FasL Protein Expression Research Articles

Long non-coding RNA XIST modulates cisplatin resistance by altering PDCD4 and Fas-Lexpressions in human nasopharyngeal carcinoma HNE1 cells in vitro

To explore the role of long non-coding RNA (lncRNA) X inactive specific transcript (XIST) in modulating cisplatin (DDP) resistance of human nasopharyngeal carcinoma cells and investigate the possible mechanism. Realtime PCR was performed to detect the expression of XIST in cisplatin-resistant human nasopharyngeal carcinoma cell line HNE1/DDP. The effects of up-regulation and down-regulation of XIST on DDP resistance, proliferation and apoptosis of HNE1/ DDP cells were assessed using MTT assay, EdU assay and flow cytometry. Western blotting was used to detect the changes in the expressions of programmed cell death 4 (PDCD4) and Fas ligand (Fas-L) proteins in the cells in response to up-regulation or down-regulation of XIST. The expression of XIST was significantly up-regulated in HNE1/DDP cells in comparison with HNE1 cells (0.57±0.06 vs 0.1±0.02, P < 0.05). Down-regulation of XIST significantly decreased while up-regulation of XIST obviously increased DDP resistance of HNE1/DDP cells (P < 0.05). Down-regulation of XIST significantly reduced the proliferation (6.17 ± 1.93 vs 16.59 ± 4.86, P < 0.05) and enhanced apoptosis [(18.04 ± 4.72)% vs (4.22 ± 1.65)%, P < 0.05], while upregulating XIST enhanced the proliferation (25.40±7.21 vs 13.16±3.95, P < 0.05) and inhibited apoptosis [(2.82±0.88)% vs (6.46± 1.75)%, P < 0.05] in HNE1/DDP cells. Down-regulation of XIST significantly increased the protein expressions of PDCD4 and Fas-L (P < 0.05) in HNE1/DDP cells, and up-regulation of XIST resulted in reverse changes in PDCD4 and Fas-L expressions (P < 0.05). XIST is up-regulated in HNE1/DDP cells, and down-regulation and up-regulation of XIST expression reduce and increase DDP resistance of the cells, respectively, possibly as a result of changes in the expressions of PDCD4 and Fas-L.

Baicalin protects mouse testis from injury induced by heat stress.

Heat stress has been documented to reduce reproductive performance of female animals through injury to germ cells, with few studies available in male animals. The objectives of this study were to evaluate protective effects of baicalin on testicular tissue damage of mice subjected to heat stress and its related mechanisms. In this experiment, A total of forty mice were divided into four groups, including control group (C), baicalin group (B), heat stressed group (H) and heat stress with baicalin treatment (H+B) group. Morphological changes, activities of antioxidant enzymes and apoptosis-related parameters in the mice testes tissue were monitored. The results showed that the process of spermatogenesis in mice testis was impaired and the cellular apoptosis increased due to acute heat stress at 41°C. Interestingly, the tissue damage was alleviated with the significant (P < 0.05) increase in the activities of SOD, CAT and GSH-Px enzymes, decrease (P<0.05) in MDA content and number of cellular apoptosis recorded in mice of H+B group compared with those in mice from H group. In addition, the Fas, FasL and P-JNK protein expressions were significantly (P<0.05) increased; and apaf-1, caspase-3, -9 were slightly expressed in the H group, while there was no difference in Bcl-2 expression, compared with C, B and H+B groups. The above results clearly indicate that heat stress induces macroscopic/apoptotic and oxidative changes in the testicular tissue of mice; these changes are alleviated by Baicalin through increasing anti-oxidative enzyme activities and possibly through blocking Fas/FasL pathway.

