TRAIL受体DR5、DcR1在子宫内膜癌中的表达及临床意义

目的：探讨肿瘤坏死因子相关凋亡诱导配体（TRAIL）受体DR5、DcR1在子宫内膜癌中的表达及临床意义。方法：采用免疫组化Elivision^TM plus法检测DR5、DcR1在10例正常子宫内膜、12例增生性子宫内膜及42例子宫内膜癌中的表达。结果：子宫内膜癌中DR5的阳性表达率为36／42（85．71％）,明显高于正常子宫内膜3／10（30．00％）,（P〈0．01）。正常子宫内膜中DcR1的阳性表达率为100．00％,明显高于子宫内膜癌19／42（45．24％）,（P〈0.01）。在子宫内膜癌中,DR5的阳性表达与病理分级及肌层浸润呈反比。结论：子宫内膜癌中DR5的高表达使TRAIL用于子宫内膜癌的治疗在理论上具有可行性;子宫内膜癌中DcR1的表达可能导致TRAIL诱导的凋亡耐受。     

TRAIL及其受体DR5、DcR1在宫颈癌中的表达

目的:探讨TRAIL及其受体DR5、DcR1在宫颈癌中的表达及其临床意义.方法: 应用免疫组化SP法对10例正常宫颈组织、18例宫颈上皮内瘤变(CIN)组织和40例宫颈癌组织中TRAIL、DR5及DcR1的表达情况进行检测,并结合患者年龄、肿瘤分化程度、组织类型、有无淋巴结转移等临床病理因素进行分析.结果: TRAIL及DcR1在正常宫颈组织中的表达高于宫颈癌组织;宫颈癌组织中,TRAIL阳性表达随组织学分级降低而降低;DR5在正常宫颈、CIN和宫颈癌组织中的表达呈递增趋势.结论: TRAIL、DR5及DcR1的异常表达在宫颈癌变过程中起一定的作用.     

TRAIL及其受体DR4、DcR1在大肠癌及癌旁组织中的表达

 目的研究人肿瘤坏死因子相关凋亡诱导配体(TNF-related apoptosis inducing ligand,TRAIL)及其受体DR4、DcR1在大肠癌和癌旁组织中的表达及意义。方法采用免疫组化SP法检测42例大肠癌及其癌旁5cm组织、25例正常大肠粘膜组织中TRAIL及其受体DR4、DcR1表达水平。结果TRAIL及DR4在大肠癌、癌旁组织及正常大肠粘膜组织中的表达呈递增趋势,而DcR1的表达则与之相反(P<0.05);TRAIL和DR4在中、低分化癌中的表达明显低于高分化癌中的表达,而DcR1的表达则与之相反(P<0.01);TRAIL、DR4、DcR1的表达与肿瘤的病理类型、Duke′s分期及淋巴结转移与否等因素无关(P>0.05)。结论TRAIL、DR4、DcR1可能与大肠癌的发生、发展密切相关;DR4可能在TRAIL诱导的凋亡通路中发挥一定作用,而DcR1的表达与否一定程度上决定了TRAIL能否发挥其生物效应。     

TRAIL及其受体在乳腺癌组织中的表达及意义

目的探讨肿瘤坏死因子相关凋亡诱导配体（TRAIL）及其受体（DR4、DR5、DcR1、DcR2）在乳腺癌组织的表达及意义。方法用反转录聚合酶链反应（RT-PCR）法检测60例乳腺癌及对应的正常乳腺组织中TRAIL及其受体的表达。结果乳腺癌组织中TRAIL、DcR1、DcR2表达均低于正常乳腺组织(P＜0.05),且TRAIL及其受体的表达与乳腺癌的分期、分级等无关(P＞0.05)。结论TRAIL及其受体在乳腺癌凋亡过程中起着重要的作用,TRAIL及其诱骗受体低表达同乳腺癌的发生关系密切。     

