Effects of polyphyllin Ⅵ on the proliferation and apoptosis of glioma cells and potential mechanism / 中国药房

OBJECTIVE To investigate the effects of polyphyllin Ⅵ（PPⅥ） on the proliferation and apoptosis of glioma cells and potential mechanism. METHODS Using human glioma LN229 cells as objects， MTT assay was used to detect the survival rate after treated with different concentrations of PPⅥ [0 （control group）， 1， 2， 4， 8， 16， 32， 64 μmol/L] for different time （24， 48， 72 h）. The clone formation experiments were adopted to detect the number of cell clones and clone formation rate after being treated with different concentrations of PPⅥ [0 （control group）， 2， 4， 8 μmol/L] for 14 days. The flow cytometry and Western blot assay were used to detect the apoptotic rate of cells， the expressions of apoptosis-related protein [B cell lymphoma-2 （Bcl-2）， Bcl-2 associated X protein （Bax）， cleaved caspase-3]， and the expressions of related proteins of Fas/Fas ligand （FasL） death receptor pathway and protein kinase B （Akt）/glycogen synthesis kinase-3β （GSK-3β） pathway after being treated with different concentrations of PPⅥ [0（control group）， 4， 8 μmol/L] for 24 h. RESULTS Compared with the control group， the survival rate of cells， the number of clones and clone formation rate， the protein expression of Bcl-2， and the phosphorylation levels of Akt and GSK-3β protein were decreased significantly in different concentration groups of PPⅥ （P＜0.05 or P＜0.01）. The apoptotic rate， the protein expressions of Bax， cleaved caspase-3， Fas， FasL and cleaved caspase-8 were increased significantly （P＜0.05 or P＜ 0.01）. CONCLUSIONS PPⅥ can inhibit the proliferation and induce the apoptosis of human glioma LN229 cells， which may be related to the activation of the Fas/FasL death receptor pathway and the inhibition of the Akt/GSK-3β pathway.

Effects of FasL inhibitors on corneal apoptosis and Treg after corneal transplantation in rats / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To explore the effect of intravitreal injection FasL inhibitors on corneal apoptosis, Fas, FasL expression, Treg numbers in blood and lymph nodes and rejection index in rats after corneal transplantation.METHODS:A total of 24 SD rats(24 eyes)who received penetrating keratoplasty were randomly divided into two groups: PBS group received intravitreal injection of PBS(12 rats, 12 eyes)and FasL inhibitor group(12 rats, 12 eyes). Rejection index was recorded every week and blood samples and lymph node were collected at 1, 3 and 5wk after surgery to analyze the proportions of Treg. Corneal tissue was collected for detecting the expression of Fas and FasL and number of apoptosis.RESULTS: The expression of Fas, FasL in FasL inhibitor group decreased significantly compared with the PBS group(all P<0.05); Corneal cell apoptosis significantly decreased in FasL inhibitor group, and it was the lowest at 5wk after surgery; Treg numbers in blood and lymph nodes significantly increased in FasL inhibitor group at 3wk after surgery(all P<0.05); rejection index of corneal transplantation in the FasL inhibitor group was significantly lower than that of PBS group(all P<0.05).CONCLUSION:Intravitreal injection of FasL inhibitors after corneal transplantation could reduce the apoptosis in all layers of cornea, increase the number of Tregs in blood and lymph nodes, and alleviate rejection.

Fas cell surface death receptor/Fas ligand genetic variants in gastric cancer patients: A case-control study

Background & objectives: Various studies have suggested a correlation between Fas cell surface death receptor/Fas ligand (FAS/FASL) variants and multiple types of cancers. The present study aimed to investigate the association between FAS-670A/G and FASL-844C/T and the synergistic effects of both variants on the risk of gastric cancer (GC) in the Kurdish population of west of Iran. Methods: This study was conducted by polymerase chain reaction-restriction fragment length polymorphism technique using MvaI and BsrDI restriction enzymes in 98 GC patients and 103 healthy control individuals. Results: According to the obtained results, a significant association (P=0.008) of FASL polymorphism among GC patients and the control group was detected. Furthermore, no significant differences were found in the FAS polymorphism frequencies between GC patients and the control group. Codominant and dominant models in FASL polymorphism showed significant protective effects against GC [odds ratio (OR)=0.307, 95% confidence interval (CI) (0.134-0.705), P=0.005; OR=0.205, 95% CI (0.058-0.718), P=0.013 and OR=0.295, 95% CI (0.129-0.673), P=0.004 for models of codominant CC vs. CT, codominant CC vs. TT and dominant, respectively]. Furthermore, the presence of both FAS-670G and FASL-844T alleles represented a significant protective effect against GC occurrence [OR=0.420, 95% CI (0.181-0.975), P=0.043]. Interpretation & conclusions: So far, we believe this is the first study, the results of which suggest that FASL gene variation and its synergistic effects with FAS gene could be associated with the risk of GC in the Kurdish population in the west of Iran