Resveratrol analog, N-(4-methoxyphenyl)-3,5-dimethoxybenzamide induces G2/M phase cell cycle arrest and apoptosis in HeLa human cervical cancer cells

In this study, several resveratrol analogs were synthesized and evaluated in search of a more effective anti-proliferative resveratrol analog. Among the evaluated resveratrol analogs, we have identified N-(4-methoxyphenyl)-3,5-dimethoxybenamide (MPDB) as a potent anti-proliferative compound. Treatment with MPDB resulted in G2/M phase cell cycle arrest, which was accompanied by alteration of G2/M-related protein expression and phosphorylation. MPDB-induced G2/M arrest was blocked by transfection of ATM/ATR siRNAs, indicating the critical role of ATM/ATR in G2/M phase arrest. In addition, treatment with MPDB displayed the activation of caspase and decreased Bcl-xl protein expression after 20 h in HeLa cells. Moreover, MPDB increased cytosolic cytochrome c release and Fas and Fas-L protein expression, indicating intrinsic and extrinsic apoptosis pathway, respectively. These results suggest that MPDB is a new and potent compound that induces ATM/ATR-dependent G2/M phase cell cycle arrest and apoptosis, implicating it as a putative candidate in the investment of cervical cancer therapy.

Regulatory effects of miRNA‑181a on FasL expression in bone marrow mesenchymal stem cells and its effect on CD4+T lymphocyte apoptosis in estrogen deficiency‑induced osteoporosis.

Post-menopausal osteoporosis is a bone formation disorder induced by estrogen deficiency. Estrogen deficiency facilitates the differentiation and maturation of osteoclasts by activating T lymphocytes. In our previous study, it was demonstrated that estrogen promotes bone marrow mesenchymal stem cell (BMMSC)-induced osteoclast apoptosis through downregulation of microRNA (miR)-181a and subsequent Fas ligand (FasL) protein accumulation. In the present study, the regulatory effects of miR-181a on FasL expression in BMMSCs and the apoptotic effects of BMMSCs on cluster of differentiation (CD)4+T lymphocytes were investigated. An ovariectomized mouse model of osteoporosis (OVX) was established and CD4+T lymphocytes were isolated from the bones of these mice. The results demonstrated that the number of CD4+T lymphocytes was increased in the OVX group compared within the control group, thus suggesting that estrogen deficiency may increase CD4+T lymphocyte number. CD4+T lymphocytes were subsequently co-cultured with estrogen-treated BMMSCs, after which it was demonstrated that estrogen significantly promoted the apoptosis of CD4+T lymphocytes. Western blot analysis indicated that estrogen promoted the apoptosis of CD4+T lymphocytes through regulation of FasL expression in BMMSCs in a concentration-dependent manner. Finally, miR-181a was transfected into BMMSCs, which were co-cultured with CD4+T lymphocytes in vitro and in vivo. The results revealed that miR-181a exerted a negative regulatory effect on BMMSC-induced CD4+T lymphocyte apoptosis by regulating FasL protein expression in BMMSCs; this maybe a key mechanism underlying the development of estrogen deficiency-induced osteoporosis.

Effects of Moxibustion on Ventricular Mass Index and Expression of Apoptosis Related Proteins in Myocardium of Rats with Chronic Heart Failure

To observe the effect of moxibustion on cardiac function and the expression of B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X (Bax), Fas, Fas ligand (FasL) in cardiomyocytes of chronic heart failure (CHF) rats, so as to explore its underlying mechanisms in preventing and treating CHF. SD rats were randomly divided into normal control, model, moxibustion, Captopril and moxibustion ＋ Captopril (M＋C) groups (n＝12 rats/group). The CHF model was established by intraperitoneal injection of Adriamycin (ADR, from 1 to 4 mg/kg, once every other day for 15 days). Mild moxibustion was applied to bilateral"Feishu"(BL 13) and "Xinshu"(BL 15). Rats of the Captopril group was treated by gavage of Captopril suspension (5 mg/mL, 25 mL/kg), and those of the M＋C group treated by the combined two methods. All the treatments were given once a day for 3 weeks. The general conditions and behaviors of rats were observed. The left ventricular mass index (LVMI) and right ventricular mass index (RVMI) were detected for assessing the cardiac performance. Morphological changes of myocardium were observed by HE staining. Enzyme linked immunosorbent assay (ELISA) was used to detect the concentrations of B-type natriuretic peptide (BNP) and precursor N-terminal pro-brain natriuretic peptide (NT-pro BNP) in the serum. The expression levels of Bcl-2, Bax, Fas and FasL of the left ventricle of heart were detected by Western blot. After modeling, the pathological changes of myocardium (as myocardial cell swelling with vacuoles, myocardial fibre breakage, etc.) were obvious, the LVMI, RVMI, serum BNP and NT-pro BNP concentrations, and myocardial Bax, Fas and FasL protein expression levels were significantly increased in the model group compared with the normal group (P<0.01), while the expression level of Bcl-2 was significantly down-regulated (P<0.01). Following the interventions, the myocardial injury was reduced, both LVMI and RVMI, serum BNP concentration and Bax, Fas and FasL expression levels in the three treatment groups, and serum NT-pro BNP concentration in the moxibustion and M＋C groups were significantly decreased (P<0.05, P<0.01), while the myocardial Bcl-2 protein levels in the three treatment groups were significantly increased relevant to the model group (P<0.01). Comparison among the three treatment groups showed that the effects of moxibustion ＋ Captopril were significantly superior to those of simple moxibustion and simple Captopril in suppressing CHF-induced increased expression of myocardial Bax, Fas and FasL, and in lessening CHF-induced decrease of Bcl-2 level (P<0.05, P<0.01). No significant differences were found among the three treatment groups in down-regulating LVMI and RVMI, and serum BNP content (P>0.05)．. Moxibustion can reduce myocardial injury and improve cardiac function in CHF rats, which may be related to its effects in down-regulating the expression of myocardial Bax, Fas and FasL proteins, and up-regulating the expression of Bcl-2 protein to inhibit cardiomyocyte apoptosis.