TRAIL受体在人肝细胞癌中的表达及TRAIL诱导的肝癌细胞凋亡

目的 研究了RAIL受体在肝癌组织及肝癌细胞中的表达,探讨肿瘤坏死因子相关凋亡诱导配体(TRAIL)及其受体在肝癌细胞凋亡诱导中的作用。 方法 应用RT-PCR法和免疫组化方法检测肝癌组织及细胞系SK-Hepl中TRAIL受体的表达,并以TRAIL蛋白和γ-干扰素作用于SK-Hepl细胞,观察其对细胞的凋亡诱导作用。 结果 在肝癌组织及肝癌SK-Hepl细胞系中均可检测到TRAIL受体,其中,死亡受体DR4、DR5在肝癌、癌旁组织以及SK-Hepl肝癌细胞系中均呈高表达,而诱骗受体DcR1及DcR2呈低表达,明显低于正常组织。TRAIL对肝癌细胞有凋亡诱导作用,与干扰素的协同时,其作用明显增强。 结论 肝癌组织中存在着TRAIL受体的表达,TRAIL可诱导肝癌SK-Hepl细胞系凋亡,其作用与DR4、DR5的高表达有关。     

TRAIL受体DR5 、DcR1在子宫内膜癌中的表达及临床意义

目的:探讨肿瘤坏死因子相关凋亡诱导配体(TRAIL)受体DR5 、DcR1在子宫内膜癌中的表达及临床意义.方法:采用免疫组化ElivisionTM plus法检测DR5 、DcR1在10例正常子宫内膜、12例增生性子宫内膜及42例子宫内膜癌中的表达.结果:子宫内膜癌中DR5的阳性表达率为36/42(85.71%),明显高于正常子宫内膜3/10 (30.00%),(P<0.01).正常子宫内膜中DcR1的阳性表达率为100.00%,明显高于子宫内膜癌19/42(45.24%),(P<0.01).在子宫内膜癌中,DR5的阳性表达与病理分级及肌层浸润呈反比.结论:子宫内膜癌中DR5的高表达使TRAIL用于子宫内膜癌的治疗在理论上具有可行性;子宫内膜癌中DcR1的表达可能导致TRAIL诱导的凋亡耐受.     

Survivin、TRAIL在血管瘤中的表达

目的　探讨存活素 (Survivin)、肿瘤坏死因子相关凋亡诱导配体 (TRAIL)在血管瘤组织中的表达。方法　应用免疫组化的方法检测血管瘤组织、血管畸形中Survivin和TRAIL的表达。结果　Survivin血管瘤增生期、退化期和血管畸形中阳性表达率分别为76.9% ( 2 0 /2 6)、3 5 .7% ( 5 /14 )和 2 5 % ( 4 /16) (P <0 .0 5 )。卡方检验 :χ2 =12 .62 ,三者之间差异存在显著性意义 (P <0 .0 1) ,两两之间比较 ,增生期与消退期之间有差异 (P <0 .0 5 ) ,消退期与血管畸形之间无差异 (P >0 .0 5 ) ,增生期与血管畸形之间有差异 (P <0 .0 5 ) ,血管瘤与血管畸形之间有差异 (P <0 .0 5 )。TRAIL在增生期和消退期血管瘤组织中阳性表达率分别为 2 3 .1% ( 6/2 6)、78.6% ( 11/14 ) ,血管畸形中全部为阴性 ,卡方检验 :χ2 =9.3 1,三者之间差异存在显著性意义 (P <0 .0 1) ,两两之间比较 ,增生期与消退期之间有差异 (P <0 .0 5 ) ,消退期与血管畸形之间有差异 (P <0 .0 5 ) ,增生期与血管畸形之间无差异 (P >0 .0 5 ) ,血管瘤与血管畸形之间有差异。结论　Survivin和TRAIL的表达水平与血管瘤的增生、消退有关 ,Survivin可抑制内皮细胞的凋亡 ,促进血管瘤组织增生 ;TRAIL可通过诱导内皮细胞凋亡发挥作用 ,促进血管瘤消退     

