• Title/Summary/Keyword: FAS1

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Comparison between Doxorubicin and Anti-Fas Antibody induced poptosis in Promyelocytic Leukemia Cell Line HL-60 (전골수성 백혈병 세포주 HL-60에 대한 Doxorubicin 유발성 Apoptosis와 Anti-Fas 항체 유발성 Apoptosis의 비교)

  • 윤경식;설지연;오현정;이광수;이원규;정성철
    • Biomolecules & Therapeutics
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    • v.7 no.1
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    • pp.22-28
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    • 1999
  • Induction of apoptosis is considered to be the underlying mechanism that accounts for the efficiency of chemotherapeutic drugs. It has recently been proposed that doxorubicin (DOX) can induce apoptosis in human leukemic cells via the Fas/Fas Ligand (FasL) system. Comparison of Fas and FasL mRNA expression between drug- and anti-Fas antibody(Fas-Ab)- induced apoptosis was analyzed for examining the role of Fas/FasL system in the mediation of drug-induced apoptosis. After HL-60 cells were routinely cultured, MTT assay was performed for cytotoxicity test. Giemsa staining was carried out to monitor the apoptosis morphologically. By semiquantitative RT-PCR analysis, the expression of Fas and FasL at 4, 10, 24 hours was determined after DOX and Fas-Ab treatment. Dose-dependent cytotoxicity was induced by DOX-treatment, while Fas-Ab treatment showed the similar dose-dependent pattern but the cytotoxicity is not reached at LD$_{50}$ at 100 ng/ml concentration of Fas-Ab. In the 10ng/m1 DOX and 10ng/m1 Fas-Ab treated group, typical apoptotic cell morphology was shown such as fragmented nuclei and cell membrane budding in the Giemsa-stained slide. Fas mRNA expression was not changed significantly in the both groups. But, FasL mRNA expression was induced significantly at initial period of apoptosis. In this study, Fas/FasL interaction assumed to be involved in drug-induced apoptosis.s.

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Expression and Preparation of Periostin FAS1 Domains for NMR Structure Determination

  • Yun, Hyosuk;Kim, Jae Il;Lee, Chul Won
    • Journal of the Korean Magnetic Resonance Society
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    • v.20 no.1
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    • pp.17-21
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    • 2016
  • Periostin, a component of extracellular matrix (ECM) protein, is produced and secreted by the fibroblasts that are involved in chronic allergic inflammation diseases and various types of human cancers. Periostin protein is composed of multiple domains including four FAS1 domains which play important roles in cell adhesion and tumor metastasis by interacting with integrins. In spite of their important biological role, the structural information of periosin FAS1 domains was not revealed yet. Recently we systemically prepared various constructs of the FAS1 domains and tried to express them in E. coli. Of them, only single FAS1-II and -IV domains were highly soluble. Circular dichroism (CD) and nuclear magnetic resonance (NMR) studies revealed that the FAS1-IV domain might be suitable for three-dimensional structure determination using NMR spectroscopy.

Fas/FasL expression in the hippocampus of neonatal rat brains follwing hypoxic-ischemic injury (저산소성 허혈성 손상을 받은 신생 흰쥐 뇌 해마에서 Fas와 FasL 단백 발현)

  • Chang, Young Pyo;Kim, Myeung Ju;Lee, Young Il;Im, Ik Je;Cho, Jae Ju;Kim, Jong Wan;Yeo, Sung Moon
    • Clinical and Experimental Pediatrics
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    • v.49 no.2
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    • pp.198-202
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    • 2006
  • Purpose : Fas is a cell surface receptor that transduces apoptotic death signals. Interaction of extracelluar domain of Fas with Fas ligand(FasL) triggers the apoptotic process in many diseases. We investigated the expression of Fas and FasL in the hippocampus of 7-day-old newborn rat brains following hypoxia-ischemia injury. Methods : The 7-days-old newborn rats were exposed to 8 percent oxygen for two hours after the ligation of right common carotid arteries. The newborn rats were killed and their brains were removed at 12, 14 and 48 hours after hypoxic-ischemic injury. The expressions of Fas and FasL of the right hippocampus were observed by western blotting and immunofluorescent staining. Results : Fas and FasL were strongly expressed in the right hippocampus ipsilateral to the ligation of the common carotid artery by western blotting at 12 hours following hypoxic-ischemic injury, and then slowly decreased. The immunofluorescent expressions of Fas and FasL strongly increased in the CA1 area of the right hippocampus at 12 and 24 hours following hypoxic-ischemic injury. The immunofluorescent expression of Fas decreased at 48 hours, but the expression of FasL persisted strongly at 48 hours following hypoxic-ischemic injury. Conclusion : The interaction of Fas with FasL on the cell surface may be involved in neuronal injury following hypoxic-ischemic injury in the developing brain.

