TNF related apoptosis-inducing ligand and its receptors in ocular tumors

Most of the ocular tumors have poor prognosis, and they remain a difficult problem in the area of ophthalmology. With the rapid development of molecular biology and immunologic techniques and the deep research on ocular tumor related genes, it becomes possible to diagnose and treat malignant tumors from the molecular level. The tumor necrosis factor related apoptosis-inducing ligand (TRAIL), a member of the tumor necrosis factor (TNF) super family, is a promising candidate, either alone or in combination with established cancer therapies, since it can initiate apoptosis through the activation of their death receptors. The ability of TRAIL to selectively induce apoptosis of transformed, virus-infected or tumor cells but not normal cells promotes the development of TRAIL-based cancer therapy. Here, we will review TRAIL and its receptors' structure, function, mechanism of action and application in ocular tumors therapy.

Antitumor effect of tumor necrosis factor-related apoptosis inducing ligand combined with mevastatin on a human glioma cell line SWO-38

BACKGROUND： Previous studies have reported that statins are less toxic to the human body and have greater antitumor activity; however, few studies have addressed the antitumor effect of statins combined with tumor necrosis factor-related apoptosis inducing ligand （TRAIL）. OBJECTIVE： To explore the effect of TRAIL combined with mevastatin on the proliferation and apoptotic cell death of a human glioma cell line SWO-38, and to study its mechanism of action. DESIGN, TIME AND SETTING： An in vitro control experiment was performed at the Central Laboratory of the Third Hospital Affiliated to Sun Yat-sen University, between January and April 2009. MATERIALS： The human SWO-38 cell line was provided by Cell Research, Department of Animal Experimental Center of Sun Yat-sen University; human recombinant soluble TRAIL by R＆D, USA; and mevastatin by Sigma, USA. METHODS： SWO-38 cells were separately incubated in TRAIL （100, 200, 300, 400, and 500 tJg/L） and mevastatin （5, 10, 20, 30, and 40 pmol/L） for 72 hours. In addition, SWO-38 cells were incubated in TRAIL （300 μg/L）, mevastatin （30 μmol/L）, and a solution containing both TRAIL and mevastatin for 12, 24, 48 and 72 hours. MAIN OUTCOME MEASURES： Cell proliferation was detected using methyl thiazolyl tetrazolium assay; cell apoptosis was observed using Hoechst 33258 staining and fluorescence microscopy and was measured using Annexin V/propidium iodide flow cytometry; TRAIL R1/DR4 and TRAIL R2/DR5 protein expressions levels were measured using indirect immunofluorescence staining combined with flow cytometry in the recombinant soluble TRAIL （rsTRAIL, 300 tJg/L）, mevastatin （30 IJmol/L） and combination groups; TRAIL R1/DR4 and TRAIL R2/DR5 mRNA expression was detected using real-time polymerase chain reaction. RESULTS： rsTRAIL, mevastatin and their combination inhibited tumor proliferation in a time- and dose-dependent manner. The proliferation inhibitory rate and apoptosis rate of human SWO-38 cells in the combined group were significantly greater than the rsTRAIL or mevastatin alone group （P 〈 0.01）. TRAIL R1/DR4 and TRAIL R2/DR5 protein and mRNA expressions were increased in the combination group compared with mevastatin or rsTRAIL alone after 72 hours （P 〈 0.01）. CONCLUSION： Both rsTRAIL and mevastatin inhibit the proliferation and apoptosis of the human glioma cell line SWO-38, while their combination enhances the anti-tumor effect. The mechanism of action possibly correlates to the upregulation of TRAIL R1/DR4 and TRAIL R2/DR5 mRNA expression by mevastatin, thereby enhancing the cell sensitivity to rsTRAIL.

Expression of tumor necrosis factor related apoptosis inducing ligand receptor in glioblastoma

