The effectiveness of serum S100B, TRAIL, and adropin levels in predicting clinical outcome, final infarct core, and stroke subtypes of acute ischemic stroke patients / Eficacia de los niveles séricos de S100B,TRAIL y adropina para predecir el resultado clínico, el núcleo del infarto final y los subtipos de ataque cerebrovascular de los pacientes con accidente cerebrovascular isquémico agudo

Introduction: More than half of all worldwide deaths and disabilities were caused by stroke. Large artery atherosclerosis is identified as a high etiological risk factor because it accounts for 20% of ischemic stroke. Objectives: To identify the significance of TRAIL and adropin release and the relative changes related to S100B levels, as well as the relationship between these biomarkers and the final infarct core, the clinical outcome, and the presence of large artery atherosclerosis in acute stroke patients. Materials and methods: Over a one-year period, demographic, clinical, and neuroimaging findings of 90 consecutive patients with acute ischemic stroke were evaluated. Results: The mean age of participants was 69.28 ± 10 and 39 patients were female. The increased level of S100B and the decreased levels of sTRAIL with adropin were significantly associated with moderate to severe neurologic presentation (p=0.0001, p=0.002, p=0.002, respectively). On the control CT, a large infarct core was significantly associated with decreased serum levels of sTRAIL and adropin (p=0.001 and p=0.000, respectively); however, the levels of S100B were not significantly associated with good ASPECTS score (p=0.684). Disability and an unfavorable outcome were significantly related to the decreased level of sTRAIL and adropin (p=0.001 and p=0.000 for THRIVE score>5, respectively). Decreased sTRAIL and adropin levels and an increased S100B level were correlated with the presence of large artery atherosclerotic etiologic factors (p=0.000, p=0.000, p=0.036, respectively). Conclusion: TRAIL and adropin serum levels were associated with poor clinical outcomes and greater infarcted area in acute ischemic stroke patients.

Introducción. Más de la mitad de todas las muertes y discapacidades en todo el mundo fueron causadas por accidentes cerebrovasculares. La aterosclerosis de las grandes arterias se identifica como un factor de alto riesgo etiológico debido a que representa el 20 % de los accidentes cerebrovasculares isquémicos. Objetivo. Determinar la importancia de la liberación de TRAIL y adropina y los cambios relativos relacionados con los niveles de S100B, así como la relación entre estos biomarcadores y el núcleo final del infarto, el resultado clínico y la presencia de aterosclerosis de arterias grandes en pacientes con accidente cerebrovascular agudo. Materiales y métodos. Durante un año, se evaluaron los hallazgos demográficos, clínicos y de neuroimágenes de 90 pacientes con accidente cerebrovascular isquémico agudo. Resultados. La edad media de los pacientes fue de 69,28 ± 10 y 39 eran mujeres. El aumento del nivel de S100B y la disminución de los niveles de sTRAIL y adropina se asociaron significativamente con una presentación neurológica moderada a grave en los pacientes (p=0,0001, p=0,002 y p=0,002, respectivamente). En la TC de control, un gran núcleo de infarto se asoció significativamente con una disminución del nivel sérico de sTRAIL y adropina (p=0,001 y p=0,000, respectivamente); sin embargo, los niveles de S100B no se asociaron significativamente con una buena puntuación en el ASPECT (p=0,684). La discapacidad y el resultado desfavorable se relacionaron significativamente con la disminución de los niveles de sTRAIL y adropina (p=0,001 y p=0,000 para una puntuación >5 en el THRIVE, respectivamente). La disminución de los niveles de sTRAIL y adropina y el aumento del nivel de S100B, se correlacionaron con la presencia de un factor etiológico aterosclerótico de arterias grandes entre la población de estudio (p=0,000, p=0,000 y p=0,036, respectivamente). Conclusiones. Los niveles séricos de TRAIL y adropina se asociaron con un resultado clínico deficiente y una mayor área infartada en pacientes con ataque cerebrovascular isquémico agudo.

The research of brain glioma therapy for nude mouse using transgenic human umbilical cord mesenchymal stem cells overexpressing TRAIL / 中国输血杂志

【Objective】 To explore the establishment methods of transgenic human umbilical cord mesenchymal stem cells (hUC-MSCs) overexpressing tumor necrosis factor(TNF)-related apoptosis-inducing ligand (TRAIL) based on the transposons, and attempt to apply it on the nude mice mode with glioma. 【Methods】 PiggyBac transposon system specially designed by us was used to prepare non-targeting and Her2-targeting hUC-MSCs that can stably express TRAIL through puromycin screening. The glioma cells expressing firefly luciferase (U87MG-LUC) were injected into the skull of the immunodeficient mice (BALB/c-nu/nu) with 1×106 cells per mouse. After 7 days of injection, the mice transplanted with U87MG were detected with a small animal living imager to determine the size and location of the tumors in skull. Then we injected the glioma-transplantation nude mouse with two kinds of transgenic hUC-MSCs expressing TRAIL (named as untarget-TRAIL and target-TRAIL, respectively), or the non-transgenic hUC-MSCs (all 1×106 cells per mouse) or PBS (named as WT-MSCs and PBS for negative control) respectively, and then monitored the changes of tumor signals by a small animal living imager every week for 3~4 weeks. 【Results】 After six passages to expand the cells, the both transgenic cell lines can stably express TRAIL gene. Their ratio of green fluorescent protein (GFP) positive cells can reach 93%-97%, and the positive ratio of their MSC-specific surface markers still maintained normal (CD34+, CD45+, and HLA-DR+ all <0.1%, CD90>99%, CD73>88%, and CD105 >60%). The median survival time (d) of U87MG-transplanted nude mice in the groups of untarget-TRAIL, target-TRAIL, WT-MSCs, and PBS was 41 vs 39 vs 24 vs 23(P<0.05). 【Conclusion】 The transgenic hUC-MSCs overexpressing TRAIL gene can significantly prolong the survival time of nude mice with brain glioma.

