mRNA in both Type I and Type II NHBEC. This result indicated that there might be mechanisms involved in post-translational or translational regulation of BRCA1 gene. It is speculated that the absence of BRCA1 protein expression in Type I NHBEC might playa role in their susceptibility to neoplastic transformation.

• Biomolecules & Therapeutics
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• 제17권4호
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• pp.438-445
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• 2009

We investigated global gene expression from both mouse liver and mouse hepatic cell lines treated with acetaminophen (APAP) in order to compare in vivo and in vitro profiles and to assess the feasibility of the two systems. During our analyses of gene expression profiles, we picked up several down-regulated genes, such as the cytochrome P450 family 51 (Cyp51), sulfotransferase family cytosolic 1C member 2 (Sult1c2), 3-hydroxy-3-methylglutaryl-Coenzyme A synthase 1 (Hmgcs1), and several genes that were up-regulated by APAP, such as growth arrest and DNA-damage-inducible 45 alpha (Gadd45a), transformation related protein 53 inducible nuclear protein 1 (Trp53inp1) and zinc finger protein 688 (Zfp688). For validation of gene function, synthesized short interfering RNAs (siRNAs) for these genes were transfected in a mouse hepatic cell line, BNL CL.2, for investigation of cell viability and mRNA expression level. We found that siRNA transfection of these genes induced down-regulation of respective mRNA expression and decreased cell viability. siRNA transfection for Cyp51 and others induced morphological alterations, such as membrane thickening and nuclear condensation. Taken together, siRNA transfection of these six genes decreased cell viability and induced alteration in cellular morphology, along with effective inhibition of respective mRNA, suggesting that these genes could be associated with APAP-induced toxicity. Furthermore, these genes may be used in the investigation of hepatotoxicity, for better understanding of its mechanism.

• Journal of Periodontal and Implant Science
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• 제40권1호
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• pp.39-44
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• 2010

Purpose: Bone tissues for clinical application can be improved by studies on osteoblast differentiation. Runx2 is known to be an important transcription factor for osteoblast differentiation. However, bone morphogenetic protein (BMP)-2 treatment to stimulate Runx2 is not sufficient to acquire enough bone formation in osteoblasts. Therefore, it is necessary to find other regulatory factors which can improve the transcriptional activity of Runx2. The erythroblast transformation-specific (ETS) transcription factor family is reported to be involved in various aspects of cellular proliferation and differentiation. Methods: We have noticed that the promoters of osteoblast differentiation markers such as alkaline phosphatase (Alp), osteopontin (Opn), and osteocalcin (Oc) contain Ets binding sequences which are also close to Runx2 binding elements. Luciferase assays were performed to measure the promoter activities of these osteoblast differentiation markers after the transfection of Runx2, myeloid Elf-1-like factor (MEF), and Runxs+MEF. Reverse-transcription polymerase chain reaction was also done to check the mRNA levels of Opn after Runx2 and MEF transfection into rat osteoblast (ROS) cells. Results: We have found that MEF, an Ets transcription factor, increased the transcriptional activities of Alp, Opn, and Oc. The addition of Runx2 resulted in the 2- to 6-fold increase of the activities. This means that these two transcription factors have a synergistic effect on the osteoblast differentiation markers. Furthermore, early introduction of these two Runx2 and MEF factors significantly elevated the expression of the Opn mRNA levels in ROS cells. We also showed that Runx2 and MEF proteins physically interact with each other. Conclusions: Runx2 interacts with MEF proteins and binds to the promoters of the osteoblast markers such as Opn nearby MEF to increase its transcriptional activity. Our results also imply that osteoblast differentiation and bone formation can be increased by activating MEF to elicit the synergistic effect of Runx2 and MEF.