Prophylactic efficacy of patchoulene epoxide against ethanol-induced gastric ulcer in rats: Influence on oxidative stress, inflammation and apoptosis

Patchoulene epoxide (PAO), a tricyclic sesquiterpene isolated from the long-stored patchouli oil, has been demonstrated the anti-inflammatory activity in vivo based on our previous study. However, the gastric protective effect of PAO still remains unknown. Therefore, in the present study, ethanol-induced gastric ulcer model was carried out to evaluate the anti-ulcerogenic activity of PAO and to elucidate the potential mechanisms that involves. According to our results, macroscopic examination revealed that PAO could significantly reduce ethanol-induced gastric ulcer areas as compared with the vehicle group, which was also supported by the histological evaluation result. As for its potential mechanism, the anti-inflammatory activity of PAO contributed to gastric protection through reversing the imbalance between pro- and anti-inflammatory cytokines and modulating the expressions of NF-κB pathway-related proteins including p-IκBα, IκBα, p-p65 and p65. Besides, PAO was able to enhance the expressions of antioxidant enzymes including glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT), and down-regulate malonaldehyde (MDA), an indicator of lipid peroxidation. Furthermore, immunohistochemistry analysis exhibited potent anti-apoptosis effect of PAO, as evidence by down-regulating the protein expression of caspase-3, Fas and Fasl. Additionally, we also demonstrated that PAO could replenish PGE2 and NO mucosal defense. In conclusion, these findings suggested that PAO has gastric protective activity against ethanol and this might be related to its influence on inflammatory response, oxidative stress, apoptosis cascade and gastric mucosal defense.

Effects of harmaline on cell growth of human liver cancer through the p53/p21 and Fas/FasL signaling pathways.

The effects of harmaline on the viability and apoptosis of human liver carcinoma were investigated in vitro. HepG2 cells were treated with harmaline (0-10 µM), and the proliferation and apoptosis of HepG2 cells were investigated using an MTT assay and flow cytometry, respectively. The protein expression of cellular tumor antigen p53 (p53), cyclin-dependent kinase inhibitor 1 (p21), tumor necrosis factor receptor superfamily member 6 (Fas), Fas ligand (FasL) and caspase-8 was subsequently measured using western blotting. In addition, an ELISA was used to analyze caspase-8/3 activity. Harmaline significantly increased p53, p21, Fas and FasL protein expression in HepG2 cells. Additionally, treatment with harmaline significantly increased the expression of caspase-8 and caspase-8/3 activity. The results from the present study suggest that harmaline suppresses the viability, but induces the apoptosis, of human liver carcinoma cells through upregulation of the p53/p21 and Fas/FasL signaling pathways.