BACKGROUND： Receptors for tumor necrosis factor related apoptosis inducing ligand （TRAIL） include death receptor 4, death receptor 5, decoy receptor 1, and decoy receptor 2. Activation of death receptor 4 and 5 selectively kills tumor cells. OBJECTIVE： To detect TRAIL receptor expression in glioblastoma by immunohistochemistry and RT-PCR, and to compare this expression to that in normal brain tissue. DESIGN： Observational analysis. SETTING： Department of Pathology, the First Affiliated Hospital of Zhengzhou University; Henan Tumor Pathology Key Laboratory. PARTICIPANTS： Twenty-five patients （17 males and 8 females） who received glioblastoma resection were selected from the Fifth Affiliated Hospital of Zhengzhou University, between September 2003 to June 2004. All glioblastoma samples were diagnosed pathologically. Twenty patients （12 males and 8 females） with craniocerebral injury who received normal brain tissue resection were selected in the same time period. There were no significant differences in sex and age between glioblastoma patients or between craniocerebral injury patients （P 〉 0.05）. All patients and appropriate relatives provided informed consent, and this study was approved by the local research ethics committee. METHODS： Polyclonal antibody against TRAIL receptors and an immunohistochemical kit （batch number： 200502） were purchased from Boster Company, Wuhan. Immunohistochemistry： Expression of death receptor 4, death receptor 5, decoy receptor l, and decoy receptor 2 were observed in both glioblastoma and normal brain tissue. The experiment was performed according to the kit instructions, and positive staining was brown-yellow. Assessment： There were no positive signals （-）; weakly positive signals, positive cells 〈 25% （＋）; weakly positive signals, positive cells 25%-50% （＋＋）; strongly positive signals, positive cells 50%-75% （＋＋＋）; strongly positive signals, positive cells 〉 75% （＋＋＋＋）. Evaluation： Expression levels of TRAIL receptors were estimated in both normal brain tissue and glioblastoma. Expression of decoy receptor 1 and decoy receptor 2 mRNA in glioblastoma were detected by reverse transcription polymerase chain reaction, and expression of decoy receptor in glioblastoma was estimated. MAIN OUTCOME MEASURES： Comparison of death receptor and decoy receptor protein expression between glioblastoma and normal brain tissue; decoy receptor mRNA expression in glioblastoma. RESULTS： Death receptor protein expression was strongly positive （＋＋＋） in glioblastoma, while it was weakly positive （＋, ＋＋） in normal brain tissue. Therefore, expression rate of death receptor protein in the glioblastoma was significantly higher than that in the normal brain tissue （.~ 2 = 18.48, 23.03, P 〈 0.01）. Decoy receptor protein expression in the glioblastoma was significantly lower than that in the normal brain tissue （ x2 = 6.65, 18.76, P 〈 0.01）. The level of decoy receptor mRNA expression in glioblastoma was significantly higher than those of protein expression （ x 2 = 9.82, 10.09, P〈 0.01）. CONCLUSION： High expression of death receptor and low expression of decoy receptor are frequently observed in glioblastoma, suggesting that TRAIL receptor genes show an anti-tumor and expressive response during the initiation and development of the tumor. There are significant differences in decoy receptor expression between normal brain tissue and glioblastoma, suggesting that the restricted expression of decoy receptor in glioblastoma is regulated at the post-transcriptional level.

Tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis in glioma U87 cells

Studies have shown that tumor necrosis factor-related apoptosis-inducing ligand（TRAIL）exhibits strong induction of apoptosis in human glioma cells.It remains unclear whether the mitochondrion pathway,an important apoptosis signaling pathway,is involved in TRAIL-induced glioma cell apoptosis.In the present study,in vitro cultured human glioma U87 cells were treated with human recombinant soluble TRAIL.Apoptosis of glioma U87 cells,mitochondrial transmembrane potential（Δψm）,cytoplasmic cytochrome c concentration and changes in caspase-3,-8 and-9 activity following human recombinant soluble TRAIL treatment were investigated to determine the mechanism of glioma U87 cell apoptosis induced by TRAIL.Additionally,blocking caspase-8resulted in TRAIL-induced mitochondrion pathway activation,suggesting that TRAIL,through activating caspase-8,initiated a series of mitochondrial events and resulted in apoptosis of glioma U87 cells.

COMBINATION OF γ-INTERFERON WITH TRAIL AND CISPLATIN OR ETOPOSIDE INDUCES APOPTOSIS IN HUMAN NEUROBLASTOMA CELL LINE SH-SY5Y

Objective To study the effect of γ-interferon (IFNγ), tumor necrosis factor related apoptosis inducing ligand (TRAIL), and cisplatin or etoposide induced apoptosis in human neuroblastoma cell line SH-SY5Y and its possible molecular mechanisms.Methods The expressions of Caspase 8 mRNA and protein were detected with RT-PCR and Western blot analysis. The effects of IFNγ, TRAIL, IFNγ+TRAIL, IFNγ+Caspase 8 inhibitor+TRAIL, IFNγ+cisplatin+TRAIL, and IFNγ+etoposide+TRAIL on the growth and apoptosis of SH-SY5Y cells were detected with the methods of MTT and flow cytometry. The relative Caspase 8 activity was measured with colorimetric assay. Results Caspase 8 was undetectable in SH-SY5Y cells but an increased expression of Caspase 8 mRNA and protein was found after treatment with IFNγ. SH-SY5Y cells themselves were not sensitive to TRAIL, but those expressing Caspase 8 after treatment with IFNγ were. The killing effect of TRAIL on SH-SY5Y cells expressing Caspase 8 was depressed by Caspase 8 inhibitor. Cisplatin and etoposide could enhance the sensitivity of TRAIL on SH-SY5Y cells. The relative Caspase 8 activity of SH-SY5Y cells in IFNγ+TRAIL group was significantly higher than those of control group, IFNγ group, TRAIL group, and inhibitor group (P<0.01). There was no significant difference among IFNγ+TRAIL group, IFNγ+cisplatin+TRAIL group, and IFNγ+etoposide+TRAIL group.Conclusions IFNγ could sensitize SH-SY5Y cells to TRAIL-induced apoptosis and this may be realized by the upregulation of Caspase 8. Cisplatin and etoposide could enhance the killing effect of TRAIL on SH-SY5Y cells.