Transplantation of human stem cells from the apical papilla for treating dextran sulfate sodium-induced experimental colitis / 中国组织工程研究

BACKGROUND: Stem cells from the apical papilla (SCAP) play important roles in the formation and development of dental roots. However, the immune-modulating capacity of SCAP has not been fully elucidated. OBJECTIVE: To test the therapeutic effects of transplantation of SCAP on dextran sulfate sodium-induced experimental colitis. METHODS: Twenty-four C57/BL6 mice were equally divided into four groups (normal control, positive control, SCAP treatment group, and FasL-knockdown SCAP group), and latter three groups of mice were induced to acute experimental colitis by 3% dextran sulfate sodium in drinking water. At day 3 after modeling, model mice were treated with PBS, human SCAP (2×106 cells), and FasL-knockdown SCAP via intraperitoneal injection, respectively. Inflammation was evaluated by measuring body mass and length of the colon, detecting levels of interleukin 1β, interleukin 6 and tumor necrosis factor α, as well as histological analyses at day 10 after modeling. Levels of Tregs in mesenteric lymph nodes in mice were detected using flow cytometric analysis. RESULTS AND CONCLUSION: SCAP transplantation could ameliorate the inflammation in dextran sulfate sodium-induced colitis mice, and body mass loss and symptoms were significantly improved. Pathological score and the levels of three inflammatory cytokines in the colon tissue decreased significantly. Flow cytometric analysis revealed an increased level of Tregs in mesenteric lymph nodes. Knocking down of FasL gene in SCAP abrogated the therapeutic effects of SCAP in ameliorating dextran sulphate sodium-induced colitis. Therefore, Fas-FasL pathway played an important role in the underlying mechanism of the immune-modulating capacity of SCAP.

Variation of fas/fasl pathway in limb ischemia-reperfusion lung injury rats after etomidate post-treatment / 中国组织工程研究

BACKGROUND: To date, it has been confirmed that etomidate pre-treatment can reduce the damage of remote organs caused by limb ischemia-reperfusion, but whether etomidate post-treatment has protective effect on remote organs and its mechanism has been rarely reported. OBJECTIVE: To investigate the influence of etomidate post-treatment on limb ischemia-reperfusion lung injury. METHODS: A rat model of limb ischemia-reperfusion lung injury was prepared by clamping the bilateral femoral arteries for 2 hours and reperfusion for 3 hours. After 2 hours of limb ischemia, I/R group experienced the process of limb ischemia-reperfusion; I/R+ETO group, I/R+Dex 0.2 group, I/R+Dex 0.5 group and I/R+Dex 1.0 group, besides the model of limb ischemia-reperfusion, were injected with etomidate 1.0 mg/kg and dexamethasone 0.2, 0.5 and 1.0 mg/kg respectively through tail vein. At 3 hours of reperfusion, blood samples were extracted from the carotid artery, blood gas analysis was performed and the partial pressure of blood oxygen (PaO2) was recorded. The pathological changes were detected by immunohistochemistry. Apoptotic index was detected by Hoechst 33258 staining and wet/dry weight ratio was detected. Fas protein and Fasl mRNA of lung tissue were detected by western blot and RT-PCR respectively. Tumor necrosis factor-α and interleukin-1β levels were detected by ELISA. RESULTS AND CONCLUSION: Compared with the I/R group, PaO2 increased (P 0.05). To conclude, etomidate post-treatment can reduce lung injury caused by limb ischemia-reperfusion in rats, and its mechanism may be related to the down-regulation of Fas/FasL. In the statistical point of view, etomidate 1.0 mg/kg has the potency intensity of reducing lung injury, almost equivalent to dexamethasone 0.5 mg/kg.