The effect of interleukin-17 on podocyte-associated factors and apoptosis and molecular mechanism

Objective To investigate the effect of interleukin(IL)-17 on podocyte-associated factors and apoptosis in podocytes and explore the molecular mechanism. Methods The podocytes were used as the object of study.The apoptosis of podocytes induced by IL-17 in a dose(1 nm/ml, 10 nm/ml, 50 nm/ml, 100 nm/ml) and time(12 h, 24 h, 48 h, 72 h) way and the protein expression of Fas and FasL of podocytes induced by IL-17 were detected by flow cytometrey.The podocytes were divided into the blank control group and 100 ng/ml IL-17 induction group.The expressions of mRNA of Nephrin, WT1, Synaptopodin, Podocylaxin, Fas and FasL were measured by Real-time RT PCR.The proteins of WT1, Caspases8, Caspases3 in podocyte were detected by immunocytochemistry method. Results IL-17 promoted the apopotosis of podocyte in a dose and time way(P 0.05). Conclusion IL-17 may cause renal injure by inducing the apoptosis of podocyte and decreasing the expression of Podocylaxin in podocyte. Key words: Interleukin-17; Podocyte; Apoptosis; Fas; Podocylaxin,

Effects of celecoxib on cell apoptosis and Fas, FasL and Bcl-2 expression in a BGC-823 human gastric cancer cell line.

Fas, which is an apoptotic-related protein, has an important role in cell apoptosis. Fas ligand (FasL) binds to Fas and activates apoptosis signal transduction. We previously demonstrated that the efficiency of celecoxib inhibited the proliferation and apoptosis of HT-29 colon cancer cell line. The BGC823 cell line was used as an experimental model to evaluate the potential role of celecoxib on gastric cancer cell apoptosis. Inhibitory effects of celecoxib on cell viability were determined by MTT assay. Cell apoptosis was evaluated by flow cytometric analysis and laser confocal microscopy. The results of the present study demonstrated that celecoxib inhibited the viability of BGC823 cells in a concentration- and time-dependent manner. Furthermore, the effect of BGC823 cells apoptosis was increased in a concentration-dependent manner. Western blotting was used to determine the protein expression levels of Fas, FasL, and B-cell lymphoma-2 (Bcl-2). During the celecoxib-induced apoptosis of BGC823 cells, celecoxib upregulated Fas expression and downregulated FasL and Bcl-2 expression in a concentration-dependent manner. These results suggest that celecoxib inhibited the growth and induced apoptosis of BGC823 gastric cancer cells by regulating the protein expression of Fas, FasL and Bcl-2.

Experimental study on the apoptosis of cervical cancer Hela cells induced by juglone through c-Jun N-terminal kinase/c-Jun pathway.

To study the regulatory effect and molecular mechanism of juglone on apoptosis of cervical cancer Hela cells. Cervical cancer Hela cells were cultured and treated with different dosages of juglone (10, 20, and 40μmol/L, respectively) and c-Jun N-terminal kinase (JNK) inhibitor SP600125 (10, 20, and 40μmol/L, respectively). Then cellular proliferative activity and the expression of JNK/c-Jun pathway molecule and apoptotic molecule in the cells were detected. After 6, 12, 18 and 24h of treatment, the value for proliferative activity of cells treated with juglone was significantly lower than that of control group (P<0.05), and the anti-proliferative effect was more significant as the treatment period and juglone dosage increased (P<0.05). The protein expressions of Bax, CytC, Fas, FasL, Caspase-3, p-JNK and p-c-Jun in cells treated with juglone were significantly higher than those of control group (P<0.05), and the protein expressions of Bax, CytC, Fas, FasL, Caspase-3, p-JNK and p-c-Jun increased more remarkably as the juglone dosage increased (P<0.05). In cells treated with 40μmol/L juglone and SP600125, the protein expressions of Bax, CytC, Fas, FasL and Caspase-3 were significantly lower than those of cells treated with 40μmol/L juglone (P<0.05), and the protein expressions of Bax, CytC, Fas, FasL and Caspase-3 reduced more remarkably as the SP600125 dosage increased (P<0.05). Juglone can increase the expression of apoptotic molecules in mitochondrial pathway and death receptor pathway by activating JNK/c-Jun pathway, thus inducing apoptosis of cervical cancer cells.