肿瘤细胞对TRAIL敏感性与其表面DR5表达水平的相关性研究

目的　探讨肿瘤细胞表面DR5表达水平与其对肿瘤坏死因子相关的凋亡诱导配体(TRAIL)敏感性之间的关系。方法　利用抗DR5特异性单克隆抗体 ,采用流式细胞仪技术直接检测不同肿瘤细胞系表面DR5的表达水平 ,并采用TRAIL凋亡检测试剂盒检测肿瘤细胞对TRAIL诱导凋亡的敏感性 ,研究两者之间的关系。结果　不同肿瘤细胞表面DR5的表达水平分别为 :U937细胞97.9%、Jurkat细胞 95 .1%、SW4 80细胞 93.8%、HCT116细胞 86 .2 %、HL 6 0细胞 6 4 .2 %、HeLa细胞4 6 .6 %、K5 6 2细胞 13.1% ;TRAIL诱导的细胞凋亡率分别为 :U937细胞 72 .6 %、Jurkat细胞 85 .2 %、SW4 80细胞 78.6 %、HCT116细胞 70 .2 %、HL 6 0细胞 6 0 .1%、HeLa细胞 4 5 .4 %、K5 6 2细胞 12 .3%。经统计学分析 ,两者之间呈现非常明显的正相关 (r=0 .997,P <0 .0 0 1)。结论　肿瘤细胞对TRAIL的敏感性与其表面DR5表达水平有关 ,表明DR5的表达水平在TRAIL诱导细胞凋亡方面起着十分重要的作用     

Embelin 通过上调死亡受体 DR4 / DR5 mRNA 表达增加 HL - 60 细胞对 TRAIL的敏感性

目的：探讨Embelin增加HL-60细胞对肿瘤坏死因子相关凋亡诱导配体（TRAIL）的敏感性及可能的作用机制.方法：TRAIL 1、5、10、50及100 ng/ml分别处理HL-60细胞6、12、24及48h,MTT法绘制细胞生长曲线;TRAIL 10 ng/ml±亚细胞毒浓度的Embelin处理HL-60细胞6、12、24及48h,Annexin V/PI复染流式细胞术检测细胞凋亡,Western blot检测24h时Caspase-3、8及9的表达;亚细胞毒浓度的Embelin处理HL-60细胞6、12、24及48h,实时荧光定量PCR检测DR4及DR5 mRNA 的表达.结果：TRAIL对HL-60 细胞具有增殖抑制作用.亚细胞毒浓度的Embelin联合10 ng/ml TRAIL作用于HL-60细胞时细胞凋亡率较单独应用TRAIL时增加,相应的Caspase-3、8及9的表达也随之增加.结论：亚细胞毒浓度的Embelin可以增加HL-60细胞对TRAIL的敏感性,其机制可能与上调DR4及DR5 mRNA表达并进而启动凋亡途径有关.     

TRAIL及其受体在直肠癌中的表达及意义

目的：探讨人肿瘤坏死因子相关凋亡诱导配体（TNF-related apoptosis inducing ligand,TRAIL）及其受体DR4、DR5、DcR1、DcR2在直肠癌及癌旁组织中的表达及意义.方法：采用免疫组化EliVision法检测37例直肠癌及其癌旁组织和10例正常直肠粘膜组织中TRAIL及其受体DR4、DR5、DcR1、DcR2表达水平.结果：TRAIL在正常直肠粘膜组织、癌旁及直肠癌组织中的表达呈递减趋势,而DR4、DR5的表达则与之相反（P＜0.05）;TRAIL在中、低分化癌中的表达与高分化癌中的表达无差异,而DR4、DR5、DcR1、DcR2在中、低分化癌中的表达高于高分化癌中的表达（P＜0.05）;TRAIL及其受体的表达与肿瘤的病理类型、患者的年龄、性别等因素无明显相关性（P＜0.05）.结论：TRAIL在直肠癌中表达较正常直肠粘膜组织减弱,DcR1、DcR2在直肠癌中的表达高于DR4、DR5,提示TRAIL及其受体可能与直肠癌的发生、发展密切相关,为临床应用提供了一定的理论依据.     