• 한국환경성돌연변이발암원학회지
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• 제21권2호
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• pp.164-168
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• 2001

Abel et al. in Germany discovered a new dioxin-responsive gene, which has later been identified as rpt-1 (regulatory protein T-lymphocyte 1). While it is speculated that rpt-1 may play a role in signal transduction and carcinogenesis, its roles and functions remain unknown. The present study attempted to analyze functions of rpt-1 in human epithelial cells following the xenobiotic exposures. While German counterpart analyzed expressionn of rpt-1 in spleen and thymus cells from mouse and rat and characterizes molecular properties of the gene, our work mainly focused on analyzing function of rpt-1 in human skin cells. Expression of rpt-1 in human cells were analyzed by western and northern blot RT-PCR analysis. Expression of rpt-1 as well as Staf-50 in human cells with or without exposure to environmental pollutants were also analyzed by northern blot analysis, since Staf-50 is homologous with rpt-1 and found in human cells. To help study roles of rpt-1 in human cell system, retroviral vector system carrying rpt-1 gene under the CMV promoter were constructed and transfected. Cells overexpressing the gene after the transfection showed an increase of cell density and soft agar colony formations, as compared to the control cells, suggesting that rpt-1 may play a certain role in the transformation processes of human cells. While the expression of rpt-1 in spleen and thymus is known to be strong in the laboratory animals, both the basal and TCDD-induced expression of rpt-1 in the current cellular system remained insignificant. It is speculated that the expression pattern of rpt-1 may be tissue- and species-specific. The present study demonstrated a strong expression of rpt-1 protein in the brain of SD rat model. Since there is no previous report on the expression of rpt-1 in the brain tissue, the result may play a significant role in understanding dioxin-induced neurotoxicities in the future. The present study provides an opportunity to understand a role of rpt-1 in human cell system and suggest a possible lead and basis for the future study of dioxin-induced neurotoxicities.

• 한국수정란이식학회지
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• 제26권4호
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• pp.229-235
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• 2011

Sloan-Kettering virus gene product of a cellular protooncogene c-Ski is an unique nuclear pro-oncoprotein and belongs to the Ski/Sno proto-oncogene family. Ski plays multiple roles in a variety of cell types, it can induce both oncogenic transformation and terminal muscle differentiation when expressed at high levels. Ski protein is implicated in proliferation/differentiation in a variety of cells. The alternative fate of granulosa cells other than apoptosis is to differentiate to luteal cells, however, it is unknown whether Ski is expressed and has a role in granulosa cells undergoing luteinization. Thus, the aim of this study was, by means of immunohistochemical methods, to locate Ski protein in the rat ovaries during ovulation and corpora lutea (CL) formation to predict the possible involvement of Ski in luteinization. In addition, we performed to examine whether the initiation of luteinization with luteinizing hormone (LH) directly regulates expression of Ski in the luteinized granulosa and luteal cells after ovulation by in vivo models. In order to examine the expression pattern of Ski protein along with the progress of luteinization, follicular growth was induced by administration of equine chorionic gonadtropin to immature female rat, and luteinization was induced by human chorionic gonadtropin treatment to mimic luteinizing hormone (LH) surge. While no Ski-positive granulosa cells were present in preovulatory follicle, Ski protein expression was induced in response to LH surge, and was maintained after the formation of corpus luteum (CL). These results indicate that Ski is profoundly expressed in the luteinized granulosa cells and luteal cells of CL during luteinization, and suggest that Ski may play a role in luteinization of granulosa cells.

• 동의생리병리학회지
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• 제18권6호
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• pp.1613-1616
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• 2004

The cellular response to hypoxia is controlled to a large degree by the transcription factor Hypoxia-inducible factor-1(HIF-1). HIF-1 is a transcription factor that is activated by hypoxia and plays a critical role in the development of the cancer phenotype. HIF-1 regulates transcription of a number of genes crucial for tumor survival under hypoxic conditions, including vascular endothelial growth factor(VEGF), erythropoietin(Epo) and several glycolytic enzymes. Tumors in which hypoxia can not induce HIF-1 transcriptional activity remain small and fail to metastasize. In this study, we examined whether aqueous root extract of Ailanthus altissima (REA) downregulate HIF-1, VEGF and p53, and raise the possibility that depletion of these proteins and the anti proliferative activities of REA have any effects on inhibition of angiogenesis in MCF-7 cells. Pharmacologic targeting of specific signal transduction pathways related to oncogenic transformation is a promising approach in cancer treatment. Therefore, REA could be a candidate drug for further clinical development.