Upregulation of Fas in epithelial ovarian cancer reverses the development of resistance to Cisplatin

  • Fan, Yang;Wang, Long;Han, Xuechuan;Liu, Xueqin;Ma, Hongyun;Ding, Yonghui
    • BMB Reports
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    • v.48 no.1
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    • pp.30-35
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    • 2015
  • This study was to investigate the role of Fas in the development of Cisplatin-resistant ovarian cancer. On the cellular level, Fas expression was significantly reduced in Cisplatin resistant A2780 (A2780/CP) cells compared with A2780 cells. Fas silence with siRNA would promote tumor cell lines proliferation, facilitate tumor cell cycle transition of G1/S, prevent cell apoptosis, and promote cell migration. Expression of drug resistance gene was negatively correlated to Fas. In nude mice metastasis model of human ovarian carcinoma by subcutaneous transplantation, after Ad-Fas injected intratumorly, we found that upregulation of Fas could inhibit transplantation tumor tissue growth and reduce the expression of drug resistance gene. Our results indicated that upregulation of Fas in epithelial ovarian cancer reversed the development of resistance to Cisplatin. In conclusion, our findings suggested that Fas might act as a promising therapeutic target for improvement of the sensibility to Cisplatin in ovarian cancer.

Apo-1/Fas (CD95) Gene Polymorphism in Korean Hepatocellular Carcinoma Patients

  • Kim, Sung-Soo;Hong, Seung-Jae;Ahn, Yun-Gul;Kim, Bong-Seog;Yuh, Young-Jin;Han, Kye-Young;Lee, Hee-Jae;Chung, Joo-Ho;Yim, Sung-Vin;Cho, Jae-Young;Park, Yeon-Hee
    • The Korean Journal of Physiology and Pharmacology
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    • v.7 no.1
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    • pp.29-31
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    • 2003
  • It is well known that different expression of Apo-1/Fas (CD95) plays important roles in various tumors and hepatocellular carcinoma (HCC) pathogenesis. Apo-1/Fas mediated apoptosis is one of the important pathways of apoptosis and is known to mediate apoptotic cell death by fas ligand (FasL). To examine the possible relationship between Apo-1/Fas gene polymorphism and HCC susceptibility, MvaI restriction fragment length polymorphism (RFLP) of Apo-1/Fas gene was examined in 94 Korean HCC patients and 240 control subjects. No statistically significant difference in the genotypic distribution and allelic frequencies was found between the control and the HCC. It is, therefore, concluded that Apo-1/Fas gene polymorphism is not associated with HCC susceptibility. Further studies are needed in order to clarify the relationships between genotypes of Apo-1/Fas gene and HCC pathogenesis.

Apoptosis in Human Corneal Epithelial cells induced by Exhausted Medium (Exhausted Medium에 의한 각막상피 세포의 세포고사 유도)