Athyrium multidentatum (Doll.) Ching extract induce apoptosis via mitochondrial dysfunction and oxidative stress in HepG2 cells

Athyrium multidentatum (Doll.) Ching (AMC), a unique and nutritious potherb widely distributed in china, has been extensively used in traditional Chinese medicine. Previous studies indicated that AMC extract exhibited antioxidant and antitumor properties. However, the chemical composition of AMC and molecular mechanism of AMC toxicity to HepG2 cells have not yet been elucidated. Hence, this study aimed to investigate the chemical compositions and the underlying mechanisms of the antiproliferative and apoptotic effects of AMC on HepG2. HPLC-MS analysis showed that AMC contain five compounds with chlorogenic acid accounting for 43 percent. Also, AMC strongly inhibited the cell growth and induced apoptosis and cell cycle arrest in HepG2 cells by significantly upregulating the protein expressions of Fas, Fas-L, Bax/Bcl-2, cyto-c, cleaved caspase-3, and PARP in a dose-dependent manner, which indicates AMC induces apoptosis in HepG2 cells through both intrinsic and extrinsic pathways. Moreover, AMC provoked the production of ROS, H2O2, and NO, modulating the PI3K/Akt, MAPK, NFκB and Nrf2 pathways and their downstream transcriptional cascades, ultimately evoked oxidative stress and apoptosis in HpeG2 cells. Further in vivo experiments demonstrated that AMC significantly suppressed the tumor growth, suggesting that AMC may be a novel promising agent for hepatocellular carcinoma treatment.

Dynamic changes of FasL protein expression in retinal tissue with aging in retinal degeneration mice

Background Retinal transplantation is a new approach to the treatment of retinal degeneration diseases, and how to avoid or reduce the immune rejection after transplantation is a problem.In experimental studies on retinal transplantation, C57BL/6 mice are often used as donors and rd mice serve as recipients.Studies showed that Fas ligand (FasL) protein induces the apoptosis of Fas+ inflammatory cells by FasL/Fas signal pathway, speculating that FasL protein is associated with immune rejection after transplantation. Objective The aim of this study was to investigate FasL protein expression change in retinas of C57BL/6 mice and rd mice with aging and reveal the immune characteristics of the mouse retinas. Methods The frozen sections of eyeball from C57BL/6 mice and rd mice at the age of postnatal (PN)-0 week, PN-1 week, PN-2 week, PN-3 week and PN-4 week were prepared.The expression of FasL protein in the mouse retinas was examined by immnofluorescense technique.Images were acquired by fluorescence microscope and analyzed semi-quantitively by software from laser scanning confocal microscope as the fluorescence intensity (FI). The results were compared among different strains of mice. Results The retina developed imperfectly in PN-1 week C57BL/6 mice and FasL protein was positively expressed in the whole retina.In PN-2, 3 and 4 week C57BL/6 mice, retinas finished the development with 10 layers, and retinal pigment epithelium (RPE), inner segment (IS), outer limiting membrane (OLM), outer plexiform layer (OPL), inner nuclear layer (INL), inner plexiform layer (IPL) and ganglion cell layer (GCL). Retinal structure and expression of FasL protein in the whole retina of rd mice in the PN-1 week were similar with C57BL/6 mice, however, ONL cells of rd mice were evidently decreased with aging.The RPE, OPL, INL, IPL and GCL expressed the FasL protein in PN-2, PN-3 and PN-4 week rd mice.The mean FI of FasL protein in the RPE layer was 184.199±16.747, 186.797±7.904 and 184.319±18.795 in rd mice of PN-2 week, PN-3 week and PN-4 week, which were significantly higher than 160.402±22.851, 160.995±22.799 and 105.787±17.676 in C57BL/6 mice (t=-3.360, P=0.002; t'=-4.277, P=0.000; t=-12.175, P=0.000). There were not significant differences in the mean FI of FasL protein in IPL between C57BL/6 mice and rd mice at ages of PN-2 week, PN-3 week and PN-4 week (all at P>0.05). Conclusions The cells of retinal ONL are gradually decreased with the development of rd mice, and FasL protein expression intensity in RPE is evidently enhanced in rd mice compared with C57BL/6 mice. Key words: Fas ligand protein; Retina/immunology; Pigment epithelium of eye/immunology; Apoptosis; Immune privilege; Retinitis pigmentosa; Mice, inbred, C57BL; rd mice