Influence of Tumor Necrosis Factor-related Apoptosis-inducing Ligand on Balance of Th17 and Treg Cells in Mice With Experimental Colitis / 胃肠病学

Background: Imbalance of Th17/Treg cells might play a key role in the initiation of Crohn's disease (CD). Moreover, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) pathway was suggested to be implicated in the pathogenesis of CD and other autoimmune diseases. Aims: To investigate the influence of TRAIL gene deletion on the balance of Th17 and Treg cells in mice with experimental colitis. Methods: TRAIL-/- C57BL/6 mice were obtained by CRISPR/Cas9 method. Then 20 TRAIL-/- mice and 20 wild-type (WT) mice were randomly divided into WT group, TRAIL-/- group, WT trinitrobenzenesulfonic acid (TNBS)-induced colitis group and TRAIL-/- TNBS-induced colitis group, with 10 mice in each group. The severity of colonic inflammation was assessed by disease activity index (DAI) and histology activity index (HAI). Peripheral blood and colonic tissue were collected to determine the expression levels of Th17 and Treg cell-associated transcription factors (RORγt, Foxp3) and cytokines [interleukin-17 (IL-17), IL-10] by real-time PCR, Western blotting and ELISA method, respectively. Results: Compared with untreated mice, mice with experimental colitis showed decreased body weight, shortened colon length as well as increased DAI and HAI (P<0.05), and the manifestations of colitis in TRAIL-/- mice were more serious than that in WT mice (P<0.05). In WT mice with experimental colitis, the expression levels of RORγt and IL-17 in peripheral blood and colonic tissue were significantly increased than those in untreated WT mice (P<0.05), while the expression levels of Foxp3 and IL-10 were significantly decreased (P<0.05). Furthermore, the expression levels of all four Th17 and Treg cell-associated molecules were higher in TRAIL-/- colitis mice than in WT colitis mice (P<0.05). In addition, the ratio of RORγt to Foxp3 were higher in TRAIL-/- colitis mice than in WT colitis mice (P<0.05). Conclusions: Deletion of TRAIL gene may aggravate the severity of colonic inflammation via up-regulating Th17/Treg ratio in mice with experimental colitis.

Effect of tumor necrosis factor-related apoptosis-inducing ligand gene knock-out on the gut microbiota in mice with dextran sodium sulphate induced colitis / 中华消化杂志

Objective@#To investigate the influence of tumor necrosis factor-related apoptosis-inducing ligant (TRAIL) deficiency on mice colitis and the gut microbiota composition by inclding the expermental colitis model in tumor necrosis factor-related apoptosis-inducing ligand gene knockout (TRAIL-/-) mice.@*Methods@#C57BL/6 TRAIL-/- mice and wild type (WT) mice were selected and assigned into TRAIL-/- control group (eight mice), TRAIL-/- colitis group (16 mice), WT control group (eight mice) and WT colitis group (16 mice). The mice of two colitis groups were oral administrated with 3.5% dextran sulphate sodium (DSS) in drinking water for seven consecutive days to induce experimental colitis model. The severity of colitis was evaluated by clinical appearance and histopathological examination. The colonic tissue samples of mice were collected and microbiota profile was analyzed by 16S rDNA sequencing method. USEARCH software and R language were used to analyze the difference of gut microbiota among TRAIL-/- control group, TRAIL-/- colitis group, WT control group and WT colitis group. T test and Mann-Whitney U test were used for statistical analysis.@*Results@#After modeling, the disease activity index (DAI) of WT colitis mice and TRAIL-/- colitis mice both gradually increased over time. Furthermore, compared with colitis mice, TRAIL-/- colitis mice developed body weight loss, diarrhea and hemafecia earlier. On the seventh day after modeling, the percentage of body weight loss of TRAIL-/- colitis mice and WT colitis mice was (28.98±2.84)% and (17.87±3.70)%, respectively; and the difference was statistically significant (t=9.53, P<0.01). The length of colon of TRAIL-/- colitis mice was shorter than that of WT colitis mice ((4.63±0.28) cm vs. (6.02±0.41) cm), and the difference was statistically significant (t=11.20, P<0.01). The DAI of TRAIL-/- colitis mice was higher than that of WT colitis mice (3.00±0.00 vs. 2.32±0.05), and the difference was statistically significant (t=54.40, P<0.01). The histological score of TRAIL-/- colitis mice was higher than that of WT colitis mice (6.19±0.25 vs. 3.87±0.22), and the difference was statistically significant (t=27.87, P<0.01). Under the microscope, colonic mucosal epithelial injury, crypt structure destruction and inflammatory cell infiltration were more obvious in TRAIL-/- colitis mice than in WT colitis mice. The alpha diversity of colonic flora was more significant in TRAIL-/- colitis group compared with that of WT colitis group. At the family level, the relative richness of Deferribacteraceae, Ruminococcaceae, Rikenellaceae, F16 and Paraprevotellaceae significantly increased in TRAIL-/- colitis group, but the relative richness of Enterococcaceae obviously reduced ((19.839±19.991)% vs. (7.224±11.241)%, (3.564±2.543)% vs.(2.861±3.821)%, (0.123±0.066)% vs. (0.068±0.049)%, (0.032±0.033)% vs. (0.006±0.011)%, (0.153±0.098)% vs. (0.062±0.054)% and (0.013±0.027)% vs. (0.054±0.121)%, respectively; U=51, 69, 53, 35, 49 and 69, respectively; P<0.01 and 0.05, respectively). In addition, at the genus level the relative richness of Oscillospira, Mucispirillum and Cytophaga in TRAIL-/- colitis group remarkably elevated, and the relative richness of Enterococcus significantly decreased ((2.363±2.147)% vs. (1.813±2.847)%, (19.839±19.991)% vs. (7.223±11.241)%, (0.104±0.153)% vs. (0.046±0.069)% and (0.076±0.049)% vs. (0.135±0.074)%, respectively; U=70, 51, 66 and 65, respectively; P <0.05 and 0.01, respectively).@*Conclusion@#TRAIL deficiency aggravate DSS-induced colitis, and increase the alpha diversity of colonic microbiota in colitis mice.