TRAIL-induced apoptosis of hepatocellular carcinoma cells is augmented by targeted therapies

AIM:To analyze the effect of chemotherapeutic drugs and specific kinase inhibitors,in combination with the death receptor ligand tumor necrosis factor-related apoptosis inducing ligand(TRAIL),on overcoming TRAIL resistance in hepatocellular carcinoma(HCC)and to study the efficacy of agonistic TRAIL antibodies,as well as the commitment of antiapoptotic BCL-2 proteins, in TRAIL-induced apoptosis. METHODS:Surface expression of TRAIL receptors (TRAIL-R1-4)and expression levels of the antiapoptotic BCL-2 proteins MCL-1 and BCL-xL were analyzed by flow cytometry and Western blotting,respectively. Knock-down of MCL-1 and BCL-xL was performed by transfecting specific small interfering RNAs.HCC cellswere treated with kinase inhibitors and chemotherapeutic drugs.Apoptosis induction and cell viability were analyzed via flow cytometry and 3-(4,5-Dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. RESULTS:TRAIL-R1 and-R2 were profoundly expressed on the HCC cell lines Huh7 and Hep-G2. However,treatment of Huh7 and Hep-G2 with TRAIL and agonistic antibodies only induced minor apoptosis rates.Apoptosis resistance towards TRAIL could be considerably reduced by adding the chemotherapeutic drugs 5-fluorouracil and doxorubicin as well as the kinase inhibitors LY294002[inhibition of phosphoinositol- 3-kinase(PI3K)],AG1478(epidermal growth factor receptor kinase),PD98059(MEK1),rapamycin(mam- malian target of rapamycin)and the multi-kinase inhibitor Sorafenib.Furthermore,the antiapoptotic BCL-2 proteins MCL-1 and BCL-xL play a major role in TRAIL resistance:knock-down by RNA interference increased TRAIL-induced apoptosis of HCC cells.Additionally, knock-down of MCL-1 and BCL-xL led to a significant sensitization of HCC cells towards inhibition of both c-Jun N-terminal kinase and PI3K.CONCLUSION:Our data identify the blockage of survival kinases,combination with chemotherapeutic drugs and targeting of antiapoptotic BCL-2 proteins as promising ways to overcome TRAIL resistance in HCC.

EGFR inhibitors sensitize non-small cell lung cancer cells to TRAIL-induced apoptosis

Apoptosis induced by tumor necrosis factor-related apoptosis-inducing ligand(TRAIL) can be regulated by the epidermal growth factor(EGF) signaling pathway.In this study,recombinant adenoviral vectors that encode TRAIL gene from the hTERT/RGD promoter(AdTRAIL) was combined with drugs including gefitinib,elotinib,and cetuximab that inhibit EGFR and the EGF signaling pathway in non-small cell lung cancer(NSCLC) cell lines to investigate their antitumor activity.In vitro,compared to single reagent,AdTRAIL combined with EGFR inhibitors reduced proliferation and enhanced apoptosis in H460,A549,and SW1573 cell lines.Western blot results suggested that these effects were relative to up-regulation of pro-apoptosis protein BAX and down-regulation of p-AKT.In vivo,AdTRAIL combined with cetuximab resulted in a significant growth reduction in H460 xenografts without damage to the main organs of nude mice.Histological examination and TUNEL analyses of xenografts showed that cetuximab enhanced cell apoptosis induced by AdTRAIL.These results indicate that EGFR inhibitors enhanced AdTRAIL anti-tumor activity in NSCLC cell lines and that inhibiting the AKT pathway played an important role in this enhancement.