  • Kim, Jae-Min
    • Journal of Korean Ophthalmic Optics Society
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    • v.5 no.1
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    • pp.83-87
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    • 2000
  • To investigate exhausted-medium-induced apoptosis in human corneal epithelial(HCE) cells, this study was performed DNA gel electrophoresis, M30 CytoDEATH staining and FAS-FAS ligand ELISA. SV-40 transfected cells were grown to confluency in culture for 7days. The supernatant was harvested and filtered with $0.22{\mu}m$ filter paper. Fresh HCE cells were exposed to the filtered exhausted medium for 1~2 days. Apoptotic cells were prepared for DNA extraction and run the agarose gel for DNA ladder pattern. M30 CytoDEATH was used a tool for easy and reliable determination of very early apoptosis in HCE cells. The control and exhausted medium were assayed for soluble FAS/FAS ligand protein by ELISA. HCE cells exposed to exhausted medium showed a typical DNA ladder pattern. Sporadic M30 CytoDEATH positive cells were detected among HCE cells exposed to exhausted medium. Soluble FAS/FAS ligand levels were not elevated in the exhausted medium compared to the fresh medium control. This study suggests that possible mechanism of exhausted medium induced apoptosis does not include the FAS-FAS ligand system.

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Objective and Subjective Socioeconomic Position and Current Smoking Among Korean Adolescents

  • Ko, Min Jung;Lee, Eun Young;Kim, Kirang
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.20
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    • pp.8877-8881
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    • 2014
  • Background: Despite social gradients in adult smoking, the effects of socioeconomic position (SEP) on adolescent smoking is not well understood. This study examined effects of subjective SEP as well as the objective SEP on smoking among Korean adolescents. Materials and Methods: Data were obtained from the 2012 Korea Youth Risk Behavior Web-based Survey, a nationally representative sample of middle and high school students (38,221 boys; 35,965 girls). SEP was assessed by the Family Affluence Scale (FAS) and the self-rated household economic status. Relationships between SEP and smoking were analyzed by multivariate logistic regression. Results: The low perceived SEP for either the high or low FAS grade was related to an elevated likelihood of smoking in both genders. A significantly higher risk of smoking was found in boys of low perceived SEP in middle school (odds ratio [OR] 1.50; 95% confidence interval [CI] 1.28-1.77 for high FAS, OR 1.55; 95% CI 1.21-1.98 for low FAS), and of low perceived SEP and high FAS in high school (OR 1.13; 95% CI 1.02-1.26). Among girls, an elevated risk of smoking was observed in middle school group with low perceived SEP and low FAS (OR 2.01; 95% CI 1.44-2.79) and in the high school group of low perceived SEP, regardless of FAS level (OR 1.34; 95% CI 1.14-1.57 for high FAS, OR 1.31; 95% CI 1.04-1.65 for low FAS). Conclusions: The relationship of subjectively perceived SEP with smoking is as important as objective SEP and more significant in Korean high school adolescents.

Apoptosis and Expression of Fas nnd Fas Lignnd Genes in Mouse Ovary (생쥐 난소내 Fas 및 Fas Ligand의 발현과 세포자연사)

  • Yoon Jeong-Mi;Yoon Hyunsook;Yang Hyunwon;Kim Seikwang;Cho Dong Jae;Yoon Yong-Dal
    • Development and Reproduction
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    • v.7 no.1
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    • pp.15-22
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    • 2003
  • The Fas antigen (Fas) as a cell-surface receptor protein which mediates apoptosis-inducing signals plays an important role in the immune system. Expression of Fas mRNA is detected not only in lymphoid organs but also in the nonlymphoid organs. In the ovary, most of the follicles is known to undergo atreisa through apoptosis. However, the exact mechanism of atresia was not elucidated yet. Therefore, the purposes of the present study were to investigate the expression of Fas and Fas ligand in mouse ovary and to clarify the relationship between expression of Fas and Fas ligand and atresia of follicle. The result of RT-PCR demonstrated that Fas and Fas ligand mRNA was expressed in ovary, especially granulosa cells and oocytes. The immunohistochemistry showed that the granulosa cells and oocytes in growing follicles were stained for Fas and Fas ligand, but primordial follicles were not. Furthermore, Fas and Fas ligand were intensively stained in the atretic follicles As results of TUNEL staining to detect apoptotic cells in the ovaries, the number of TUNEL-positive (apoptotic) granulosa cells and oocytes increased in the atretic follicles compared to the healthy normal follicles. These results demonstrate that there is the positive relationship between expression of Fas and Fas ligand in granulosa cells and oocyies and apoptosis of them leading to atresia of follicles. It suggests that expression of Fas and Fas ligand could be associated with atresia of follicles in mouse ovary.