Effect of electroacupuncture at different stages on the expression of Fas and FasL in brain tissue of rats with traumatic brain injury / 针刺研究

OBJECTIVE: To investigate the expression of apoptosis-related proteins Fas and FasL in the brain tissue of rats with traumatic brain injury and the effect of electroacupuncture on the expression of Fas and FasL, so as to explore the effective time window of electroacupuncture in the treatment of traumatic brain injury. METHODS: Sprague-Dawley rats were randomly divided into blank group, sham-operation group, model group, and electroacupuncture treatment groups 1, 2, and 3. Traumatic brain injury was induced by the modified Feeney free-fall impact device, and for the rats in the electroacupuncture treatment groups 1, 2, and 3, electroacupuncture started at 4 hours and on days 3 and 7, respectively, after modeling and lasted to day 14. The Morris water maze test was used to evaluate learning and memory ability, and immunofluorescence assay and Western blot were used to observe the changes in the expression of Fas and FasL in traumatic brain tissue. RESULTS: Compared with the blank group and the sham-operation group, the model group had a lower percentage of time spent in the target quadrant from the 3rd day folowing modeling; after electroacupuncture intervention, the electroacupuncture treatment groups showed a gradual increase in the time spent in the target quadrant, and on day 7,10 and 14, electroacupuncture treatment group 1 had a significantly higher percentage than the model group (P<0.05). On day 14, electroacupuncture treatment group 2 had a significantly higher percentage than the model group (P<0.05). After electroacupuncture intervention, all groups except the blank group and the sham-operation group had increases in the expression of Fas and FasL in brain tissue, which reached the highest level on day 7 after modeling and then tended to decrease; compared with electroacupuncture treatment groups 2 and 3 and the model group, electroacupuncture treatment group 1 had significant reductions in the expression of Fas and FasL (P<0.05, P<0.01); compared with electroacupuncture treatment group 3 and the model group, electroacupuncture treatment group 2 had significant decreases in the expression of Fas and FasL (P<0.05) on day 14 after modeling; compared with the model group, electroacupuncture treatment group 3 had significant reductions in the expression of Fas and FasL in brain tissue on day 14 after modeling (P<0.05). CONCLUSION: Early electroacupuncture intervention can regulate the apoptosis receptor pathway by down-regulating Fas and FasL to exert a therapeutic effect on traumatic brain injury and help with the recovery of cognition and memory ability after traumatic brain injury.

Experimental study on the effect of hedysarum polybotys saccharides and selenizated hedysarum polybotys saccharides on oral squamous cancer cells in vitro / 口腔疾病防治

Objective@#To study the effects of hedysarum polybotys saccharides (HPS) and selenizated hedysarum polybotys saccharides (SE-HPS) on the oral squamous cancer cell line SCC25.@*Methods@#Different concentrations (0, 10, 25, 50, 100, 200, 400 μg/ml) of HPS and SE-HPS were added to SCC25 cells in the logarithmic growth stage. Cell proliferation was detected by the CCK-8 method, apoptosis was detected by flow cytometry, and apoptosis-related indexes were observed by RT-qPCR and Western blotting.@*Results @#The concentrations of HPS and SE-HPS inhibited the proliferation of SCC25 cells. The inhibitory effect of 50 μg/mL HPS and SE-HPS on the proliferation of SCC25 cells was the strongest and was time-dependent. The inhibition effect significantly increased within 48 h, and the effect was achieved after 48 h. At the plateau stage, SE-HPS inhibited the proliferation of SCC25 cells more strongly than HPS (P < 0.05). The results of flow cytometry showed that 50 μg/mL HPS and SE-HPS acted on SCC25 cells for 48 h, and the apoptotic rates were 25.8% and 30.8% respectively. Compared with the control group (0 μg/mL HPS and SE-HPS), the difference was statistically significant (P < 0.05). RT-qPCR and Western blotting showed that 50 μg/mL HPS and SE-HPS acted on SCC25 cells for 48 h, and the mRNA and protein expression levels of the apoptosis gene Fas/FasL were upregulated. The difference was statistically significant (P < 0.05).@*Conclusion@# Both HPS and SE-HPS can inhibit the proliferation of SCC25 oral cancer cells, but SE-HPS is superior to HPS and can induce apoptosis through the Fas/Fasl pathway.

Preparation of mouse anti-human B7-2 monoclonal antibody and analysis of its role in inducing Fas and FasL on the surface of 8266 cells / 中华微生物学和免疫学杂志

