• Title/Summary/Keyword: Methods of Expression

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CD30-Mediated Regulation of Cell Adhesion Molecule Expression on Murine T Cells

  • Nam, Sang-Yun
    • IMMUNE NETWORK
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    • v.3 no.1
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    • pp.8-15
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    • 2003
  • Background: CD30 is a member of TNF receptor family and expressed on lymphocytes and other hematopoietic cells following activation as well as Hodgkin and Reed-Sternberg cells in Hodgkin's lymphoma. In this study, CD30-mediated regulation of cell adhesion molecule expression on normal activated mouse T cells was investigated. Methods: Mouse T cells were activated with anti-CD3 antibody for induction of CD30, which was cross-linked by immobilized anti-CD30 antibody. Results: High level of CD30 expression on T cells was observed on day 5, but only little on day 3 even under culture condition resulting in an identical T cell proliferation, indicating that CD30 expression requires a prolonged stimulation up to 5 days. Cross-linking of CD30 alone altered neither proliferation nor apoptosis of normal activated T cells. Instead, CD30 appeared to promote cell adherence to culture substrate, and considerably upregulated ICAM-1 and, to a lesser extent, ICAM-2 expression on activated T cells, whereas CD2 and CD18 (LFA-1) expression was not affected. None of cytokines known as main regulators of ICAM-1 expression on tissue cells (IL 4, $IFN{\gamma}$ and $IFN{\alpha}$) enhanced ICAM-1 expression in the absence of CD30 signals. On the other hand, addition of $NF-{\kappa}B$ inhibitor, PDTC (0.1 mM) completely abrogated the CD30-mediated upregulation of ICAM-1 expression, but not CD2 and ICAM-2 expression. Conclusion: This results support that CD30 upregulates ICAM-1 expression of T cell and such regulation is not mediated by higher cytokine production but $NF-{\kappa}B$ activation. Therefore, CD30 may play important roles in T-T or T-B cell interaction through regulation of ICAM-1, and -2 expression.

IFN-γ Regulates Expression of BRG1 Associated Factor 155/170 and Sensitivity to Steroid in Astrocytes

  • Lim, Jung-Hee;Lee, Jeonggi;Park, Joo Young;Choi, In-Hong
    • IMMUNE NETWORK
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    • v.4 no.4
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    • pp.224-228
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    • 2004
  • Background: The expression of BRG1 associated factors (BAF) 155 and BAF 170 in response to $IFN-{\gamma}$ or $TNF-{\alpha}$ was studied in astrocytoma cell lines and primary astrocytes. BAFs are complexed with BRG1 and are also associated with activated glucocorticoid for glucocorticoid trans-activation. Methods: $IFN-{\gamma}$ was pretreated for 18 hrs and cells were incubated with IL-1 or $TNF-{\alpha}$ for 72 hrs or 96 hrs with different concentrations of steroid. Cell death was measured by LDH assay. BAF expression was assayed by RT-PCR. Results: $IFN-{\gamma}$ increased cell death by dexamethasone in LN215 cells but not in LN319 cells. The $IFN-{\gamma}$ increased the expression of BAF 155 and BAF 170 in adult astrocytes and LN215 cells, but $IFN-{\gamma}$ decreased the expression of BAF 155/170 in LN319 cells. The effect of $IFN-{\gamma}$ on the expression of BAF was not as clear in fetal astrocytes as it was in adult astrocytes. Conclusion: Our results suggest cytokines produced during immune reaction or immunotherapy may modulate steroid susceptibility of astrocytes and astrocytoma cells by influencing the expression of BAFs.

Construction and Expression of an Eukaryotic Expression Vector Containing the IER3 Gene

  • Wang, Zhen;Yu, Hong-Sheng;Yao, Ru-Yong;Qiu, Wen-Sheng;Yue, Lu;Sui, Ai-Hua;Liu, Xiang-Ping;Liu, Shi-Hai
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.1
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    • pp.507-510
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    • 2013
  • Background: More and more research indicate that the immediately early response gene 3 (IER3) is involved inmany biological provesses, such as apoptosis and immunoreaction, as well as viral infection, tumorigenesis and tumour progression. Methods: Here we describe the construction of an eukaryotic expression vector containing IER3 gene and its expression in A549 cells as assessed through fluorescence microscopyand Western-blotting. Results: Fluorescence detection displayed that GFP in cytoplasm was high during 48 and 72 hours post-transfection. In addition, Western blotting showed significant increase in IER3 gene expression in the transfected cells compared with controls. Conclusion: The recombinate plasmid expression vector was constructed successfully, which may provide a basis for further exploration of function of IER3 in lung cancer.