Apoptosis and Expression of Protein TRAIL in Granulosa Cells of Rats with Polycystic Ovarian Syndrome

The relationship between apoptosis of granulosa cells and follicle development arrest in polycystic ovarian syndrome (PCOS) rats, and the contribution of tumor necrosis factor related apoptosis inducing ligand (TRAIL) in apoptosis of granulosa cells were explored. By using sodium prasterone sulfate rat PCOS model was induced. The apoptosis of granulosa cells in ovaries of rats was observed by TdT-mediated dUTP-biotin nick end-labeling (TUNEL), and the expression of TRAIL protein and mRNA in granulosa cells was detected by using immunhistochemical staining and reverse transcription polymerase chain reaction (RT-PCR) respectively. The apoptotic rate and the expression of protein TRAIL in granulosa cells were significantly higher in antral follicles from the PCOS rats than in those from the control rats (P<0.01, P<0.05). There was no significant difference in apoptotic rate and the expression of TRAIL protein in granulosa cells of preantral follicles between the PCOS rats and the control rats (P>0.05). No apoptosis and the expression of TRAIL protein in granulosa cells of primordial follicles were found in the two groups. The expression of TRAIL mRNA was significantly stronger in granulosa cells from the PCOS rats than in those from the con- trol rats (P<0.01). It was suggested that the apoptotic rate in granulosa cells was significantly higher in antral follicle from the PCOS rats than in those from the control rats. TRAIL played a role in regu- lating the apoptosis of granulosa cells in PCOS rats.

Induction of apoptosis in osteogenic sarcoma cells by combination of tumor necrosis factor-related apoptosis inducing ligand and chemotherapeutic agents

Background Osteosarcoma is one of the most common primary malignant tumors of bone with poor prognosis. TNF-related apoptosis inducing ligand （TRAIL） is a member of the tumor necrosis factor （TNF） cytokine family. TRAIL induces apoptosis in various tumor cell lines but is not found to be cytotoxic to many normal cell types in vitro. We investigated the cytotoxic activity of TRAIL and chemotherapeutic agents, including methotrexate （MTX）, doxorubicin （DOX） and cisplatin （CDDP）, on established osteosarcoma cell line-OS-732. Methods OS-732 cells were incubated with chemotherapeutic agents MTX,DOX and CDDP at various peak plasma concentrations（PPC）, 0.1PPC,1PPC and 10PPC, alone or with 100 ng/ml of TRAIL for 24 hours or 48 hours. MTT was used to evaluate the cytotoxic activity of different agents on OS-732. The apoptosis proportion was assayed by flow cytometry. Cellular morphologic changes were observed by phase contrast microscope, scan electron microscope, and transmission electron microscope. Results The inhibitory rate was （24.438±3.414）% with TRAIL of 100 ng/ml for 24 hours. The cells were responsive to DOX and CDDP with a dose-effect relationship （P〈0.05）. In OS-732 cells, DOX and CDDP cooperated synergistically with TRAIL when incubated the cells with them for 24 hours （the combined inhibitory rate is （58.360±2.146）% and （54.101±-2.721）%, respectively）. TRAIL alone or drugs alone induced the apoptosis rate was less than 25% （P〈0.05）. However, the combination of TRAIL and MTX did not present synergistic effects on OS-732 cells （P〉0.05, compared with TRAIL alone）. Conclusions Osteosarcoma OS-732 cells were not responsive to TRAIL-induced apoptosis. DOX and CDDP sensitize osteosarcoma OS-732 cells to TRAIL-induced apoptosis. The combination of TRAIL and MTX presented no synergistic effects on killing OS-732 cells.

Downregulation of cyclooxygenase-1 is involved in gastric mucosal apoptosis via death signaling in portal hypertensive rats

门高血压(PHT ) gastropathy 是肝肝硬化的经常的复杂并发症，带之一从肝硬化死亡引起。Apoptosis 广泛地被认为从坏死的房间死亡是房间死亡和一个不同实体的一个活跃精力依赖者模式。胃的 mucosal apoptosis 是否涉及 PHT gastropathy，是不清楚的。通过 cyclooxygenase (艇长) 生产的前列腺素(PG ) 被认为从损害和 apoptosis 在胃肠的 mucosa 的保护起一个关键作用。然而，在 PHT gastropathy 的艇长的角色仍然不清楚地被理解。这研究的目的是调查(1 ) 胃的 mucosal apoptosis 是否涉及 PHT gastropathy，(2 ) 艇长的 downregulation 贡献这 apoptosis。在这研究，当 mucosal 增长在 PHT 老鼠被禁止时，我们证明胃的 mucosal apoptosis 显著地被增加。胃的 mucosal COX-1 显著地在 mRNA 和蛋白质层次被压制，并且 PGE2 在 PHT 老鼠被减少。进一步， PGE2 处理在 PHT 老鼠压制了胃的 mucosal apoptosis。然而，胃的 mucosal COX-2 层次没在假冒操作老鼠和 PHT 老鼠之间不同。肿瘤坏死因素的胃的 mucosal 层次 --(TNF-) 并且船边交货 ligand，然而并非 TNF 相关的导致 apoptosis ligand，被增加，并且激活的 caspase-8 和 caspase-3 层次是在 PHT 老鼠的 upregulated。到 cytosol 的从线粒体的细胞色素 c 的版本没在 PHT 老鼠被观察。我们的数据显示 COX-1 的 downregulation 经由死亡涉及胃的 mucosal apoptosis 调停信号的类型 -- 我在 PHT 老鼠的房间死亡。