Objective To prepare a mouse anti-human B7-2 monoclonal antibody ( McAb) and to study its effect on the induction of death-related molecules on the surface of tumor cells. -ethods Trans-genic cells, L929-B7-2, were used as the immunogen to immunize BALB/c mice. Through cell fusion, mul-tiple screening by immunofluorescence labeling and continuous subcloning, the hybridoma secreting B7-2 McAb was obtained. Biological characteristics of the McAb were analyzed using Ig subclass identification test strip, antigenic site competition inhibition assay and specific cell membrane molecules binding test. McAb was prepared through inducing ascites in vivo and then purified by protein G affinity chromatography. The purified McAb was co-cultured with 8266 cells, naturally expressing B7-2 molecules, to observe the expres-sion of Fas and FasL on cell surface by flow cytometry ( FCM) . Results The prepared B7-2 McAb labeled as 12G4 was successfully obtained with a titer of 0. 1 μg/5×105 cells. Its heavy and light chains were IgG2b and κ, respectively. The concentration of the purified ascites-derived antibody was 1. 61 mg/ml. FCM re-sults showed that the 12G4 McAb recognized cell membrane molecules well with a positive binding rate of 89. 6％ to 8266 cells. The mean value of the Fas molecule on the cell surface increased after incubating with 20 μg/ml of 12G4 McAb for 12 h and reached the peak of 62575. 8 at 48 h, which was significantly higher than the maximum value of 57135. 4 in the IgG control group (P<0. 05). After culturing the cells with 20μg/ml of 12G4 McAb for 12 h, the expression of FasL on the cell surface also increased and reached the maximum of 7. 98％ at 48 h, which was significantly higher than the 1. 10％ in the IgG control group ( P<0. 05). Conclusions B7-2 McAb was successfully prepared. It could be used to induce the expression of some death-related molecules on the surface of tumor cells.

Effect of Tripteryginum wilfordii polyglycoside on Fas/FasL and p38 MAPK signaling pathway expression in ulcerative colitis rats / 中国药理学通报

Aim: To investigate the expression changes of Fas/FasL and its related signaling pathway p38 MAPK in ulcerative colitis(UC) rats and the effects of Tripterygium wilfordii polycoride (TWP). Methods: TNBS and ethanol enema method were adopted to build the UC rat model, and colon inflammation in rats was assessed; the differential expression of Fas/FasL was analyzed by expression profiling method, and verified by real-time PCR. Subsequently, pathway analysis in DAVID database showed that Fas/FasL might involve p38 MAPK signaling pathway in regulating UC inflammatory activities, and the related molecules in this signaling pathway were analyzed by the expression profiling method and verified by real-time PCR. Finally, the expression of the terminal inflammatory factors of the pathway were detected by RT-PCR and Western blot. Results: In UC rats, the gross morphological damage and histopathological injury scores significantly increased, which could be reduced by TWP. The trend of FasL was in consistent with that of MAPK14 (p38α), TNF-α and IL-1β in Fas/FasL system and the p38 MAPK signaling pathway, and the expression of them were up-regulated in inflammatory activity, which can be down-regulated by TWP. Conclusion: TWP can regulate the inflammatory activity of UC through Fas/FasL system and the p38 MAPK signaling pathway.

Effect of 3 neuroprotective measures on expressions of Fas/FasL in brain tissue of neonatal rats with hypoxicischemic brain damage / 中国中西医结合急救杂志

