miR-21 influences growth of glioma cells by targeting FasL gene / 中国病理生理杂志

AIM:ToinvestigatetheregulationofmiR-21onFasLexpressionanditseffectonthegrowthand apoptosis in glioma cells , and to evaluate the molecular mechanism .METHODS:Differential expression levels of miR-21 in human glioma U251 cells were achieved by transfecting with miR-21 mimics, miR-21 inhibitor or scramble .The viability and apoptosis of U251 cells were detected by CCK-8 assay and flow cytometry with Annexin V-FITC/PI double staining. The recombination vector pmirGLO-FasL was constructed .Dual-luciferase reporter experiment was performed to validate the target genes of miR-21.The expression vector pcDNA3.1-FasL was also constructed , and the biological activity and regula-tory role of miR-21 in U251 cell apoptosis were analyzed by a restore experiment .RESULTS:Exogenous overexpression of miR-21 increased the viability and decreased the apoptosis of U 251 cells ( P<0.05 ) , while miR-21 inhibitors generated the opposite results (P<0.05).Dual-luciferase reporter assay and restore experiment revealed that miR-21 negatively reg-ulated the expression of FasL gene which was regarded as the target gene , thus decreasing the apoptosis of U 251 cells. CONCLUSION:miR-21 increases the viability of glioma U251 cells, in which FasL may be one of the target genes .

Association of Fas, Fas-ligand gene polymorphisms with bone mineral desity and bone resposiveness to hormone therapy in postmenopausal women / 대한산부인과학회지

OBJECTIVE: To investigate the relationship between Fas gene & Fas-ligand gene polymorphisms, and bone mineral density (BMD) after hormone therapy (HT) in postmenopausal women. METHODS: Restriction fragment length polymorphisms at the Fas A670G, G1377A gene site and Fas-ligand C843T, IVS3nt169 (T/delT) gene site and BMD at the lumbar spine and proximal femur were analyzed in 229 postmenopausal women receiving sequential HT for 1 year. BMD were measured by DEXA. The subjects were divided in normal, osteopenic and osteoporotic on the basis of the T-score values according to the classification of the World Health Organization (WHO). RESULTS: After adjusting for potential confounding factors such as age, BMI, and menopause duration, A670G polymorphism was significantly associated with BMD at the lumbar spine, the femur neck and trochanter in osteopenic and osteoporotic groups, and G1377A polymorphism was significantly associated with BMD at lumbar spine and the femur neck in osteopenic group. C843T polymorphism was significantly associated with BMD at lumbar spine and ward triangle in osteoporotic group, IVS3nt169 (T/delT) was not associated with BMD. In osteoporotic group after HT in postmenopausal women, A670G polymorphism A/A, G1377A polymorphism G/G, C843T polymorphism T/T were associated with significant annual bone mineral density change, compared with other polymorphism at the same gene. CONCLUSION: These findings suggest that Fas, Fas-ligand gene polymorphisms may be an important contributor to the variation of BMD among postmenopausal women. and that a specific Fas, Fas-ligand polymorphisms are associated with significant BMD change in postmenopausal women after HT.

Telomerase activity and Expression of MIB-1, Fas and Fas Ligand in Placentas from Women with and without Intrauterine Growth Retardation

BACKGROUND: The placenta from a pregnancy that is complicated by intrauterine growth retardation (IUGR) tends to be smaller than that from a normal pregnancy. To investigate this difference, we analyzed the telomerase activity, the proliferative activity and the mRNA levels of apoptosis mediators in placentas. METHODS: In 20 placentas from normal third-trimester pregnancies and 22 placentas form pregnancies that were complicated by IUGR, the telomerase activity was detected by a telomeric repeat amplification protocol assay. The proliferative activity was assessed by immunohistochemical staining using the MIB-1 monoclonal antibody. The expression of the apoptosis mediator was evaluated by semi-quantitative reverse transcription-polymerase chain reactions for fas and fas ligand. RESULTS: Telomerase activity was detected in 2 (10%) of 20 normal placentas, whereas it was not observed in all tested 13 placentas that were associated with IUGR. The proliferative activity was significantly low in the placentas that were associated with IUGR (7.44+/-2.96%), compared with the normal placentas (11.0+/-3.48%, p=0.002). There was no statistically significant difference in the mRNA levels of fas or fas ligand between two groups. CONCLUSIONS: Low telomerase and proliferative activities in the placenta may play a role in the pathogenesis of IUGR.