Enhancement of TRAIL/Apo2L-mediated apoptosis by adriamycin through inducing DR4 and DR5 in renal cell carcinoma cells

Renal cell carcinoma (RCC) is one of the most drug-resistant malignancies in humans. We show that adriamycin (ADR) and TNF-related apoptosis-inducing ligand (TRAIL)/Apo2L have a synergistic cytotoxic effect against RCC cells. This synergistic cytotoxicity was obtained in ACHN, A704, Caki-1 and Caki-2 human RCC cell lines and freshly derived RCC cells from 6 patients. This synergistic effect, however, was not achieved in 5 samples of freshly isolated normal kidney cells. We further explored the mechanisms underlying this synergistic effect and found that the synergistic cytotoxicity of TRAIL/Apo2L and ADR was realized by inducing apoptosis. Sequential treatment with ADR followed by TRAIL/Apo2L induced significantly more cytotoxicity than the reverse treatment. ADR increased the expression of DR4 and DR5 in RCC cells, but not in the normal kidney cells. Furthermore, the synergistic cytotoxicity was significantly inhibited by DR4:Fc and DR5:Fc fusion proteins, which inhibit TRAIL/Apo2L-mediated apoptosis. In addition, caspase activity assays and treatment of caspase inhibitors demonstrated that the combination treatment with ADR and TRAIL/Apo2L activated caspase cascade, including caspase-9, -8, -6 and -3, which were the downstream molecules of death receptors. These findings indicate that ADR sensitizes RCC cells to TRAIL/Apo2L-mediated apoptosis through induction of DR4 and DR5, suggesting that the combination therapy of TRAIL/Apo2L and ADR might be effective for RCC therapy.     

Reactivation of death receptor 4 (DR4) expression sensitizes medulloblastoma cell lines to TRAIL

Object Apoptosis, a key cellular response to therapeutic agents is often inactivated in tumor cells. In this study, we evaluated the expression of the tumor necrosis family of death receptors, DR4 and DR5, in medulloblastoma tumor samples and cell lines to determine if epigenetic modulation of gene expression could sensitize tumor cell lines to TRAIL-mediated apoptosis. Methods Human medulloblastoma samples and cell lines were analyzed for DR4 and DR5 expression by quantitative PCR and immunofluorescence assays. Cell lines with downregulated expression of one or both genes were treated with the histone deacetylase inhibitor, MS-275, and the expression of DR4 and DR5 measured by quantitative PCR, Western blotting, flow cytometry and chromatin immunoprecipitation assays. Induction of apoptosis in the presence of MS-275 was evaluated by TUNEL assay and its ability to augment TRAIL-mediated cytotoxicity was determined by MTT assays, Western blotting and flow cytometry. Results Compared to normal cerebellum, DR4, but not DR5 expression was consistently downregulated in medulloblastoma tumor samples and in Daoy and D283 cell lines. Interestingly, MS-275 decreased cell growth and induced apoptosis in Daoy and D283 cells. In Daoy cells, this coincided with increased histone H3 and H4 acetylation at the DR4 promoter and enhanced DR4 gene and protein expression as well as elevated Caspase-8 activity. The involvement of DR4 in the cellular response to MS-275 was further confirmed by the observation that knockdown of DR4 and FADD abrogated apoptosis. Further, addition of TRAIL to MS-275 treated cells resulted in an enhancement of apoptosis, suggesting that the upregulated death receptors were functional. Conclusion Our study provides an understanding of the role of DR4 in apoptosis of medulloblastoma cell lines and suggests a potential contribution of aberrant histone deacetylation to the resistance of medulloblastoma cells to therapeutic death. This work was supported in part by Award P50CA127001 from the National Cancer Institutes and Grants from the National Brain Tumor Foundation and the American Cancer Society-IRG to VG.     