Objective To observe the effects of 3 neuroprotective measures on the expressions of apoptosis-related factors and their ligands (Fas and FasL) in brain tissue of neonatal rats with hypoxic ischemic brain injury. Methods One hundred and twenty Wistar rats 7 days old were selected as experimental subjects, the rats were divided into four groups: neural stem cell, erythropoietin (EPO), ω-3 unsaturated fatty acid treatment groups and hypoxic ischemic brain damage model group according to random number table method, with 30 rats in each group. Neural stem cell group, EPO group and ω-3 unsaturated fatty acid group were respectively injected with neural stem cells, EPO and ω-3 unsaturated fatty acid, each 5 mL via tail vein after modeling; the hypoxic ischemic brain damage model group was given equal volume of normal saline. At 6, 12, 24, 48 and 72 hours after administration of drug, 6 rats were sacrificed in each group, brain tissue was taken, the mRNA expression levels of Fas/FasL, protein expression levels of Toll-like receptor 4 (TLR4), nuclear transcription factor-κB (NF-κB), tumor necrosis factor-α (TNF-α), interleukin (IL-1β, IL-6) and cell apoptotic rate in hippocampus tissue were measured. Results ① mRNA expressions: the mRNA expressions of Fas and FasL of the 3 experimental groups were significantly lower than those of the hypoxic ischemic brain damage model group, the degrees of descent after administration for 24 hours were the most significant, neural stem cell treatment group < EPO treatment group < ω-3 unsaturated fatty acid treatment group < hypoxic ischemic brain damage model group [Fas mRNA expression (2-ΔΔCt): 140.5±2.9, 156.4±2.5, 165.2±2.7 vs. 173.7±2.8, FasL mRNA expression (2-ΔΔCt): 143.1±4.3, 154.6±1.5, 160.7±1.4 vs. 174.7±2.8], the differences were statistically significant (all P < 0.05). ② Protein expressions: the protein expressions of TLR4, NF-κB, TNF-α, IL-1β, IL-6 of the 3 experimental groups were significantly lower than those of the hypoxic ischemic brain damage model group (TLR4/GAPDH: 0.7±0.2, 0.6±0.1, 0.2±0.1 vs. 1.4±0.1; NF-κB/GAPDH: 6.7±0.4, 5.3±0.1, 1.1±0.2 vs. 11.2±0.3; TNF-α/GAPDH: 14.3±1.4, 11.2±1.2, 3.2±2.1 vs. 23.2±0.5; IL-1β/GAPDH: 9.4±0.2, 7.4±0.3, 2.2±0.3 vs. 13.4±0.1; IL-6/GAPDH: 36.2±4.4, 39.3±1.5, 26.2±2.1 vs. 51.4±1.4, all P < 0.05), the protein expression levels of above indexes in neural stem cell treatment group < those of EPO treatment group < those of ω-3 unsaturated fatty acid treatment group < those of hypoxic ischemic brain damage model group. ③ Apoptotic rates:after drug administration, the apoptotic rates of the ω-3 unsaturated fatty acid group, EPO treatment group, neural stem cell treatment group were obviously lower than the rate of model group [(3.7±0.3)%, (3.4±0.2)%, (2.5±0.1)% vs. (5.5±0.4)%, all P < 0.05]. Conclusion The mRNA expressions of Fas/FasL in the brain of neonatal rats with hypoxic-ischemic brain damage are high, and the treatment with each of the following agents; neural stem cells, EPO and ω-3 unsaturated fatty acid can reduce the mRNA expressions of Fas/FasL in such rats' brain tissues.

Expression of Fas/FasL in PC12 cell apoptosis induced by ropivacaine / 临床麻醉学杂志

Objective To detect the expression of Fas and FasL in ropivacaine-induced rat phechromocytoma (PC12)cells apoptosis and the mechanism of its neurotoxicity.Methods PC12 cells were treated with different concentrations of ropivacaine (0.1,0.5,1,2,and 4 mmol/L)for 24 h to establish a cellular neurotoxicity model,and the cell viability were assessed by CCK-8.The cells were finally divided into 3 groups randomly:0.5 mmol/L group,2 mmol/L group and control group. After the cells were cultured for 24 h,morphological changes of cells were observed under optical mi-croscope (add the 1 mmol/L group),apoptosis was assessed by flow cytometer,the expression of Fas and FasL were assessed by immunofluorescence.Results Compared with the control group,the cell viability of 0.5 mmol/L and 2 mmol/L group decreased significantly (P<0.05),the cells exhibited obvious morphologic abnormalities (including the 1 mmol/L group),the apoptotic rate increased sig-nificantly (P<0.05),the expression of Fas and FasL increased significantly (P<0.05);Compared with 0.5 mmol/L group,the apoptotic rate and expression of Fas,FasL of 2 mmol/L group increased significantly (P<0.05).Conclusion Ropivacaine explosure induces apoptosis in PC12 cells,which might be related with the up-regulation of Fas/FasL.

Xanthotoxin induces apoptosis in SGC-7901 cells through death receptor pathway / 中草药·英文版

Objective: To investigate the effects of xanthotoxin from Apiaceae medicinal plants on cell proliferation and apoptosis, and explore its mechanism of action against human gastric carcinoma SGC-7901 cells in vitro. Methods: SGC-7901, HepG-2, MCF-7, and A549 cells were treated with different concentrations of xanthotoxin (10, 20, 60, 80, 100, 120, 140, and 160 µg/mL) for 48 h, and the cell viability (IC50) was determined by MTT assay; Xanthotoxin-induced apoptosis in cells was observed by using Hoechst 33258 Staining Kit and Annexin V-FITC Apoptosis Detection Kit; Flow cytometry was used to detect apoptosis related proteins of Fas/FasL, Bid, and DR5/TRAIL proteins in human gastric carcinoma SGC-7901 cells after being treated by xanthotoxin; The influence of xanthotoxin on Caspase-8 protein expression in the cells was determined by Flouormetric Assay Kit. Results: Xanthotoxin obviously inhibited SGC-7901, HepG-2, MCF-7, and A549 cells proliferation, and its inhibition was in a concentration-dependent manner; flow cytometry results showed that in a certain concentration range, xanthotoxin can increase the expression levels of Fas/FasL and DR5/TRAIL proteins in a concentration-dependence manner. The content of Bid protein in cells was increased, and it showed concentration-dependence. Conclusion: Xanthotoxin may induce SGC-7901 cells apoptosis in a certain concentration range through the Fas/FasL protein mediated death receptor pathway, or by DR5/TRAIL mediated death receptor pathway, and increase the expression level of death receptor protein, activation Caspase-8, activating downstream effect factor, inducing cell apoptosis, or activate Caspase-8 cutting activate protein Bid, and then enter the mitochondrial pathway, induction of apoptosis.