Molecular biological markers for proliferation, apoptosis, and angiogenesis in diffuse toxic goiter

to investigate clinical and laboratory parameters, as well as the expression of Ki-67, Bcl-2, Bax, Fas-L, CD34, VEGF, and FGF proteins in various postoperative outcomes of patients operated on for diffuse toxic goiter. The investigation enrolled 24 women who had undergone surgery using the technique described by E.S. Drachinskaya. Immunohistochemical tests were carried out according to the standard protocol. The expression of Ki-67, Bcl-2, Bax, Fas-L, CD 34, VEGF, angiopoietin, and FGF proteins was determined. The patients with postoperative thyrotoxicosis were ascertained to have a significantly greater expression of anti-apoptotic protein Bcl-2, proliferation marker Ki-67, vascular factors (FGF, VEGF), and CD 34. The relative expression area of the anti-apoptotic protein Bcl-2 of more than 2.19 or the proliferation protein Ki-67 of more than 1.059 was found to predict the development of postoperative thyrotoxicosis with an accuracy of higher than 85%.

临产前后胎盘上bcl-2、Fasl的表达与母血、羊水中ET-1、PGE&lt;sub&gt;2&lt;/sub&gt;浓度之间的关系

目的：探讨临产前后bcl-2、Fasl在胎盘滋养层细胞的蛋白表达变化及其与母血、羊水中内皮素-1 (Endothelin-1, ET-1)、前列腺素E2 (ProstaglandinE2, PGE2)的浓度之间的关系。方法：53例足月妊娠妇女分成两组，其中临产组28例，未临产组25例。用免疫组化(Immunohistochemical, IHC)法测定胎盘滋养层细胞bcl-2、Fasl的蛋白表达，用放射免疫法测定孕妇母血、羊水中ET-1、PGE2的浓度。结果：1) 临产组胎盘滋养层细胞bcl-2、Fasl的蛋白表达明显低于未临产组，两组平均灰度及阳性细胞率比较，差异有显著性(p 0.05)。结论：bcl-2、Fasl介导的胎盘滋养层细胞凋亡在启动分娩发动过程中具有重要作用；bcl-2的作用可能受ET-1、PGE2等细胞因子的调控。 Objective: To investigate the expressions of bcl-2 and Fasl proteins in placental trophoblastic cells on labor onset and their relationships with the concentrations of Endothelin-1 and Prostaglandin E2 in maternal blood and amniotic fluid. Methods: Fifty-three women on term pregnancy were divided into two groups. One group on the time of labor onset was twenty-eight women. Another group not on the time of labor onset was twenty-five women. Expressions of bcl-2 and Fasl proteins in placental trophoblastic cells were detected by immunohistochemical assay. The concentrations of ET-1 and PGE2 in maternal blood and amnionotic fluid were detected by radioimmune assay. Results: 1) Expressions of bcl-2 and Fasl proteins in the group of labor onset were significantly lower than that in the group of not labor onset. There was a statistically difference in the average gray scale and the ratio of positive cells between the two groups (p < 0.05). 2) The concentrations of ET-1 and PGE2 in maternal blood and amniotic fluid of labor onset group were higher than that of not labor onset group (p < 0.05). 3) There was a positive correlation between the concentrations of ET-1 and PGE2 in maternal blood and amniotic fluid, and the expression of bcl-2 proteins had a negative correlation with the concentrations of ET-1 and PGE2 respectively, but there was no correlation between the expression of Fasl proteins and the concentrations of ET-1 and PGE2. Conclusions: It was considered that the apoptosis of placental trophoblastic cells induced by bcl-2 and Fasl may play an important role in the initiation of term labor onset. ET-1 and PGE2 may control the expression of bcl-2.