Relationship between Cancer Stem Cell Marker CD133 and Cancer Germline Antigen Genes in NCI-H292 Lung Cancer Cells

  • Ko, Taek Yong;Kim, Jong In;Lee, Sang Ho
    • Journal of Chest Surgery
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    • v.53 no.1
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    • pp.22-27
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    • 2020
  • Background: Previous studies have shown that lung cancer stem cells express CD133 and that certain cancer stem cells express cancer germline antigens (CGAs). The transcriptional regulation of CD133 is complicated and poorly understood. We investigated CD133 and CGA expression in a non-small cell lung cancer cell line. Methods: The expression levels of CD133 and CGAs (MAGE-6, GAGE, SSX, and TRAG-3) were measured in an NCI-H292 lung cancer cell line. The methylation status of the CD133 gene promoter region was analyzed. The expression levels and promoter methylation statuses of CD133 and CGAs were confirmed by treatment with the demethylating agent 5-aza-2'-deoxycytidine (ADC). Results: After treatment with ADC, CD133 expression was no longer detected. MAGE-6 and TRAG-3 were detected before ADC treatment, while GAGE and SSX were not detected. ADC treatment upregulated MAGE-6 and TRAG-3 expression, while GAGE expression was still undetected after treatment, and only weak SSX expression was observed. GAGE expression was not correlated with expression of CD133, while the levels of expression of MAGE-6, TRAG-3, and SSX were inversely correlated with CD133 expression. Conclusion: These results showed that CD133 expression can be regulated by methylation. Thus, the demethylation of the CD133 promoter may compromise the treatment of lung cancer by inactivating cancer stem cells and/or activating CGAs.

Residential design on characteristics of formative expression in the contemporary residential architecture (현대 주거건축의 조형적 표현특성을 적용한 주거 공간 계획)

  • Park, Hae-Ri;Mo, Jeong-Hyun
    • Proceeding of Spring/Autumn Annual Conference of KHA
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    • 2009.11a
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    • pp.223-228
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    • 2009
  • The meaning of the characteristics of formative expression in contemporary residential architecture is flowing elements as important means to inbreathe emotional feeling of movement and as automatic and positive wats of expression. So the purpose of this study was to analyze the characteristics of formative expression in contemporary residential architecture, and to plan residential design linked with this kind of concept. For this study, analysis of case studies were used and the cases were selected in contemporary residential architecture which were represented having formative expressions since 2005. The major results found that the characteristics of formative expression were ex-center, reiteration, dispersibility, diagonality, plasticity, non-linearity, and deconstruction. In addition to that, we derived various methods expressing the formative characteristics, which were mass transformation, plan variation and composition element transformation. Summing these results up, the various characteristics of formative expression in contemporary residential architecture are understood as the results of the free modeling will of architects fulfilled with the spirit of inquiry and experiment to escape from the standard architecture in order to differentiate themselves.

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Interferon Induced Transmembrane Protein-1 Gene Expression is a Biomarker for Early Detection of Invasive Potential of Oral Squamous Cell Carcinomas

  • Ramanathan, Arvind
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.4
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    • pp.2297-2299
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    • 2016
  • Background: Early detection of malignant transformation with expression biomarkers has significant potential to improve the survival rate of patients as such biomarkers enable prediction of progression and assess sensitivity to chemotherapy. The expression of interferon inducible transmembrane protein 1 (IFITM1) has been associated with early invasion events in several carcinomas, including head and neck cancers, and hence has been proposed as a novel candidate biomarker. As the incidence of oral squamous cell carcinoma (OSCC) is highest in the Indian population, we sought to investigate: 1) the expression pattern of IFITM1 in OSCC tissue samples obtained from Indian patients of Dravidian origin; and 2) the possibility of using IFITM1 expression as a potential biomarker. Materials and Methods: Total RNA extracted from thirty eight OSCC biopsy samples was subjected to semi-quantitative RT-PCR with IFITM1 and GAPDH specific primers. Results: Of the thirty eight OSCC samples that were analyzed, IFITM1 overexpression was identified in fifteen (39%). Seven expressed a low level, while the remainder expressed high level of IFITM1. Conclusions: The overexpression of IFITM1 in OSCC samples indicates that IFITM1 may be explored for the possibility of use as a high confidence diagnostic biomarker in oral cancers. To the best of our knowledge, this is the first time that IFITM1 overexpression is being reported in Indian OSCC samples.

FHIT Gene Expression in Acute Lymphoblastic Leukemia and its Clinical Significance

  • Malak, Camelia A Abdel;Elghanam, Doaa M;Elbossaty, Walaa Fikry
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8197-8201
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    • 2016
  • Background: To investigate the expression of the fragile histidine triad (FHIT) gene in acute lymphoblastic leukemia and its clinical significance. Materials and Methods: The level of expressed FHIT mRNA in peripheral blood from 50 patients with acute lymphoblastic leukemia (ALL) and in 50 peripheral blood samples from healthy volunteers was measured via RT-PCR. Correlation analyses between FHIT gene expression and clinical characteristics (gender, age, white blood count, immunophenotype of acute lymphoblastic leukemia and percentage of blast cells) of the patients were performed. Results: The FHIT gene was expressed at $2.49{\pm}7.37$ of ALL patients against $14.4{\pm}17.9$ in the healthy volunteers. The difference in the expression levels between ALL patients and healthy volunteers was statistically significant. The rate of gene expression did not significantly vary with immunophenotype subtypes. Gene expression was also found to be correlated with increase of total leukocyte and decrease in platelets, but not with age, gender, immunophenotyping or percentage of blast cells. Conclusions: FHIT gene expression is low in acute lymphoblastic leukemia and could be a useful marker to monitor minimal residual disease. This gene is also a candidate target for the immunotherapy of acute lymphoblastic leukemia.