Effects of Electro-acupuncture at “Weizhong” (BL40) on Expressions of Fas and FasL in Rats with Lumbar Disc Degeneration / 中国中医药信息杂志

Objective To observe the effects of electro-acupuncture (EA) at “Weizhong” (BL40) on expressions of Fas and FasL in rat models with lumbar disc degeneration; To discuss its mechanism of action.MethodsThirty SD rats were randomly divided into sham-operationgroup, model group and EA group, with 10 rats in each group. The sham-operation group was treated with sham operation to incide local skin; the model and EA groups established rat model of lumbar disc degeneration by puncturing the annulus fibrosus. Four weeks after modeling, rats in the sham-operation and model groups received fixed treatment under identical condition, and rats in the EA group were treated with EA at “Weizhong” (BL40) in 20 minutes, once a day, for 4 weeks. After treatment, Western blot and RT-PCR technology were used to test the protein and mRNA expression levels of Fas and FasL.ResultsThe expressions of Fas and FasL in the model group were higher than those in the sham-operation group (P<0.05); protein and mRNA expression levels of Fas and FasL in the EA group were lower than those in the model group (P<0.05). ConclusionEA at “Weizhong” (BL40) can reduce the expressions of Fas and FasL to inhibit the development of lumbar disc degeneration.

Clinical research about the correlation between Fas、FasL protein and breast cancer / 中国现代普通外科进展

Objective:To investigate the clinical significance of the expression of Fas and FasL in breast cancer tissue and their relationship with the prognosis of breast cancer.Methods:Immunohistochemical technique was used to detect protein expression proportion and the level of Fas and FasL in 60 cases of breast cancer and 10 cases of benign breast tumor tissues.the experimental data were analyzed using semi-quantitative count and SPSS22.Results:The positive rate of Fas,FasL in breast cancer and benign breast tumor tissues were 78.33％ (47/60),80％ (8/10) and 95％ (57/60),70％(7/10),the positive rate of Fas protein in breast cancer tissues was higher than that of benign breast tumors,the positive rate of FasL protein in breast cancer was lower than that in benign breast tumor tissue.Fas、FasL protein expression were no significantly correlated with that of breast cancer staging,age,molecular typing,HER-2 and axillary lymph node status.but The strong expression of Fas,FasL protein were significantly correlated with that of the axillary lymph node status and the expression of ki-67.Conclusion:The strong expression of Fas,FasL protein may be associated with breast cancer metastasis and progress,and they could be as indicators of the evaluation of prognosis of breast cancer.

Study on role of Fas/FasL in pathogenesis of primary immune thrombocytopenia / 重庆医学

Objective To explore the role of Fas and FasL system in the pathogenesis of adult primary immune thrombocyto‐penic purpura (ITP) .Methods The peripheral anticoagulant venous blood samples were collected from the patients with newly di‐agnosed ITP and healthy adults .The expression rates of Fas and FasL in Th/Tc ,Th1/Th2 ,Tc1/Tc2 cells and their expression rates at platelet surface were detected by flow cytometry .Results The Fas and FasL expression rates on the surface of Th ,Th1 ,Th2 , Tc ,Tc1 and Tc2 in the ITP group were increased compared with the healthy control group(P0 .05) .Conclusion The Fas and FasL expression on T cell subsets and platelet surface in ITP patients is abnormal , which may be related with the pathogenesis of ITP .