EGCG Enhances TRAIL-mediated Apoptosis in Human Melanoma A375 Cell Line

Tumor necrosis factor-related apoptosis-inducing ligand （TRAIL） is a promising anti-cancer agent. Epigallocatechin-3-gallate （EGCG） is a polyphenolic constituent of green tea. In this study, inhibitory effect of combined use of EGCG and TRAIL on human melanoma A375 cells was examined and the possible mechanism investigated. The cells were divided into 4 groups： control group, EGCG group （EGCG： 10, 20 μg/mL）, TRAIL group （TRAIL： 25 ng/mL） and EGCG＋TRAIL group （combined group）. The growth inhibition was measured in the A375 cells treated with different concentrations of TRAIL （（25, 50, 75, 100, 125, 150 ng/mL） by MTT assay. The apoptosis was assessed by flow cytometry. The expressions of DR4 and DR5 were detected by flow cytometry and western blotting. The activities of caspase-8 and caspase-3 were determined by colorimetric assay. The results showed that TRAIL could dose-dependently inhibit the growth of A375 cells and the IC50 of TRAIL was 150 ng/mL. The apoptosis rate was 11.8% in the TRAIL group, 5%–7% in the EGCG group and 48.9%–59.1% in the combined group. Significant difference was found in the apoptosis rate between the combined group and the EGCG or TRAIL group （P〈0.05 for each）. The expression of DR4 instead of DR5 was significantly increased in the EGCG group. The activity of caspase-3 rather than caspase-8 was substantially enhanced in the EGCG group. These results suggest that EGCG is useful for the TRAIL-based treatment for melanoma.

Antitumor and radiosensitization effect of 12C6+heavy-ion irradiation mediated by radiation-inducible gene therapy

Radio genetic therapy which combines gene therapy with radiotherapy has shown promising results in cancer treatment. In this study, an oncolytic adenovirusbased gene therapy system regulated by radiation was constructed to improve the cancer curative effect. This gene therapy system incorporated the radiation-inducible early growth response gene(Egr-1) promoter and the anticancer gene tumor necrosis factor-related apoptosis-inducing ligand(TRAIL). To confirm the antitumor effect of Ad-ET combined with^12C^(6+)tion irradiation, the survival and apoptosis fraction of tumor cells HT1080 and normal cells MRC-5 in combination treatment were detected by CCK-8 assay and FACS analysis. Then the expression levels of TRAIL gene and protein were tested by real-time PCR and western blotting. The results show that^12C^(6+)tion irradiation could induce cell growth inhibition and apoptosis by activating the TRAIL gene expression in tumor cells, while exhibiting no obvious toxicity to the normal lung cell line MRC-5. Theresults also demonstrate that use of an oncolytic adenovirusbased radiation-inducible gene therapy system together with^12C^(6+)tion irradiation could cause synergistic antitumor effect specifically in tumor cells but not in normal cells. The results indicate that the novel radio genetic therapy could potentiate radiation treatment by improving the safety and efficiency of monotherapy, and provide theoretical support for clinical application of combination treatment.