Fluoride reduced the immune privileged function of mouse Sertoli cells via the regulation of Fas/FasL system

Previous investigations have demonstrated the adverse impacts of fluoride on Sertoli cells (SCs), such as oxidative stress and apoptosis. SCs are the crucial cellular components that can create the immune privileged environment in testis. However, the effect of fluoride on SCs immune privilege is unknown. In this study, mouse SCs were exposed to sodium fluoride with varying concentrations of 10−5, 10−4, and 10−3 mol/L to establish the model of fluoride-treated SCs (F-SCs) in vitro. After 48 h of incubation, F-SCs were transplanted underneath the kidney capsule of mice for 21 days, or cocultured with spleen lymphocytes for another 48 h. Immunohistochemical analysis of GATA4 in SCs grafts underneath kidney capsule presented less SCs distribution and obvious immune cell infiltration in F-SCs groups. In addition, the levels of FasL protein and mRNA in non-cocultured F-SCs decreased with the increase of fluoride concentration. When cocultured with F-SCs, lymphocytes presented significantly high cell viability and low apoptosis in F-SCs groups. Protein and mRNA expressions of FasL in cocultured F-SCs and Fas in lymphocytes were reduced, and the caspase 8 and caspase 3 mRNA levels were also decreased in fluoride groups in a dose-dependent manner. These findings indicated that fluoride influenced the testicular immune privilege through disturbing the Fas/FasL system.

The FAS-670 AA genotype is associated with high proviral load in peruvian HAM/TSP patients.

Human T-lymphotropic virus 1 (HTLV-1) is the etiologic agent of the HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). Apoptosis is a mechanism of defense elicited by many triggers, including cross-linking of the FAS receptor expressed in viruses-infected cells, and the ligand FASL presented by T-cytotoxic cells. As HAM/TSP has been associated with high levels of proviral load (PVL), we hypothesized that certain genotypes of single-nucleotide polymorphisms (SNPs) associated with a decreased protein expression of FAS and FASL could be risk factors for this disease. Three SNPs: FAS-670A/G (rs1800682), FAS-1377G/A (rs2234767), and FASL-844C/T (rs763110) were analyzed in 73 HAM/TSP patients and 143 HTLV-1 asymptomatic carriers. Ancestry informative markers were used to adjust for ethnicity through a principal component analysis. Gender, age, PVL, and the first three principal components were used as covariates. The FAS/FASL genotype distribution was not associated with HAM/TSP presence (P-> 0.05). The FAS-670 AA genotype was associated with high PVL in comparison to FAS-670 GG in HAM/TSP patients (P = 0.015), while in asymptomatic carriers low levels of PVL were observed (P > 0.05). Our findings suggest that rs1800682, rs2234767, and rs763110 genotypes are not associated with the presence of HAM/TSP, but that the FAS-670 AA genotype can promote higher PVL values in HAM/TSP patients. J. Med. Virol. 89:726-731, 2017. © 2016 Wiley Periodicals, Inc.

Fluid shear stress suppresses TNF-α-induced apoptosis in MC3T3-E1 cells: Involvement of ERK5-AKT-FoxO3a-Bim/FasL signaling pathways

TNF-α is known to induce osteoblasts apoptosis, whereas mechanical stimulation has been shown to enhance osteoblast survival. In the present study, we found that mechanical stimulation in the form of fluid shear stress (FSS) suppresses TNF-α induced apoptosis in MC3T3-E1 cells. Extracellular signal-regulated kinase 5 (ERK5) is a member of the mitogen-activated protein kinase (MAPK) family that has been implicated in cell survival. We also demonstrated that FSS imposed by flow chamber in vitro leads to a markedly activation of ERK5, which was shown to be protective against TNF-α-induced apoptosis, whereas the transfection of siRNA against ERK5 (ERK5-siRNA) reversed the FSS-medicated anti-apoptotic effects. An initial FSS-mediated activation of ERK5 that phosphorylates AKT to increase its activity, and a following forkhead box O 3a (FoxO3a) was phosphorylated by activated AKT. Phosphorylated FoxO3a is sequestered in the cytoplasm, and prevents it from translocating to nucleus where it can increase the expression of FasL and Bim. The inhibition of AKT-FoxO3a signalings by a PI3K (PI3-kinase)/AKT inhibitor (LY294002) or the transfection of ERK5-siRNA led to the nuclear translocation of non-phosphorylated FoxO3a, and increased the protein expression of FasL and Bim. In addition, the activation of caspase-3 by TNF-α was significantly inhibited by aforementioned FSS-medicated mechanisms. In brief, the activation of ERK5-AKT-FoxO3a signaling pathways by FSS resulted in a decreased expression of FasL and Bim and an inhibition of caspase-3 activation, which exerts a protective effect that prevents osteoblasts from apoptosis.