Oligomycin A enhances apoptotic effect of TRAIL through CHOP‐mediated death receptor 5 expression

Development of resistance to TNF‐related apoptosis‐inducing ligand (TRAIL) in tumor cells is one of the important problems in cancer treatment. Despite the previous report demonstrating that oligomycin suppressed TNF‐induced apoptosis, in our screening of small molecules enhancing cancer cell death to TRAIL, oligomycin A (OMA) was found to enhance TRAIL‐induced apoptosis in HeLa cells. CCAAT/enhancer‐binding protein homologous protein (CHOP) was found to directly bind to death receptor 5 (DR5) promoter through endoplasmic reticulum stress (ER‐stress) signaling and sensitize the cells to TRAIL. Among ER‐stress associated proteins, OMA triggered the inositol‐requiring enzyme 1 (IRE1) signaling pathway, leading to X‐binding protein 1 (XBP1) splicing, CHOP expression and DR5 upregulation. In contrast, small‐interfering RNA (siRNA) of CHOP reduced the number of apoptotic cells in response to the co‐treatment of TRAIL and OMA. Collectively, our data suggest that OMA enhances apoptotic death of cervical cancer cells to TRAIL through upregulation of CHOP‐mediated DR5 expression following ER‐stress. © 2011 Wiley Periodicals, Inc.     

TRAIL及其受体与卵巢恶性肿瘤的研究进展

肿瘤坏死因子相关的凋亡诱导配体(TNF-related apoptosis-inducing ligand,TRAIL)是最近发现的TNF家族的新成员,TRAIL有两类受体,一类是死亡受体,诱导肿瘤细胞或转化细胞凋亡,另一类是"诱骗"受体,保护正常细胞免遭TRAIL的诱导凋亡作用。卵巢癌患者经几个疗程化疗后,常对化疗药产生多药耐药现象,从而使化疗失败,最终导致肿瘤复发和转移。TRAIL和化疗药联合应用能逆转卵巢癌细胞的多药耐药性(multidrug resistance)。本文就TRAIL、TRAIL受体(TRAILR)表达及TRAIL的凋亡途径等在卵巢恶性肿瘤治疗中的应用做一全面综述。     

Induction of proapoptotic antibodies to triple-negative breast cancer by vaccination with TRAIL death receptor DR5 DNA

TNF-related apoptosis-inducing ligand receptor 2 [TRAIL-R2 or death receptor 5 (DR5)] is expressed at elevated levels in a broad range of solid tumors to mediate apoptotic signals from TRAIL or agonist antibodies. We tested the hypothesis that DR5 DNA vaccination will induce proapoptotic antibody to trigger apoptosis of tumor cells. BALB/c mice were electrovaccinated with DNA-encoding wild-type human DR5 (phDR5) or its derivatives. Resulting immune serum or purified immune IgG induced apoptosis in triple-negative breast cancer (TNBC) cells, which were also TRAIL sensitive. The proapoptotic activity of immune serum at dilutions of 0.5-2% was comparable to that of 1-2 μg/ml of TRAIL. Apoptotic activity of immune serum was enhanced by antibody crosslinking. Apoptotic cell death induced by anti-DR5 antibody was shown by the cleavage of PARP and caspase-3. In contrast, immune serum had no effect on the proliferation of activated human T cells, which expressed low levels of DR5. In vivo, hDR5 reactive immune serum prevented growth of SUM159 TNBC cells in severe combined immune-deficient mice. DR5-specific IFN-γ-secreting T cells were also induced by DNA vaccination. Furthermore, the feasibility to overcome immune tolerance to self DR5 was shown by the induction of mouse DR5-binding antibody after electrovaccination of BALB/c mice with pmDR5ectm-Td1 encoding a fusion protein of mouse DR5 and an immunogenic fragment of tetanus toxin. These findings support DR5 as a promising vaccine target for controlling TNBC and other DR5-positive cancers.