针刀调控线粒体途径软骨细胞凋亡防治大鼠膝骨关节炎

背景:针刀治疗膝骨关节炎的疗效确切,但其作用机制并不十分明确。目的:基于破骨细胞相关受体-肿瘤坏死因子相关的凋亡诱导配体-骨保护素(OSCAR-TRAIL-OPG)途径分析针刀对膝骨关节炎大鼠膝关节软骨细胞凋亡的影响。方法:采用随机数字表法将27只SD大鼠分为正常组(9只)、模型组(9只)、针刀组(9只),正常组大鼠常规饲养,不进行任何处理;模型组、针刀组采用膝关节内注射木瓜蛋白酶建立左后肢膝骨关节炎模型,造模成功后给予针刀组大鼠针刀干预,1次/周,共3次。干预结束后进行相关检测。结果与结论:①与正常组相比,模型组大鼠Lequesne MG行为学评分升高(P<0.01);与模型组相比,针刀组大鼠Lequesne MG行为学评分降低(P<0.01)。②苏木精-伊红染色显示与正常组相比,模型组大鼠膝关节软骨表面磨损且不平整,软骨细胞肿胀、破裂且数量减少,细胞排列杂乱;针刀组大鼠膝关节软骨表面较为平整,软骨细胞数量较多且排列较规整,结构基本清晰。③免疫组化染色显示与正常组相比,模型组大鼠膝关节软骨组织中OSCAR、TRAIL阳性表达增加(P<0.01),OPG阳性表达减少(P<0.01);与模型组相比,针刀组大鼠膝关节软骨组织中OSCAR、TRAIL阳性表达减少(P<0.01),OPG阳性表达增加(P<0.01)。④TUNEL染色显示与正常组相比,模型组软骨细胞凋亡数量增加(P<0.01);与模型组相比,针刀组软骨细胞凋亡数量减少(P<0.01)。⑤RT-qPCR与Western blot检测显示与正常组相比,模型组大鼠关节软骨组织中OSCAR、TRAIL、Bax表达升高(P<0.01),OPG、Bcl-2表达降低(P<0.01);与模型组相比,针刀组大鼠关节软骨组织中OSCAR、TRAIL、Bax表达降低(P<0.01),OPG、Bcl-2表达升高(P<0.01)。⑥针刀干预可减轻膝骨关节炎大鼠关节软骨组织损伤,该作用可能与OSCAR-TRAIL-OPG通路阻断线粒体途径凋亡信号释放有关。

T2DM患者血清sTWEAK、sCD40L水平与糖尿病微血管并发症的相关性分析

目的 探讨2型糖尿病(T2DM)患者血清可溶性肿瘤坏死因子(TNF)样凋亡弱诱导因子(sTWEAK)、可溶性CD40配体(sCD40L)水平与糖尿病微血管并发症(DMA)的相关性。方法 选取2021年3月-2023年3月航天中心医院收治的150例T2DM患者为研究对象,根据患者是否合并DMA分为DMA组(n=69)及T2DM组(n=81)。检测并比较两组患者血清sTWEAK、sCD40L水平及临床资料,采用Logistic回归分析影响T2DM并发微血管病变的危险因素,绘制受试者工作特征曲线(ROC)分析sTWEAK、sCD40L对T2DM并发微血管病变的预测价值。结果 DMA组患者血清sTWEAK、sCD40L水平均高于T2DM组(P<0.05);DMA组的病程、FPG及HbA1c、sTWEAK、sCD40L水平均高于T2DM,差异均有统计学意义(P<0.05);Logistic分析结果显示病程长、HbA1c高水平、sTWEAK高水平及sCD40L高水平均是导致T2DM并发微血管病变的独立危险因素(P<0.05)。ROC结果显示,血清sTWEAK、sCD40L单独及联合预测T2DM并发微血管病变的AUC(95%CI)分别为0.714(0.635~0.785)、0.744(0.666~0.812)、0.859(0.792~0.910),二者联合的预测效能优于单独检测(Z=2.626、2.395,P<0.05)。结论 血清sTWEAK、 sCD40L在DMA患者中均异常升高,与DMA关系密切,sTWEAK、sCD40L联合对于T2DM并发微血管病变的预测价值较高。

肝癌组织中TRAIL表达及其与癌细胞增殖、凋亡、耐药的相关性分析

目的:研究肿瘤坏死因子相关凋亡诱导配体(tumor-necrosis-factor-related apoptosis-inducing ligand, TRAIL蛋白在肝细胞癌(hepatocellular carcinoma, HCC)组织中的表达水平,分析其与临床病理特征及预后的相关性,并探讨TRAIL对HCC细胞凋亡、增殖及耐药的影响。方法 :利用免疫组织化学法检测186例HCC患者中HCC组织及癌旁肝组织TRAIL、Caspase-3、NF-κB、P-糖蛋白(P-glycoprotein, P-gP)及Ki-67的表达,分析TRAIL与临床病理特征及预后的关系,并探讨TRAIL与HCC生物学行为(增殖、凋亡、耐药)的相关性。结果:HCC组织中TRAIL表达明显低于癌旁肝组织(P<0.01)。HCC中Caspase-3、NF-κB、P-gP的阳性表达率分别为38.2%、84.4%、66.1%,Ki-67在HCC中的高表达率为75.8%,低表达率为24.2%。Spearman相关性分析显示TRAIL与Caspase-3的表达呈正相关(P<0.01),与NF-κB、P-gP、Ki-67的表达均呈负相关(均P<0.05)。Kaplan-Meier生存分析显示,TRAIL阳性表达患者的平均生存时间显著长于阴性表达者(P<0.01)。单因素和Cox模型多因素分析结果显示TRAIL低表达与术后复发是HCC预后的独立影响因素(P<0.05)。结论:TRAIL在HCC中低表达,并与脉管癌栓、肿瘤病理分级、术后复发有关,提示TRAIL低表达是HCC恶性进展中的关键性分子事件。检测TRAIL表达在评价HCC分级、浸润、复发及预后中有一定的价值。