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Effects of FasL Expression in Oral Squamous Cell Cancer

  • Fang, Li;Sun, Lin;Hu, Fang-Fang;Chen, Qiao-Er
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.1
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    • pp.281-285
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    • 2013
  • Purpose: To probe the role of FasL in cell apoptosis in oral squamous cell carcinomas (OSCCs). Methods: The expression of Fas/FasL was assessed in 10 cases of normal oral epithelium, 38 cases of OSCC and tumor infiltrating lymphocytes (TIL), and 11 cases of metastatic lymph nodes by immunohistochemistry. Apoptosis of tumor cells and TIL was detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL). FasL-induction of T cell apoptosis was tested by co-culture assay in vitro with SCC-9 and Jurkat T cells. Results: The 10 cases of normal oral epithelium all demonstrated extensive expression of Fas, the positive rate being largely down-regulated in OSCC (21/38) (P<0.05) compared to the normal (10/10). At the same time, the positive rate of FasL significantly increased in OSCC (P<0.05) especially those with lymph node metastasis (P<0.05). The positive rates of Fas in well and middle differentiated OSCC were higher than those in poor differentiated OSCC (P<0.05). The AI of tumor cells in Fas-positive OSCC was remarkably higher than that in Fas-negative OSCC (P<0.01), with a positive correlation between Fas expression and cell differentiation as well as apoptosis (r=0.68, P<0.01). The AI of tumor cells in FasL positive OSCC was remarkably lower than that in control while the AI of TIL was higher than in FasL negative OSCC (P<0.05). The AI of tumor cells reversely correlated with that of TIL (r = -0. 72, P<0.05). It was found that SCC-9 cells expressing functional FasL could induce apoptosis of Jurkat cells as demonstrated by co-culture assays. As a conclusion, it is evident that OSCC cells expressing FasL can induce apoptosis in Fas-expressing T cells. Conclusions: In progression of OSCC, expression of the Fas/FasL changes significantly. The results suggest that FasL is a mediator of immune privilege in OSCC and may serve as an marker for predicting malignant change in oral tissues.

Apo-1/Fas (CD95) Gene Polymorphism in Korean Knee Osteoarthritis Patients (한국인 무릎 골 관절염 환자들의 임상 양상과 Apo-1/Fas (CD95) 유전자 다형성과의 상관관계에 관한 연구)

  • Hong, Seung-Jae;Yang, Hyung-In;Yim, Sung-Vin;Chung, Joo-Ho;Jung, Young-Ok;Kim, Ho-Youn
    • IMMUNE NETWORK
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    • v.3 no.2
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    • pp.145-149
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    • 2003
  • Background: Apoptosis has been implicated in pathogenesis of various disease. Apo-1/Fas (CD95) is one of the main pathway of apoptosis. To examine the possible relationship between Apo-1/Fas (CD95) and primary knee osteoarthritis, MvaI restriction length polymorphism (RFLP) in human Apo-1/Fas (CD95) gene was assessed. Methods: Genotype and allele frequencies in promoter region in the Apo-1/Fas (CD95) gene were studied by PCR-RFLP in 226 Korean controls and 148 Korean patients with primary knee osteoarthritis. Results: No statistically significant difference in the genotypic distribution and allelic frequencies was found between the control and the knee oateoarthritis patients. But in the severe grade (grade 3, 4) Kellgren-Lawrence score patients, the frequency of $MvaI^*1$ (G) allele was significantly decreased (P=0.0392) and the of $MvaI^*2$ (A) allele frequency was significantly increased (P=0.0473) compared to the normal controls. Conclusion: Apo-1/Fas (CD95) gene polymorphism is a part a determinant factor of severity in knee osteoarthritis, the patients with $MvaI^*2$ (A) allele is more severe radiologic progression. Further substantiation studies are needed in larger patient samples and various other apoptosis related genes to elucidate the mechanism of osteoarthritis, including the Fas ligand gene analysis.