Ginsenoside Rg3 synergistically promotes apoptosis of lung cancer H358 cells with TRAILand its mechanism / 中国肿瘤生物治疗杂志

@# Objective: To investigate the effect of ginsenoside Rg3 combined with TRAIL on the apoptosis of lung cancer H358 cells and its possible mechanism. Methods: After the completion of cell culture, H538 cells were treated with TRAIL (0, 50, 100, 200 ng/ ml ) or Rg3 (0, 25, 50, 100 μmol/L) for 48 h, and the cells were grouped according to different treatments, namely control group, 50 μmol/LRg3 group, 100 ng/ml TRAILgroup and 50 μmol/LRg3+100 ng/ml TRAILgroup. The effects of Rg3 and/or TRAILon the proliferation of H358 cells were detected by MTT assay. The effects of Rg3 and/or TRAIL on the morphological changes of H358 cells were observed by DAPI staining. Theapoptosis of H358 cells in each group was detected by flow cytometry. The effects of Rg3 and/or TRAIL on the expressions of death receptor 5 (DR5) and caspase-8 in H358 cells were detected by WB. Results: Compared with the other groups, the proliferation of lung cancer H358 cells was significantly inhibited, while the apoptosis was significantly elevated in the 50 μmol/LRg3+100 ng/ml TRAILgroup (P<0.05).After color developing, cells in 50 μmol/LRg3+100 ng/ml TRAILgroup had nuclear shrinkage, chromatin condensation, increased fluorescence intensity, and late morphological changes such as saturation fragmentation. Compared with the other groups, the expression levels of DR5 and caspase-3 ,8 in the cells of 50 μmol/L Rg3+100 ng/ml TRAIL group were significantly increased (P<0.05). Conclusion: Ginsenoside Rg3 combined with TRAIL can synergistically inhibit the proliferation and induce apoptosis of lung cancer H358 cells. The mechanism may be related to the up-regulation of DR5 and caspase-8 by ginsenoside Rg3.

Effect of tumor necrosis factor-related apoptosis-inducing ligand gene knock-out on the gut microbiota in mice with dextran sodium sulphate induced colitis / 中华消化杂志