A study on the relationship between Fas-670 G>A and Fasl-844 T>C polymorphisms and the susceptibility to lung cancer / 中国癌症杂志

Background and purpose:Lung cancer is one of the leading causes of cancer death in the world. The aim of this study was to evaluate whether Fas-670 G>A and Fasl-844 T>C polymorphisms were associated with the risk of lung cancer.Methods:Data from 400 lung cancer patients with speciifc histological diagnosis were collected from 2010 to 2015. Meanwhile, data from matched healthy controls with the same gender and ±5 years were also collected. The genotypes of Fas-670 G>A and Fasl-844 T>C polymorphisms were determined by TaqMan lfuorescent probe method, and the results were analyzed using SPSS 16.0 software.Results:A total number of 386 cases and 394 controls were successfully genotyped. Compared with AA genotypeofFasgene, the GA and GG genotype carriers had no signiifcantly increased risk of lung cancer.The OR values were 1.05 (95%CI: 0.77-1.44) and 0.77 (95%CI: 0.81-1.99) respectively. Compared with TT genotype ofFasl gene, the CT and CC genotype carriers had signiifcantly increased risk of lung cancer. The OR values were 1.37 (95%CI: 1.01-1.86) and 1.74 (95%CI: 1.09-2.77), respectively. Conclusion:Fasl-844 T>C polymorphism may be involved in lung cancer risk but not Fas-670 G>A polymorphism.

Expression of mir-199a in ulcerative colitis rats induced by 2, 4, 6-trinitrobenzene sulfonic acid/ethanol and study on the effect of tripteryginum wilfordii polyglycoside / 中国药学杂志

OBJECTIVE: To explore the expression of miR-199a in ulcerative colitis (UC) rats induced by 2, 4, 6-trinitrobenzene sulfonic (TNBS)/ethanol and the study on the effect of TWP on them. METHODS: Through injecting TNBS/ethyl alcohol acid liquid into the anus of the rats to establish the UC rat model. The colitics commom morphous damage and grade the histopathological score (CMDI) of colon mucosa injury were evaluated. Chip analysis and Real-time PCR were used to verify the expression of miR-199a in each colon mucosa tissue. Based on the expression profile, the downstream target genes mRNA in milwalk database was selected, then the expression of target genes mRNA by Real-time PCR in each group was veritied, at last the relevant signal pathway in the DAVID database was analyzed. Doing these to analyse the target gene mRNA regulated by the miR-199a in the inflammatory activity of UC. RESULTS: Compared with the model group, TWP high dose group was significantly lower on gross morphological damage score and histopathological injury score(P < 0.01). Chip analysis showed that in model group, the expression of miR-199a was significantly higher than the normal group(P < 0.01), and expression of the AZA group was significantly lower than the model group(P < 0.01, P < 0.05). The expression of miR-199a-3p in medium dose group and the expression of miR-199a-5p in high dose group were significantly lower than the model group(P < 0.05). The results of Real-time PCR showed that expression of miR-199a in the model group was significantly increased than that in the normal group(P < 0.01). The expression of miR-199a-3p in TWP medium dose group, high dose group and AZA group were decreased than that in model group(P < 0.05). Meanwhile, the expression of miR-199a-5p in TWP medium dose group was decreased than that in model group(P < 0.05). The gene expression profile showed that FASL was the target gene of miR-199a. In the model group, the expression of FASL was higher than that in the normal group. The expression of FASL in AZA group was significantly decreased than that in the model group(P < 0.01). The results by the Real-time PCR of the target gene FASL showed that in the model group, the expression of FASL was higher than that in the normal group (P < 0.01). The expression of FASL in medium dose group, high dose group and AZA group were significantly decreased than that in the model group (P < 0.01). CONCLUSION: miR-199a is up-regulated in TNBS/Ethanol UC rats, and FASL is the downstream target gene of miR-199a. TWP can reduce the UC's inflammatory effectively and decrease the up-regulated miR-199a in UC. FASL is up-regulated in UC's inflammatory activity. TWP can reduce downstream target gene FASL of miR-199a.

Icariin induces apoptosis of esophageal cancer cell through enhancing Fas-FasL expression activity / 重庆医学

Objective To investigate the inhibitory effect of icariin(ICA) on the xenograft tumors growth of esophageal car‐cinoma and to preliminarily investigate its mechanism .Methods The MTT assay and Giemsa staining were applied to detect and observe the in vitro inhibitory effect of ICA on esophagus cancer cell lines Eca‐109 and TE‐13 .The xenograft tumor model of nude mouse esophagus cancer cell was constructed and divided into 3 groups ,6 cases in each group .Each mouse in the experimental group was intraperitoneally injected by ICA 50 mg/kg ;while the control group was injected by the same volume of normal saline and the positive control group was injected by cis‐platinum 2 mg/kg ,once every 2 days ,a total of 14 days .The tumor volume was measured once per 3 d .After experiment ,the tumor weight was measured;the TUNEL staining was used to observe the morphological chan‐ges and cell apoptosis of tumor tissue in each group .The changes of Fas and FasL protein expression in tumor tissues were analyzed by immunohistochemistry .The FasL and IFN‐γlevels in peripheral blood were tested by the ELISA assay .Results ICA exerted no obvious inhibitory effect on the proliferation of Eca‐109 and TE‐13 cell in vitro .The average volume and weight of xenografts tumor had statistical difference between the experimental group and the positive control group (P<0 .05) .The TUNEL staining results showed that the tumor tissues had obvious apoptosis ,the number of apoptosis cells was significantly increased compared with the control group(P<0 .05) .The immunohistochemistry experimental results showed that the expression of Fas and FasL was signifi‐cantly increased(P<0 .05) .The ELISA experimental results demonstrated that the FasL and IFN‐γlevels of peripheral blood in the experimental group were significantly increased(P<0 .05) .Conclusion ICA had no obvious inhibitory effect on esophageal cancer cell proliferation in vitro ,but could induce in vivo apoptosis through the Fas expression and secretion of FasL and IFN‐γ,thus plays the role of anti‐esophageal cancer .