全反式维A酸可促进肿瘤相关诱导配体对多种胰腺癌细胞的凋亡作用

目的:观察全反式维A酸(all-trans retinoic acid,ATRA)是否能促进肿瘤坏死因子相关凋亡诱导配体(tumor necrosis factor related apoptosis induced ligand,TRAIL)对多种胰腺癌细胞的凋亡作用。方法:将携带TRAIL基因的pCA13质粒分别转染SW1990、Patu8988和Bx-PC3等3种胰腺癌细胞,转染24 h后加入ATRA或等体积溶剂。四甲基偶氮唑盐(methyl thiazolyl tetrazolium,MTT)法测定细胞活性,流式细胞仪检测细胞的凋亡率和TRAIL受体R1、R2的表达,末端脱氧核苷酸转移酶(terminal deoxynucleotidyl transferase,TdT)介导的dUTP末端标记法(TdT mediated dUTP nick end labeling,TUNEL)和Hoechst双染色法、透射电镜观察细胞凋亡。结果:转染pCA13/TRAIL质粒后再加入ATRA,细胞生长活性与未加ATRA组相比抑制显著(P<0.05)。流式细胞仪检测发现加入ATRA较未加药组明显促进细胞凋亡(P<0.05)。TUNEL和Hoechst双染色法后透射电镜可观察到细胞凋亡表现。但检测TRAIL-R1、TRAIL-R2的表达发现加入ATRA与未加药组差异无统计学意义(P>0.05)。结论:ATRA可促进TRAIL对多种胰腺癌细胞的凋亡作用,其机制与TRAIL-R1、TRAIL-R2的上调无关。

血清可溶性肿瘤坏死因子相关凋亡诱导配体、半乳凝素3、摄食抑制因子-1与慢性阻塞性肺疾病合并抑郁的相关性分析

目的探讨血清中可溶性肿瘤坏死因子相关凋亡诱导配体(s TRAIL)、半乳凝素3、摄食抑制因子-1与慢性阻塞性肺疾病(COPD)合并抑郁的相关性。方法选取2019年7月至2021年5月河北北方学院附属第一医院收治的COPD急性加重期住院患者173例,其中单纯COPD组103例,COPD合并抑郁组70例,记录并比较两组临床资料及血清s TRAIL、半乳凝素3、摄食抑制因子-1水平。采用logistic回归模型分析COPD合并抑郁的影响因素,进一步绘制受试者操作特征(ROC)曲线评估预测效能。结果COPD合并抑郁组COPD评估测试评分、治疗依从性差占比、合并慢性病占比、独居占比及血清s TRAIL、半乳凝素3、摄食抑制因子-1水平均高于单纯COPD组,第1秒用力呼气量占用力肺活量百分率(FEV_(1)/FVC)、第1秒用力呼气容积占预计值百分率(FEV_(1)pred)、职工医疗保险占比均低于单纯COPD组,差异有统计学意义(P<0.05)。多因素分析结果显示,FEV_(1)/FVC、FEV_(1)pred、治疗依从性、医疗保险类型及血清s TRAIL、半乳凝素3、摄食抑制因子-1均为COPD合并抑郁的影响因素(P<0.05)。ROC曲线分析结果显示,血清s TRAIL、半乳凝素3、摄食抑制因子-1三者单独及联合均对COPD合并抑郁有一定的预测价值(P<0.05)。结论COPD患者发生抑郁与多种因素有关,血清s TRAIL、半乳凝素3、摄食抑制因子-1联合检测对COPD患者合并抑郁诊断有一定价值。

TRAIL联合顺铂诱导三阴乳腺癌MDA-MB-231细胞过度性自噬的分子机制

流行病学报道,乳腺癌成为目前女性发病率和死亡率最高的癌症[1]。而铂类药物也是临床上最常用的周期非特异性抗肿瘤药物,在临床治疗实体肿瘤中显示了良好的疗效[2]。其中顺铂作为一线临床的传统用药,其靶点选择范围窄、疗效差、易耐药等问题,尚未得到解决。肿瘤坏死因子相关凋亡诱导配体(tumor necrosis factor-related apoptosis-inducing ligand,TRAIL)属于肿瘤坏死因子(tumor necrosis factor,TNF)超家族。