Objective To investigate the influence of tumor necrosis factor-related apoptosis-inducing ligant (TRAIL) deficiency on mice colitis and the gut microbiota composition by inclding the expermental colitis model in tumor necrosis factor-related apoptosis-inducing ligand gene knockout ( TRAIL-/-) mice. Methods C57BL/6 TRAIL-/-mice and wild type (WT) mice were selected and assigned into TRAIL-/-control group (eight mice), TRAIL-/-colitis group (16 mice), WT control group (eight mice) and WT colitis group (16 mice).The mice of two colitis groups were oral administrated with 3.5% dextran sulphate sodium (DSS) in drinking water for seven consecutive days to induce experimental colitis model .The severity of colitis was evaluated by clinical appearance and histopathological examination .The colonic tissue samples of mice were collected and microbiota profile was analyzed by 16S rDNA sequencing method.USEARCH software and R language were used to analyze the difference of gut microbiota among TRAIL-/-control group, TRAIL-/-colitis group, WT control group and WT colitis group .T test and Mann-Whitney U test were used for statistical analysis . Results After modeling, the disease activity index (DAI) of WT colitis mice and TRAIL-/-colitis mice both gradually increased over time .Furthermore, compared with colitis mice, TRAIL-/-colitis mice developed body weight loss, diarrhea and hemafecia earlier .On the seventh day after modeling , the percentage of body weight loss of TRAIL-/-colitis mice and WT colitis mice was (28.98 ±2.84)%and (17.87 ±3.70)%, respectively; and the difference was statistically significant (t＝9.53, P?0.01).The length of colon of TRAIL-/-colitis mice was shorter than that of WT colitis mice ((4.63 ±0.28) cm vs.(6.02 ±0.41) cm), and the difference was statistically significant (t＝11.20, P?0.01).The DAI of TRAIL-/-colitis mice was higher than that of WT colitis mice (3.00 ±0.00 vs.2.32 ±0.05), and the difference was statistically significant (t ＝54.40, P? 0.01).The histological score of TRAIL-/-colitis mice was higher than that of WT colitis mice (6.19 ±0.25 vs. 3.87 ±0.22), and the difference was statistically significant (t ＝27.87, P?0.01).Under the microscope, colonic mucosal epithelial injury , crypt structure destruction and inflammatory cell infiltration were more obvious in TRAIL-/-colitis mice than in WT colitis mice.The alpha diversity of colonic flora was more significant in TRAIL-/-colitis group compared with that of WT colitis group .At the family level, the relative richness of Deferribacteraceae, Ruminococcaceae, Rikenellaceae, F16 and Paraprevotellaceae significantly increased in TRAIL-/-colitis group, but the relative richness of Enterococcaceae obviously reduced ((19.839 ±19.991)%vs. (7.224 ±11.241)%, (3.564 ±2.543)% vs.(2.861 ±3.821)%, (0.123 ±0.066)% vs.(0.068 ± 0.049)%, (0.032 ±0.033)% vs.(0.006 ±0.011)%, (0.153 ±0.098)% vs.(0.062 ±0.054)% and (0.013 ±0.027)%vs.(0.054 ±0.121)%, respectively; U＝51, 69, 53, 35, 49 and 69, respectively; P? 0.01 and 0.05, respectively).In addition, at the genus level the relative richness of Oscillospira, Mucispirillum and Cytophaga in TRAIL-/-colitis group remarkably elevated , and the relative richness of Enterococcus significantly decreased ((2.363 ±2.147)% vs.(1.813 ±2.847)%, (19.839 ±19.991)% vs.(7.223 ± 11.241)%, (0.104 ±0.153)%vs.(0.046 ±0.069)% and (0.076 ±0.049)% vs.(0.135 ±0.074)%, respectively; U＝70, 51, 66 and 65, respectively; P ?0.05 and 0.01, respectively).Conclusion TRAIL deficiency aggravate DSS-induced colitis, and increase the alpha diversity of colonic microbiota in colitis mice .

Effects of TNF-related apoptosis-inducing ligand on cell apoptosis of pancreatic cancer / 中华胰腺病杂志

Objective To investigate the mechanism of TNF-related apoptosis-inducing ligand (TRAIL) promoting apoptosis of pancreatic cancer cells SW1990, Patu8988 and BxPC3. Methods Three kinds of pancreatic cancer cells SW1990, Patu8988 and BxPC3 were transfected with the pCA13 plasmid carrying TRAIL gene ( pCA13 TRAIL group) and the blank plasmid control ( pCA13 group) , respectively. The expression of TRAIL mRNA in transfected cells was detected by RT-PCR, and the expression of TRAIL protein was detected by Western blot. The apoptosis rate and expression of TRAIL receptor R1 and R2 were detected by flow cytometry. Apoptosis was detected by TUNEL and Hoechst double staining, and observed by electron microscopy. The expression of caspase-3 in transfected cells was detected by immunohistochemistry. Results SW1990, Patu8988 and BxPC3 cells can expresse TRAIL mRNA and protein within 24 h after transfection. The apoptotic rate at 24 h after transfection was (27. 30 ± 5. 14)％, (13. 52 ± 0. 95)％ and (31. 40 ± 8. 70)％,respectively, which was higher than that of pCA13 group [(10. 58 ± 1. 88)％,(8. 42 ± 0. 46)％ and (16.11 ±1.66)％], respectively. The expression rates of TRAIL-R1 were (61.37 ± 3.05)％,(42.10 ± 5. 11)％ and (36. 64 ± 4. 84)％, respectively, and the expression rates of TRAIL-R2 were (36. 20 ± 4. 83)％,(37. 26 ± 8. 46)％ and (24. 32 ± 3. 71)％, respectively,which were higher than those of pCA13 group except PATU8988 cells. Positivity rates of caspase-3 were ( 14. 64 ± 5. 35 )％, ( 9. 92 ± 5. 50 )％ and (16. 12 ± 6. 74)％, which were obviously higher than ( 3. 01 ± 1. 50 )％, ( 1. 75 ± 0. 50 )％ and ( 3. 79 ± 1. 58)％ in pCA13 group,and the differences were statistically significant(P<0. 05). Conclusions TRAIL could up-regulate the expression of TRAIL R1 and R2 in multiple pancreatic cancer cell lines in vitro, and thus promote cell apoptosis.