Expression of Gastrin,Somatostatin,Proliferating Cell Nuclear Antigen and Fas-Ligand in Mucosa of Gastric Antrum in Children with Chronic Gastritis / 实用儿科临床杂志

Objective To investigate the expression of immunohistochemistry of gastrin(GAS),somatostatin(SS),proliferating cell nuclear antigen(PCNA) and Fas-ligand(Fas-L) in the sinus ventriculi of children with pediatric gastritis and to explore the significance of their expression in the pathogenesis of pediatric chronic gastritis.Methods Fifty cases of the sinus ventriculi mucosa samples were enrolled in 3 groups:chronic gastritis,helicobacter pylori(Hp) positive(group A,n=20);chronic gastritis,Hp negative(group B,n=19);control group,normal sinus ventriculi mucosa,Hp negative(group C,n=11).Immunohistochemistry En Vision were carried out including GAS,SS,PCNA and Fas-L.Results In the expression of GAS and SS,the values of group A and B were comparatively higher than those of group C,but there was no significant difference among them in statistics.In the expression of PCNA,the value of group A was comparatively higher and that of group B.The value difference between 2 groups was significant(P=0.019);in the expression of Fas-L,no significant difference was found among these 3 groups.Conclusions Expressions of GAS and SS both increase in children with chronic gastritis and maybe the increase of GAS and SS play a role in the pathogenesis of pediatric chronic gastritis;Hp infection promotes the multiplication of the sinus ventriculi membrana mucosa epithelium cell in pediatric chronic gastritis.

Expression of Fas, Fas-ligand, Bcl-2 and Bad with Maturation of Human Ovarian Follicle / 대한산부인과학회잡지

Human ovarian follicles reduce rapidly in number throughout fetal and adult life. Throughout the menstrual cycles, primordial follicles grow into mature follicles and then ovulate to form corpus luteum. Apoptosis has been implicated in several events that occur during the process of follicular growth, atresia and the regression of the corpus luteum. By the use of immunohistochemistry, we clarified the involvement of apoptosis in the human ovary during follicular growth, regression and atresia by investigating the expression of Fas, Fas-ligand, Bcl-2 and Bad in primordial follicles, primary follicles and mature follicles. Fas immunostaining was present in primordial oocytes, both oocytes and granulosa cells of primary follicles, preantral follicles and all follicular cells of mature follicles. Fas-ligand and Bad immunostaining patterns were similar to those of Fas except for theca cells. Bcl-2 immunostaining was present in both oocytes and granulosa cells of primary, preantral and mature follicles. In corpus luteum, Fas, Fas-ligand, Bcl-2 and Bad immunostaining were observed and decreased in the regressing corpus luteum. In postmenopausal ovary, Fas, Fas-ligand, Bcl-2 and Bad immunostaining were entirely negative. Bad immunostaining was observed but Bcl-2 was not in atretic follicle. These results suggest that Fas, Fas-ligand, Bcl-2 and Bad may play important roles in human ovary during follicular growth, regression and atresia simultaneously. Further studies should be required to elucidate the underlying mechanism and apoptosis of the disease associated with normal and abnormal ovarian aging.