芹菜素7-O-葡萄糖苷对抗TRAIL乳腺癌细胞凋亡的作用机制研究

[目的]探讨芹菜素7-O-葡萄糖苷(AGL)对抗肿瘤坏死因子相关凋亡诱导配体(TRAIL)乳腺癌细胞凋亡的影响。[方法]通过CCK-8法、蛋白免疫印迹、流式细胞法检测AGL、TRAIL、AGL+TRAIL对乳腺癌细胞活性、聚二磷酸腺苷核糖聚合酶-1(PARP-1)水平和细胞凋亡的影响,通过激光扫描共焦显微镜检测细胞ROS水平,采用ELISA法检测丙二醛(MDA)水平和超氧化物歧化酶(SOD)活性以及Western-blot检测PARP-1、p53、p53上调凋亡调制物(PUMA)水平。建立MCF7移植瘤模型,观察AGL+TRAIL处理下移植瘤生长和凋亡情况。[结果]TRAIL(50 ng/mL)+AGL(20、40μmol/L)处理时,MCF7细胞存活率降低(P<0.05),Cleaved PARP-1水平升高(P<0.05);TRAIL(50 ng/mL)+AGL(20μmol/L)处理时,MCF7细胞凋亡率、Cleaved-Cas-3、Cleaved-Cas-9水平均显著升高(P<0.05)。AGL(10、20μmol/L)处理时,MCF7细胞ROS(+)细胞数、MDA水平显著升高(P<0.05),SOD活性显著降低(P<0.05);TRAIL(50 ng/mL)+AGL(20μmol/L)MCF7细胞ROS(+)细胞数目、p53、PUMA、Cleaved PARP-1相对蛋白水平显著高于TRAIL(50 ng/mL)处理(P<0.05),TRAIL(50 ng/mL)+AGL(20μmol/L)+NAC(4 mmol/L)MCF7细胞ROS(+)细胞数、p53、PUMA、Cleaved PARP-1相对蛋白水平显著低于TRAIL(50 ng/mL)+AGL(20μmol/L)处理(P<0.05)。AGL+TRAIL组小鼠移植瘤质量、体积和肿瘤组织p53阳性率、PUMA3阳性率均显著低于Control组(P<0.05),细胞凋亡率显著高于Control组(P<0.05)。[结论]AGL联合TRAIL可促抗TRAIL MCF7细胞凋亡,可能与增强细胞氧化应激损伤和激活Caspase级联反应有关。

心肌梗死介入治疗后sTRAIL-R2表达与颈动脉斑块细胞凋亡及炎症反应的相关性

目的探究心肌梗死患者介入治疗后可溶性肿瘤坏死因子相关凋亡诱导配体受体2(sTRAIL-R2)表达与颈动脉斑块细胞凋亡及炎症反应的相关性。方法选择2021年1月至2022年5月该院收治的心肌梗死行介入治疗后患者102例作为研究对象,对其行颈动脉内膜切除术获取颈动脉斑块片段,根据sTRAIL-R2表达水平分为sTRAIL-R2高表达组和sTRAIL-R2低表达组。检测患者斑块组织Bax、半胱天冬氨酸蛋白酶(Caspase)-8、Caspase-3活性,CD45、CD68表达水平及白细胞介素(IL)-6、IL-10、C反应蛋白(CRP)、肿瘤坏死因子(TNF)-α和IL-1β水平,检测患者斑块组织细胞凋亡相关蛋白表达并分析sTRAIL-R2表达水平与患者斑块组织细胞凋亡、炎症反应的相关性。结果sTRAIL-R2高表达组患者斑块组织Caspase-8、Caspase-3活性,Bax、Caspase-3蛋白表达水平,CD45、CD68阳性细胞检出数,IL-6、IL-10、CRP、TNF-α、IL-1β水平均高于sTRAIL-R2低表达组,B淋巴细胞瘤-2基因(Bcl-2)蛋白表达水平低于sTRAIL-R2低表达组,差异均有统计学意义(P<0.05)。sTRAIL-R2表达水平与Caspase-8、Caspase-3活性,CD45、CD68、IL-6、IL-10、CRP、TNF-α、IL-1β水平与Bax、Caspase-3蛋白表达水平均呈正相关,与Bcl-2蛋白表达水平呈负相关(P<0.05)。结论sTRAIL-R2高表达可引起颈动脉粥样硬化斑块组织Caspase-8、Caspase-3活性升高,细胞凋亡相关蛋白表达水平上调,并引起斑块炎症反应加剧,可能导致易损斑块出现。