Involvement of Up-regulation of Death Receptors and Bim in Hispolon-mediated TNF-related Apoptosis-inducing Ligand Sensitization in Human Renal Carcinoma

BACKGROUND: Hispolon has been shown to possess antitumor effects in various cancer cells. However, the underlying mechanisms are not fully understood. In this study, we evaluated the sensitizing effect of hispolon on TNF-related apoptosis-inducing ligand (TRAIL)-mediated apoptosis in human renal carcinoma cells. METHODS: Apoptosis was analyzed by using cell-based cytometer. The mRNA levels were assessed by reverse transcription-PCR. Bax activation was determined by oligomerization and fluorescence-activated cell sorting with Bax-NT monoclonal antibody. The protein expression was measured by Western blotting. RESULTS: Hispolon induced up-regulation of Bim and death receptors expression at the post-translational level. CONCLUSIONS: Hispolon enhanced TRAIL-mediated apoptosis in renal carcinoma cells, but not in normal cells.

Therapeutic Effects of Synthetic Antimicrobial Peptides, TRAIL and NRP1 Blocking Peptides in Psoriatic Keratinocytes / 전남의대학술지

Psoriasis is a chronic, recurrent, heterogeneous, cutaneous inflammatory skin disease for which there is no cure. It affects approximately 7.5 million people in the United States. Currently, several biologic agents that target different molecules implicated in the pathogenic processes of psoriasis are being assessed in diverse clinical studies. However, relapse usually occurs within weeks or months, meaning there is currently no cure for psoriasis. Therefore, recent studies have discovered diverse new potential treatments for psoriasis: inhibitors of bacteria such as Staphylococcus aureus, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and neuropilin 1 (NRP1). A promising approach that has recently been described involves modifying antimicrobial peptides to develop new cutaneous anti-bacterial agents that target inflammatory skin disease induced by Staphylococcus. Increased expression of TRAIL and its death receptors DR4 and DR5 has been implicated in the pathogenesis of plaque psoriasis. In addition, TRAIL has the ability to inhibit angiogenesis by inducing endothelial cell death and by negative regulation of VEGF-induced angiogenesis via caspase-8-mediated enzymatic and non-enzymatic functions. Since NRP1 regulates angiogenesis induced by multiple signals, including VEGF, ECM and semaphorins, and also initiates proliferation of keratinocytes through NF-κB signaling pathway in involved psoriatic skin, targeting NRP1 pathways may offer numerous windows for intervention in psoriasis. In this review, we will focus on the current knowledge about the emerging role of synthetic antimicrobial peptides, TRAIL and NRP1 blocking peptides in the pathogenesis and treatment of psoriasis.

The progress of TWEAK and its receptor Fn14 signal pathway in lung diseases / 中国医师杂志

Tumor necrosis factor-like attenuated apoptosis inducer (TWEAK) is a new member of the tumor necrosis factor super family.TWEAK regulates cellular proliferation,differentiation,migration,apoptosis and inflammation by binding its receptor,fibroblast growth factor-inducible 14 (Fn14),through the interaction between cells or in a paracrine fashion.The role of TWEAK / Fn14 signaling pathway in the development of lung diseases such as lung injury,asthma and lung cancer has drawn increasing attention.

Imperatorin enhances anti-tumor effect of TRAIL on breast cancer by upregulating expression of DR5 on cell surface / 中国病理生理杂志

AIM:To investigate the synergistic effect of imperatorin on enhancing the anti -tumor effect of TNF-related apoptosis-inducing ligand(TRAIL)on breast cancer and the mechanisms.METHODS:T-47D and MCF-7 breast cancer cells were divided into control group ,imperatorin group,TRAIL group,imperatorin+TRAIL group and imperatorin+TRAIL+death receptor 5(DR5)siRNA group.The viability of T-47D and MCF-7 cells was measured by MTT assay. The apoptosis and mitochondrial membrane potential in T-47D cells were analyzed by flow cytometry.Western blot and flow cytometry analysis were performed to evaluate the expression of DR 5 on T-47D cell surface and the activation of caspase-8 and caspase-3.RESULTS:Imperatorin significantly enhanced the inhibition of cell viability induced by TRAIL of T -47D and MCF-7 cells,and significantly increased the apoptosis of T-47D cells induced by TRAIL.Imperatorin treatment ob-viously induced upregulation of DR5 expression and production of reactive oxygen species in the T-47D cells.In addition,imperatorin enhanced the TRAIL-induced damage of mitochondrial membrane potential and activation of caspase -8 and caspase-3.CONCLUSION:Imperatorin enhances the anti-tumor effect of TRAIL on breast cancer via upregulating the ex-pression of DR5.