Effects of Sohaphyang-won on the Gene Expression in a Hypoxic Model of Cultured Rat Cortical Cells (배양한 흰쥐 대뇌세포의 저산소증 모델에서 소합향원이 유전자 표현에 미치는 영향)

  • 백진원;이영효;김완식;정승현;신길조;이원철
    • The Journal of Korean Medicine
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    • v.25 no.2
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    • pp.127-137
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    • 2004
  • Objectives : The purpose of this investigation was to evaluate the effects of Sohaphyang-won (SH) on the alteration in gene expression in a hypoxia model using cultured rat cortical cells. Methods : E18 rat cortical cells were grown in neurobasal medium containing B27 supplement. On 12 DIV, SH was added ($20\mu\textrm{g}/ml$) to the culture media for 24 hrs. On 14 DIV, cells were given a hypoxic insult (2% O2/5% CO2, $37^{\circ}C$, 3 hrs), returned to normoxia and cultured for another 24 hrs. Total RNA was prepared from SH-untreated (control) and -treated cultures and alteration in gene expression was analyzed by microarray using rat 5K-TwinChips. Results : Effects on some of the genes whose functions are implicated in neural viability are as follows: 1) For most of the genes altered in expression, the global M values were between -05 to +0.5, Among these, 1517 genes were increased in their expression by more than global M +0.1, while 1480 genes were decreased by more than global M -0.1. 2) The expression of apoptosis-related genes such as Bad (global M =0.35), tumor protein p53 (T53) (global M =0.28) were increased, while v-akt murine thymoma viral oncogene homolog 1 (Akt1) was decreased. 3) The expression of hemoglobin alpha 1 (probably neuroglobin) was increased by about 3.2-fold (global M =1.7). 4) The expression of antioxidation-related catalase gene was increased (global M =0.26). 5) The expression of PKCzeta (prkcz), an upstream kinase of MAPK, was increased (global M =0.29). 6) The expression of retinoic acid receptor alpha (RAR), which may regulate transcription in hypoxic stress, was increased (global M =10.27). Conclusions : In summary, the microarray data suggest that SH doesn't increase the expression of oxygen capture-, anti-oxidation- and 'response to stress' -related genes but decreases some anti-apoptosis genes which would help protect the hypoxic cells from apoptosis.

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Patterns of Aquaporin 7 Expression in Normal Follicles and Follicular Cyst Follicles of Hanwoo (한우의 정상 난포와 난포낭종 난포에서 Aquaporin7 발현 양상)

  • Kim, Chang-Woon;Han, Sunkyu;Choe, Changyong
    • Journal of Embryo Transfer
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    • v.30 no.1
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    • pp.17-21
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    • 2015
  • Alteration in ion channel or transporter expression levels affects cell volume which is produced by movement of water and ion across the plasma membrane. In particular, aquaporin (AQP) channels among ion channels play a crucial role in movement of water across the cell membrane. This study was performed to identify whether AQP expression is changed in bovine follicular cystic follicles using microarray, RT-PCR and Western blotting analyses. In microarray data, AQP4 expression was decreased, whereas AQP7 was increased in cystic follicles. Additional experiments were focused on the AQP7 expression increased in cystic follicles. The microarray data was confirmed by semi-quantitative polymerase chain reaction (PCR) and Western blot analysis. AQP7 mRNA and protein expressions were significantly increased in the cystic follicles (p<0.05). Application of estrogen ($10{\mu}g/ml$) to bovine ovarian cells showed a trend of increase in AQP7 expression. From these results, we suggest that the increase in AQP7 expression in cystic follicles may play an important role in movement of water in bovine ovary. In addition, AQP7, a aquaglyceroporin permeating water and glycerol, could be a good target in development of methods for the cryopreservation of bovine ovary.

Analysis and Proposal of Abstractive Expression of 3D Animation (3D 애니메이션의 추상적 표현에 관한 분석 및 제안)

  • Park, Sung-Dae;Jung, Yee-Ji
    • Cartoon and Animation Studies
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    • s.37
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    • pp.1-24
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    • 2014
  • The advancement of computer technology has blurred the border in image expression between live-action films and 3D animation to an indistinguishable extent. Despite the ever-evolving image expression, it has been found that animation properly incorporating abstractive expression receives more positive responses from the audience than the animation closely akin to live-action films. This indicates that animation-specific abstractive expression can exert more significant effects than that of live-action expression as proved in Dr. Mori Masahiro's 'Uncanny Valley.' Also, such effects apply only to characters, whilst the technology capable of live-action expression is acknowledged only for the background of 3D animation. Without doubt, animation has its own infinite world of expression, and thus more effective methods for its expression can no longer be considered a matter of technology. The present study analyzes the abstractive expression of 3D animation in terms of characters and backgrounds and explores a more effective method to apply the current technology of image expression to 3D animation with a view to proposing a direction for more appropriate application of 3D animation technology to future animation.