• Biomolecules & Therapeutics
• /
• 제7권1호
• /
• pp.22-28
• /
• 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.

• 한국자기공명학회논문지
• /
• 제20권1호
• /
• pp.17-21
• /
• 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.

• Clinical and Experimental Pediatrics
• /
• 제49권2호
• /
• pp.198-202
• /
• 2006

목 적 : Fas는 세포표면 수용체로 세포사멸 신호를 전도한다. 많은 질환에서 세포표면의 Fas가 Fas ligand(FasL)와 결합하여 세포사멸 과정을 유발하게 된다. 연구자들은 7일된 신생 흰쥐에 저산소성 허혈성 손상을 유발한 후 뇌 해마에서 Fas와 FasL의 발현을 관찰하고자 하였다. 방 법 : 7일된 신생 흰쥐를 오른쪽 총 경동맥 영구 결찰 후 8% 산소에 2시간 노출시켰다. 저산소성 허혈성 손상 후 12, 24, 48시간에 뇌를 적출 냉동 보관하였다. Western blotting 방법과 면역형광염색 방법으로 냉동 보관된 뇌의 경동맥을 결찰한 오른 쪽 해마에서 Fas와 FasL의 발현을 관찰하였다. 결 과 : Fas와 FasL의 발현은 저산소성 허혈성 손상 후 12시간에 경동맥이 결찰된 오른쪽 해마에서 크게 증가하고 이후 감소하는 것을 western blotting 방법에 의해 관찰하였다. Fas와 FasL의 면역형광발현은 오른쪽 해마의 CA1 영역에서 손상 후 12시간과 24시간에 대조군에 비해 증가하였다. Fas의 면역형광 발현은 손상 후 48시간에 감소하였으나 FasL의 면역형광발현은 손상 후 48시간에도 지속되었다. 결 론 : 세포표면에서 Fas와 FasL의 발현과 그들의 결합은 저산소성 허혈성 손상을 받은 미성숙 뇌의 신경세포 손상에 기여할 것으로 추측되었다.

• BMB Reports
• /
• 제48권1호
• /
• pp.30-35
• /
• 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.

• The Korean Journal of Physiology and Pharmacology
• /
• 제7권1호
• /
• pp.29-31
• /
• 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.

• 한국안광학회지
• /
• 제5권1호
• /
• pp.83-87
• /
• 2000

본 연구는 각막상피가 저절로 탈락해 나가는 과정을 이해하기 위해 사람의 각막상피 세포를 배양하여 배지의 영양이 고갈될 때까지 약 7일 정도 배지를 교환하지 않고 계속 배양하여 세포고사를 조사하였다. 영양이 고갈된 배지인 Exhausted Medium을 여과하여 새로운 각막 상피세포에 넣어 2일 정도 배양을 계속하였다. Exhausted Medium에서 배양된 세포를 수확하여 세포고사를 검정하기 위해 Agarose gel electrophoresis로 DNA 단편을 확인하고 세포고사를 초기에 감지할 수 있는 방법인 M30 $CytoDEATH^*$. Fluorescein으로 확인하였다. 또한 Exhausted Medium에 의한 세포고사의 유발 경로를 조사하기 위해 사이토카인에 의한 유도인자 중 가장 많이 알려진 FAS나 FAS Ligand에 의한 유발여부를 soluble FAS and FAS Ligand에 대한 sandwich ELISA로 조사하였다. 그 결과 전형적인 DNA Ladder패턴을 보였으며 M30 $CytoDEATH^*$. Fluorescein에도 선명하게 염색되어 세포고사를 확인할 수 있었다. 또한 soluble FAS and FAS Ligand ELISA Kit로 Exhausted Medium이 FAS와 관련이 있는지를 조사한 결과 거의 무관한 것으로 나타났다. 본 연구의 결과는 영양 부족상태의 배지는 각막상피 세포가 세포고사를 거쳐 소모되는데 FAS and FAS Ligand system과는 무관한 것으로 나타났다.

• Asian Pacific Journal of Cancer Prevention
• /
• 제15권20호
• /
• pp.8877-8881
• /
• 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.

• 한국발생생물학회지:발생과생식
• /
• 제7권1호
• /
• pp.15-22
• /
• 2003

Fas는 세포자연사를 유도하는 세포 표면 수용체 단백질로서 면역계에서 중요한 역할을 한다. 이러한 Fas mRNA는 림프조직뿐만 아니 라 비 림프조직에서도 발현한다. 한편 대다수의 난포들은 세포자연사와 연관된 기 작을 통해 난포폐쇄로 진행되는 것으로 알려지고 있으나, 난포폐쇄에 관한 기작은 아직 규명되지 않았다. 따라서 본 연구의 목적은 먼저 생쥐의 난소의 과립세포와 난자에서 Fas와 Fas ligand의 발현 여부를 알아보고, Fas와 Fas ligand발현과 난포폐쇄와의 상관 관계를 알아보고자 하였다. RT-PCR을 수행한 결과, 난소내 과립세포와 난자에서 Fas와 Fas ligand mRNA가 모두 발현하는 것을 확인할 수 있었다. 면역조직화학적 염색 결과 또한, 원시 난포를 제외한 모든 발달 중인 난포의 과립세포와 난자들에서 Fas와 Fas ligand가 검출되는 것을 확인하였고, 특히 폐쇄를 보이는 난포에서 강한 발현을 보였다. 한편, 난소에서 세포자연사를 확인하기 위해 TUNEL 염색을 수행한 결과, TUNEL 양성을 보이는 과립세포와 난자의 수는 건강한 정상 난포에 비해 폐쇄난포에서 증가하는 것을 관찰할 수 있었다. 이상의 결과들은 난소내 과립세포와 난자에서 Fas와 Fas ligand 발현과 난포폐쇄와 상관 관계가 있음을 보여주고 있으며, 이러한 Fas와 Fas ligand가 생쥐 난소내 난포폐쇄 유발에 관여하고 있을 것으로 사료된다

• Asian Pacific Journal of Cancer Prevention
• /
• 제14권1호
• /
• pp.281-285
• /
• 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.

• IMMUNE NETWORK
• /
• 제3권2호
• /
• pp.145-149
• /
• 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.