Targeting androgen receptor and trail: a novel treatment paradigm for breast cancer / 中国药理学与毒理学杂志

OBJECTIVE TNF- related apoptosis- inducing ligand(TRAIL)is a promising cancer therapeutic agent due to its minimal toxicity to normal tissues and remarkable apoptotic activity in tumors. However, most breast cancer cells are resistant to TRAIL- induced apoptosis. Our objectives are to investigate the underlying molecular mechanisms and to develop strategies to overcome such resistance. METHODS To identify modulators of TRAIL-induced apoptosis, we carried out a genome wide siRNA screen. To validate the screening result, we either silenced or overexpressed the identified genes in various breast cancer cells and changes in growth and TRAIL-induced cell apoptosis were determined in vitro and in an orthotopic xenograft mouse model. Finally, we investigated whether small molecules targeting the identified genes improve the effectiveness of TRAIL-therapy. RESULTS We unexpectedly identified androgen receptor (AR) to be responsible for TRAIL resistance. While AR is classically viewed as the key factor in prostate cancer progression, we found that AR expression levels were markedly elevated in human invasive breast cancer specimens including triple- negative breast cancers (TNBC) that are highly aggressive with poor prognosis. Importantly, breast cancer cell lines express different levels of AR that correlated with their TRAIL resistance. AR overexpression in MDA- MB- 231 and MDA- MB- 436 cells suppressed the TRAIL sensitivity whereas knockdown of AR rendered MCF-7 and MDA-MB-453 cells sensitive to TRAIL-induced apoptosis. AR overexpression also induced TRAIL resistance in breast tumors in vivo. Further, we observed an upregulation of the TRAIL receptor, death receptor 5 (DR5) in breast cancer cells, following the removal or inhibition of AR by its antagonists Casodex and MDV3100. Treatment with AR antagonists also enhanced TRAIL- induced breast cancer cell apoptosis. CONCLUSION AR signaling suppresses TRAIL-induced breast cancer cell apoptosis, in part, by suppressing DR5 expression, and a combination of AR antagonists together with TRAIL may be a novel and effective therapy for TNBC.

Effects of Combination of SAHA and TRAIL on Proliferation of Triple-negative Breast Cancer Cell MDA-MB-231 / 中国医科大学学报

Objective To investigate the effects of combined treatment of suberoylanilide hydroxamic acid(SAHA)and TNF?related apoptosis inducing ligand(TRAIL)on proliferation and morphology change of triple?negative breast cancer cell MDA?MB?231. Methods The effects of combination treatment of SAHA and TRAIL on proliferation and morphology change of MDA?MB?231 cells were monitored by RTCA. Morphology changes of MDA?MB?231 cells by different treatment factors were observed through time?lapse live cell imaging acquisition. Results Real?time cell proliferation assays showed that a synergistic effect were found when MDA?MB?231 cells were treated with combination of SAHA and TRAIL , and reached the best effect with 5μmol/L SAHA and 50 ng/mL TRAIL. The results of time?lapse live cell imaging acquisition showed that the growth inhibition of MDA?MB?231 cells with combined treatment of SAHA and TRAIL were more obvious than that with treatment of SAHA or TRAIL alone. Conclusion The combined treatment of SAHA and TRAIL induces a synergistic effect on growth inhibition in triple?negative breast cancer cell line MDA?MB?231.

Effects of combination with SAHA and TRAIL treatment on cell growth of er positive breast cancer cell line MCF-7 / 中国药学杂志

OBJECTIVE: To study the effects of suberoylanilide hydroxamic acid (SAHA) and TRAIL treatment on cell proliferation and apoptosis for ER positive breast cancer cell line MCF-7. METHODS: Human breast cell lines (MCF-7) were evaluated for the expressions of cell viability, cell apoptosis and cell cycle by muse cell analyzer. The mRNA levels of related apoptotic factors in MCF-7 cells were detected by real time PCR and solid phase apoptosis antibody microarray. RESULTS: After the combination with SAHA and TRAIL, the ability of cell proliferation and cell viability were depressed, and the cell apoptosis was induced. The cell cycle assay showed that the MCF-7 cells were arrested in G0/G1 phase with SAHA and TRAIL treatment. CONCLUSION: The combinatorial treatment of SAHA and TRAIL has a significantly inhibitory effect on cell growths of ER positive breast cancer MCF-7 cell.

Changes of sTWEAK and IL-6 levels in patients with acute coronary syndrome and its clinical signifi-cance / 心血管康复医学杂志

Objective:To explore diagnostic value of soluble tumor necrosis factor - like weak inducer of apoptosis (sTWEAK) and interleukin (IL) -6 concentrations in patients with acute coronary syndrome (ACS) .Methods:A total of 170 ACS patients hospitalized in our hospital from Jan 1st ,2014 to Jun 31st ,2014 were selected as ACS group ,mean‐while ,80 inpatients with stable angina pectoris (SAP) confirmed by coronary angiography (CAG) or coronary CT were se‐lected as SAP group ,and another 80 patients excluded for coronary heart disease (CHD) by CAG were regarded as normal control group .ACS group was further divided into ST elevation myocardial infarction (STEMI) group (n=45) ,non -STEMI (NSTEMI) group (n=52) and unstable angina pectoris (UAP) group (n=73) .Plasma sTWEAK and serum IL‐6 concentrations were compared among all groups .Results:Compared with normal control group and SAP group ,there were significant rise in concentrations of plasma sTWEAK [ (120.32 ± 10.15) ng/L vs .(123.86 ± 15.23) ng/L vs .(140.05 ± 17.51) ng/L] and serum IL‐6 [ (110.34 ± 26.01) pg/ml vs .(112.38 ± 25.74) pg/ml vs .(245.23 ± 68.58) pg/ml] (P0.05. There were no significant difference in plasma sTWEAK and serum IL‐6 concentrations among UAP group ,NSTEMI group and STEMI group , P>0.05 all .Conclusion:Plasma sTWEAK and serum IL‐6 concentrations significantly rise in ACS patients ,which possesses certain diagnostic value for ACS .