Effects of Shenqi Fuzheng injection on Fas/FasL protein expression levels in the cardiomyocytes of a mouse model of viral myocarditis.

The aim of the present study was to examine the effects of Shenqi Fuzheng injection (SFI) on Fas and FasL protein expression levels in the cardiomyocytes of mice with viral myocarditis (VMC) and to explore the underlying anti-apoptotic mechanisms. A total of 120 male BALB/c mice were randomly divided into five groups as follows: Blank control group, model group, ribavirin group, low-dose SFI group and high-dose SFI group. The VMC model was established by the injection of coxsackievirus group B type 3 and saline, ribavirin or SFI was administered 30 min later. Cardiac samples were harvested from mice in each group on days 3, 10 and 30. Apoptosis of cardiac cells was examined using terminal deoxynucleotidyl transferase dUTP nick-end labeling, and Fas and FasL protein expression levels were detected using immunohistochemistry. Myocardial apoptosis and Fas/FasL protein expression levels were significantly increased in the model group, as compared with the blank group (P<0.01), whereas the apoptotic index (AI) and Fas/FasL protein expression levels of cardiac cells in the high-dose SFI group were significantly decreased compared with those in the model group on day 10 (acute phase; P<0.01). The AI and Fas/FasL protein expression levels of cardiac cells in the low- and high-dose SFI groups were also significantly decreased on day 30 (chronic phase; P<0.01); however, no differences between the high- and low-dose groups were detected. In conclusion, SFI relieves VMC via the downregulation of Fas and FasL protein expression and the inhibition of cell apoptosis.

Cinnamic Acid (CINN) Induces Apoptosis and Proliferation in Human Nasopharyngeal Carcinoma Cells

Background/Aims: CINN is the main ingredient of the traditional Chinese medicine cinnamon. The purpose of the present study was to investigate the effects of CINN on the proliferation and apoptosis of NPC cells and to elucidate the underlying molecular mechanisms. Materials and Methods: CNE2 human NPC cells were treated with various CINN concentrations. The effects of CINN on the proliferation and apoptosis of CNE2 NPC cells were examined using the MTT assay and flow cytometric analysis. Additionally, western blotting was performed to analyze the expression of a number of cell cycle- and apoptosis-related proteins. Results: The proliferation of CNE2 cells was significantly inhibited after treatment with different CINN concentrations for various lengths of time. The inhibitory effect of CINN was concentration-and time-dependent. Flow cytometric analysis showed that 2 mmol/L CINN displayed a significant apoptosis-inducing effect. The western blot analysis results showed that KLF6, Fas-L, Bax, P53 and caspase-3 protein expression was drastically increased in the CNE2 cells after treatment with 2 mmol/L CINN, whereas Bcl-2 and cyclin D1 protein expression was markedly reduced. Conclusion: CINN inhibits the proliferation and induces the apoptosis of CNE2 cells. Therefore, CINN possesses a potential anti-tumor effect.

Fas-FasL expression and myocardial cell apoptosis in patients with viral myocarditis.

The aim of the current study was to investigate Fas and FasL expression and myocardial cell apoptosis in viral myocarditis patients. Human heart specimens were selected from patients who were autopsied between February 2012 and February 2015; of these, 25 patients were diagnosed with viral myocarditis. Another 15 cases with no diagnosis of myocarditis were selected for the control group. All tissue specimens were divided into two parts, one for reverse transcription-polymerase chain reaction analysis and the other for immunohistochemical and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) analyses. In situ detection of apoptosis was performed by the TUNEL method, which revealed that myocardial cells from the viral myocarditis group exhibited significant apoptosis, whereas no apoptotic cells were observed in the control group. The number of cells staining positive for Fas and FasL protein in the viral myocarditis group was significantly higher than that in the control group (P < 0.05). There was also a correlation between Fas and FasL protein expression levels and scores (r = 0.92, P < 0.05). The mRNA expression of Fas and FasL was significantly higher in the viral myocarditis group than in the control group (P < 0.05). In conclusion, the Fas-FasL system may be involved in the pathogenesis of viral myocarditis. Furthermore, cytotoxic T lymphocytes may mediate cardiac muscle cells apoptosis via Fas-FasL signaling, and thus participate in the pathogenesis of viral myocarditis.