The Expression of Peforin, Fas-ligand, and Granzyme B in Peripheral Blood Lymphocytes of Renal Allograft Recipients / 대한신장학회잡지

BACKGROUND: Previous findings demonstrated that the expression of cytotoxic effector molecules is increased in acute rejection of renal allografts. In the present study, we serially examined the gene expression of perforin, granzyme B and Fas ligand(FasL) in peripheral blood lymphocytes(PBLs) of renal allograft recipients to assess the potential of their expression as a marker of acute rejection. METHODS: PBLs were isolated from blood samples taken on days 2, 4, 6, 8, 10 and 12 after transplantation. Competitive PCR was performed to evaluate the abundance of mRNA of perforin, granzyme B and FasL. The mean value of each molecule plus 2 SD for the control group was set as a discriminatory level. RESULTS: When all measured samples were compared, perforin expression was significantly higher in patients with acute rejection than in the control group(1.84+/-3.01 versus 0.71+/-0.48, p=0.01). The percentage of perforin expression exceeding the discriminatory level was also significantly higher in patients with acute rejection(p=0.0003). Five patients in the rejection group(5/7, 71.4%) showed perforin expression exceeding the discriminatory level, while only 1 patient in the control group did so(1/8, 12.5%)(p= 0.02). Perforin expression of days 0 and 1 of rejection crisis was the highest over the study period. No consistent pattern of granzyme B and FasL expression was identified in relation to rejection crisis. CONCLUSION: Gene expression of perforin by PBLs was upregulated in accordance with acute rejection, thus offering the possibility that it may be utilized as a marker of acute rejection.

The Study on Apoptosis and Expression of Fas, Fas-ligand, Bax, and Bcl-2 in Human Fragmented Embryos / 대한불임학회지

OBJECTIVE: The present study was performed to investigate whether apoptosis occur in human embryos by annexin staining and detect the expression of Fas, Fas-ligand (FasL), Bax, and Bcl-2 in human fragmented embryos derived from IVF-ET by immunofluorescence and Western blot analysis. MATERIALS AND METHODS: Using annexin staining, immunofluorescence and Western blot analysis on normal and fragmented embryos, we were able to detect apoptotsis and apoptotic gene products in fragmented embryos. RESULTS: Phosphatidylserine (PS) translocation, the marker for apoptosis, were detected frequently in fragmented embryos. Bcl-2 and Bax protein were detected in both fragmented and non-fragmented embryos. When fragmented embryos compared to normal embryos, immunofluorescent intensity of Bcl-2 tended to be lower in fragmented embryos. Bax gene expression increased in the fragmented embryos compared to the normal embryos. This result supports a model in which the molar ratio of Bcl-2 to Bax determines whether apoptosis induced or inhibited in human embryo. Fas was highly expressed in human preimplantation embryos but not FasL. It suggests that embryo may undergo apoptosis by binding with FasL produced by follicular or immune cells. CONCLUSION: The over expression of Bax and Fas will trigger apoptosis to lead embryo fragmentation and change embryo to be nonviable.

The Immunologic Expression of Apoptosis Related Proteins and Apoptotic Cells in Human Ovarian Follicles / 대한불임학회지

OBJECTIVE: To investigate the expression of apoptosis related proteins and apoptotic cells on the human ovarian follicles. MATERIALS AND METHODS: Thirty five Formalin-fixed paraffin-embedded human ovarian tissue blocks were selected from the surgical pathology files of the department of pathology, College of Medicine, Yonsei University, for the period from 1996 to 1998. All specimen were from premenopausal women aged from 32~45. Ovarian tissues were collected from the patients performing hysterectomy for benign uterine diseases. Immunohistochemical staining was performed for the detection of DNA fragmented cell, Bcl-2, Bax, Fas and Fas-ligand. RESULTS: Bcl-2 and bax were not expressed on the surrounding cells and oocyte of the primary, primordial and preantral follicles. Fas and Fas-ligand (Fas-L) were not expressed on the surrounding cells on the primordial and primary follicles. But expressed on the surrounding granulosa cells and oocyte in the primordial and primary follicles. In the healthy follicles, Bcl-2 was expressed on the granulosa cells, however, Bax was not expressed. DNA fragmented cells were expressed on the inner granulosa cell layer of atretic follicles. CONCLUSION: Fas, Fas-ligand, and Bax may be responsible for the follicular atresia and Bcl-2 may be involved in the follicular survival in the human ovary.