Effect of TRAIL on Expression of Multidrug Resistance Gene GST-π in Drug-resistant Human Gastric Cancer Cell Line SGC7901/VCR / 胃肠病学

Background:Multidrug resistance of tumor cells is one of the important factors that cause failure of chemotherapy in advanced gastric cancer. Tumor necrosis factor-related apoptosis-inducing ligand(TRAIL)may enhance the killing effect of chemotherapeutics on tumor cells,and reverse drug-resistant cell lines to sensitive cell lines,but its mechanism is not yet clear. Aims:To study the effect of TRAIL on expression of multidrug resistance gene glutathione S-transferase-π(GST-π)in drug-resistant human gastric cancer cell line SGC-7901 / VCR and the potential mechanism of TRAIL in reversing multidrug resistance of gastric cancer cells. Methods:SGC-7901 / VCR cells were treated with TRAIL in different doses (50,100,200 and 400 μg/ L)for 48 hours. After treatment,expression of GST-π mRNA in SGC-7901 / VCR cells and concentration of GST-π in culture supernatant were detected by RT-PCR and ELISA,respectively. Results:TRAIL could inhibit mRNA expression and protein secretion of GST-π in SGC-7901 / VCR cells in a dose-dependent manner within a certain range(≤200 μg/ L). The relative expression levels of GST-π mRNA in 50,100,200 and 400 μg/ L TRAIL groups were 0. 89 ± 0. 04,0. 77 ± 0. 08,0. 65 ± 0. 06 and 0. 61 ± 0. 03,respectively,and the concentrations of GST-π in culture supernatant in these groups were(57. 56 ± 1. 19)ng/ mL,(56. 30 ± 0. 80)ng/ mL,(31. 41 ± 1. 65)ng/ mL and (30. 80 ± 1. 34)ng/ mL,respectively,all were significantly lower than those in control group[1. 01 ± 0. 13 and(58. 62 ± 1. 38)ng/ mL,P all < 0. 05]. Conclusions:TRAIL may play a potential role in reversing multidrug resistance of gastric cancer cells through down-regulating GST-π expression.

Expressions and clinical significances of TRAIL and its receptors in non-small cell lung cancer / 国际肿瘤学杂志

Objective To investigation the expressions of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and its receptors in non-small cell lung cancer (NSCLC) and their clinical significances.Methods The serum expression levels of TRAIL in 79 cases of NSCLC and 80 cases of normal subjects were detected by enzyme-linked immunosorbent assay (ELISA).The expressions of TRAIL-R2 and TRAIL-R4 in 42 cases of NSCLC and matched normal tissues were detected by immunohistochemistry.The relationships among TRAIL,TRAIL-R2,TRAIL-R4 and clinicopathologic features of NSCLC were analyzed.Results The expression of TRAIL in NSCLC patients was lower than that in normal human [(994.3 ±293.0)ng/ml vs.(1 141.7 ±266.1)ng/ml,t =3.29,P =0.00].The expression of TRAIL was closely correlated with clinical stage (F =2.28,P =0.00) and differentiated degree (t =5.76,P =0.00).The positive expression rate of TRAIL-R2 in NSCLC was 73.8％ (31/42),significantly lower than that in the normal tissue 100.0％ (42/42) (x2 =3.88,P =0.05).The expression of TRAIL-R2 was closely correlated with clinical stage (x2 =27.89,P=0.00) and differentiated degree (x:=9.50,P =0.00).The positive expression rate of TRAIL-R4 in NSCLC was 81.0％ (34/42),significantly higher than that in the normal tissue 50.0％ (21/42) (x2 =7.34,P =0.01).The expression of TRAIL-R4 was also closely correlated with clinical stage (x2 =17.82,P =0.00) and differentiated degree (x2 =4.47,P =0.03).There was a negative correlation between the expression of TRAIL-R2 and TRAIL-R4 in NSCLC (r =-0.67,P=0.01).Conclusion The decrease of TRAIL and TRAIL-R2 and increase of TRAIL-R4 expression may promote the occurrence and development of NSCLC,and they may provide targets for clinical treatment of NSCLC.