Low concentrations of doxycycline attenuates FasL-induced apoptosis in HeLa cells

BACKGROUND: Doxycycline (DC) has been shown to possess non-antibiotic properties including Fas/Fas Ligand (FasL)-mediated apoptosis against several tumor types in the concentration range of 10-40 µg/mL. However, the effect of DC in apoptotic signaling at much low concentrations was not studied. METHODS: The present study investigated the attenuation effect of low dose of DC on FasL-induced apoptosis in HeLa cell by the methods of MTT assay, fluorescence microscopy, DNA fragmentation, flow cytometry analysis, and western blotting. RESULTS AND CONCLUSION: In the present findings we showed that low concentration of DC (<2.0 µg/mL) exhibited protective effects against FasL-induced apoptosis in HeLa cells. FasL treatment to HeLa cells resulted in a concentration-dependent induction of cell death, and treatment with low concentrations of DC (0.1-2 µg/mL) significantly (p < 0.001) attenuated the FasL-induced cell death as measured by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazo-lium bromide (MTT) assay. Further, the FasL-induced apoptotic features in HeLa cells, such as morphological changes, DNA fragmentation and cell cycle arrest was also inhibited by DC (0.5 µg/mL). Tetracycline and minocycline also showed similar anti-apoptotic effects but were not significant when compared to DC, tested at same concentrations. Further, DC (0.01-16 µg/mL) did not influence the hydrogen peroxide- or cisplatin-induced intrinsic apoptotic pathway in HeLa cells. Protein analysis using Western blotting confirmed that FasL-induced cleavage/activation of cas-pase-8 and caspase-3, were inhibited by DC treatment at low concentration (0.5 µg/mL). Considering the overall data, we report for the first time that DC exhibited anti-apoptotic effects at low concentrations in HeLa cells by inhibition of caspase activation via FasL-induced extrinsic pathway.

Roles and mechanisms of Fas/FasL in the apoptosis of HL-60 cells induced by euphorbiasteroid / 国际肿瘤学杂志

Objective To investigate the effect of euphorbiasteroid on inducing the apoptosis of HL-60 cells and demonstrate whether the Fas/FasL signaling pathway is involved in the induction of apoptosis. Methods HL-60 cells were treated with dose of 2.5,10,40 μg/ml of euphorbiasteroid in vitro for 24 h respectively.After that,cell counting Kit-8 was used to detect cell proliferation.The morphology of HL-60 cells were observed under light and fluorescent microscopy.The early cell apoptosis was detected by using flow cytometry with Annexin Ⅴ-FITC /PI double staining.The expressions of Fas,FasL,caspase-8 and caspase-3 mRNA were analyzed by the method of RT-PCR.The activities of caspase-8 and caspase-3 were examined by chromatometry.Results Compared with 1640 control group,HL-60 cell proliferation was inhibited significant-ly by euphorbiasteroid.The inhibition rates were (34.9 ±3.7)%,(54.6 ±5.2)% and (61.3 ±4.3)%respectively.Moreover,HL-60 cells exhibited typical morphological features.Early cell apoptosis rates of HL-60 cells were (23.4 ±3.1)%,(35.7 ±4.3)% and (53.2 ±3.9)% respectively.Furthermore,the expressions of Fas,FasL,caspase-3 and caspase-8 mRNA were up-regulated significantly after euphorbiasteroid administration in a dose-dependent manner (P<0.01 ).After treated with euphorbiasteroid,the activities of caspase-8 and caspase-3 were significantly enhanced (P<0.01 ).Conclusion The up-regulation effect of euphorbiasteroid on Fas/FasL signaling pathway might contribute to the apoptosis of HL-60 cells.