Isoproterenol Enhances Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand-Induced Apoptosis in Human Embryonic Kidney Cells through Death Receptor 5 up-Regulation

BACKGROUND AND OBJECTIVES: Chronic impairment of beta-adrenergic receptor signaling increases cardiac apoptosis, hypertrophy and fibrosis. The aim of this study was to investigate whether isoproterenol (ISO), an agonist of the adrenergic receptor, can enhance tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis in human embryonic kidney (HEK) 293 cells. MATERIALS AND METHODS: HEK 293 cells were treated with ISO and/or TRAIL for 24 hours. Cell viability was evaluated by microscopy and an established viability assay, and apoptotic cell death was analyzed by staining with fluorescein isothiocynate-annexin-V/propidium iodide (PI) and caspase activation. To confirm the mechanism of cell death induced by co-treatment with ISO and TRAIL, expression of TRAIL receptor 2 (death receptor 5, DR5) was evaluated by immunoblotting. RESULTS: Although ISO or TRAIL treatment decreased HEK 293 cell viability by 13% and 17%, respectively, co-treatment with ISO and TRAIL resulted in a markedly higher death rate of 35% after 24 hours. Increases were evident in early apoptotic cells (i.e., annexin-V positive/PI negative; 19.4%), late apoptotic cells (i.e., annexin-V positive/PI positive; 6.3%) and dead cells (i.e., annexin-V negative/PI positive; 1.1%) when cells were co-treated with ISO and TRAIL, compared to cells treated with either ISO or TRAIL. In addition, marked increases of cleaved cas-3, cleaved poly (adenosine diphosphate-ribose) polymerase and DR5 were observed in HEK 293 cells co-treated with ISO and TRAIL. CONCLUSION: Treatments combining ISO with TRAIL may be responsible for death of HEK 293 cells through DR5 up-regulation. Activation of adrenergic receptors is responsible for the synergistic cell death observed with TRAIL.

The expression of tumor necrosis factor -related apoptosis -inducing ligand (TRAIL)protein and it′s receptor 3,4 in colorectal cancer tissue and its significance / 中国基层医药

Objective To investigate the potential roles of tumor necrosis factor -related apoptosis -indu-cing ligand(TRAIL)signaling system in the pathogenesis of colorectal cancer.Methods Immunohistochemistry tech-niques were used,the TRAIL and its receptor(TRAILR3,TRAILR4)protein were analyzed in both 41 colorectal cancer samples(colorectal cancer group)and normal samples beside cancer tissue(normal group).Results The lev-els of TRAIL and TRAILR3 protein expression in the colorectal cancer group were significantly lower than those in the normal group[(0.237 ±0.036)vs.(0.289 ±0.069);(0.226 ±0.052)vs.(0.281 ±0.068),t =4.125,4.025,all P 0.05).The expression levels of them in the colorectal cancer group with poorl differentiation were notably lower than those with high -moderate differentiation[(0.205 ±0.021 )vs (0.245 ± 0.034);(0.185 ±0.032)vs (0.236 ±0.051),t =4.025,2.664,P 0.05].Moreover,the positive rate and expression levels of TRAILR4 protein was non -statistic significant difference between the two groups[93% vs 98%;(0.196 ±0.085)vs (0.219 ±0.061),χ2 =1.051,t =1.353,all P >0.05],and it was also non -significantly correlated with cancer cell differentiation and lymph nodes metastasis[(0.176 ±0.052)vs (0.201 ±0.091);(0.194 ±0.054)vs (0.197 ±0.100),t =0.667, 0.448,all P >0.05].Conclusion The levels of TRAIL and TRAILR3 expression are attenuated at colorectal cancer tissue.The expression of them are correlated with cancer cell differentiation grade.These findings indicate that TRAIL system may be associated with the malignant phenotype in colorectal cancer.

Inducing effect of recombinant mutant human TRAIL on apoptosis of lung cancer cells and its molecular mechanism / 肿瘤

Objective: To observe the apoptosis-inducing effect of recombinant mutant human tumor necrosis factor-related apoptosis-inducing ligand (rmhTRAIL): on human lung cancer cell line A549 and to identify the different protein expression profiles related to apoptosis. Methods: After the A549 cells were treated with rmhTRAIL, the proliferation viability was assessed by MTT assay, and the cell cycle and apoptosis were analyzed by flow cytometry. Then the liquid chromatograph-tandem mass spectrometer (LC-MS/MS): was used to detect the different expression profiles related to apoptosis. Finally, real-time fluorescence quantitative PCR (RFQ-PCR): and Western blotting were performed to identify the expression levels of apoptosis-related genes. Results: After A549 cells were treated with different concentrations of rmhTRAIL for 24 h, the inhibitory rates of proliferation were significantly increased in a concentration-dependent manner (all P < 0.05). The half maximal inhibitory concentration was (9.5±3.9): ×105 ng/mL. The early apoptosis rate was significantly increased after 5×105 ng/mL rmhTRAIL treatment for 24 h (P < 0.01). However, rmhTRAIL had no effect on cell cycle. After treatment with rmhTRAIL, the expressions of tumor necrosis factor receptor superfamily (TNFRSF): members 1 OB and 1 OD were down-regulated by 21.3% and 31.8%, respectively; and the expression of cellular FLICE-inhibitory protein (c-FLIP): was up-regulated by 2.867 times. Both RFQ-PCR and Western blotting analysis indicated that the expression of death receptor 5 (DR5), one of TNFRSF members, was down-regulated in both mRNA and protein levels (both P < 0.05). Relatively, the expression of c-FLIP was up-regulated in both mRNA and protein levels (both P < 0.05). Conclusion: rmhTRAIL can induce apoptosis of human lung cancer A549 cells, and this mechanism is associated with the down-regulation of DR5 expression and up-regulation of c-FLIP expression.