肿瘤坏死因子相关的凋亡诱导配体及其与肿瘤治疗前景

肿瘤坏因子相关的凋亡诱导配体（ＴＮＦ－related apoptosis inducing ligand,TRALL)属于肿瘤坏死因子家族,可激活肿瘤细胞的凋亡。本文介绍了ＴＲＡＩＬ的结构与功能,凋亡诱导途径及其肿瘤治疗应用前景。     

肿瘤坏死因子相关凋亡诱导配体研究进展

TRAIL(又称为Apo2L)是TNF超家族的新成员。它可以选择性诱导肿瘤细胞的凋亡,而对正常细胞无凋亡作用本介绍了TRAIL的结构和功能、凋亡途径、肝毒性研究及应用前景。TRAIL很可能成为新一代的抗肿瘤制剂。     

肿瘤坏死因子相关的凋亡诱导配体(TRAIL)基因抑制小鼠子宫基质细胞的蜕膜化

为了观察肿瘤坏死因子相关凋亡诱导配体(TRAIL)基因对体外培养的小鼠蜕膜基质细胞增殖及凋亡的作用,探讨TRAIL对小鼠子宫蜕膜化进程的影响,构建TRAIL过表达及干扰质粒,转染小鼠基质细胞后诱导蜕膜化发生.转染72h后,应用半定量RT-PCR和Western blotting检测蜕膜基质细胞中TRAILmRNA和蛋白质的表达情况、MTT法观察蜕膜基质细胞的生长和增殖能力、流式细胞术检测蜕膜基质细胞的细胞周期分布情况和凋亡率.经酶切和核苷酸测序证实,TRAIL基因正确克隆入真核表达载体且能够上调TRAIL的表达,干扰质粒能有效地抑制TRAIL基因的表达.TRAIL过表达和RNA干扰的结果表明:TRAIL具有将蜕膜基质细胞阻滞在G0/G1期、抑制蜕膜基质细胞增殖并促使其凋亡的功效,提示TRAIL可能参与调节胚胎植入后基质细胞的有序蜕膜化进程.     

肿瘤坏死因子家族新成员——TRAIL

肿瘤坏死因子相关的凋亡诱导配体(TRAIL)或称凋亡素2配体(Apo2 ligand, Apo-2L), 是TNF家族的新成员.它是从表达序列标签库(expressed sequenced tag, EST)中寻找TNF的同源分子时发现的.TRAIL是一种分子质量为32.5 ku的Ⅱ型跨膜糖蛋白, 活性形式呈同源三聚体.TRAIL和可溶性的TRAIL强烈诱导肿瘤细胞株凋亡.新近发现的TRAIL受体DR4和DR5及TRID说明了TRAIL与TNF和Fas/Apo-1配体的作用途径是不同的.随着对TRAIL的受体及作用机理研究的深入, TRAIL很可能成为新一代抗肿瘤制剂.     

胰腺癌对TNF相关凋亡诱导配体(TRAIL)的抗性与逆转研究

TRAIL(TNF-related apoptosis-inducing ligand)是一种能识别和选择性杀伤肿瘤细胞的蛋白质因子,但研究发现胰腺癌对TRAIL的敏感程度远远不及其他肿瘤,其抵抗机制主要集中于胞内水平的调节,如c-FLIPS、BCL-2/BCL-xL、XIAP表达上调等,且针对性的逆转策略也进行了深入的研究.本文就TRAIL途径在胰腺癌中的研究进展作一概要的介绍.     

乙醛脱氢酶1和肿瘤坏死因子相关诱导凋亡配体在膀胱癌组织中的表达及其临床意义

目的:探讨乙醛脱氢酶1(ALDH-1)和肿瘤坏死因子相关诱导凋亡配体(TRAIL)在膀胱癌组织中的表达及其临床意义。方法:选取2015年3月到2018年1月在河北北方学院附属第一医院进行治疗的膀胱癌患者70例,收集其手术切除的癌组织和癌旁正常组织,采用免疫组化法检测癌组织和癌旁正常组织中ALDH-1、TRAIL表达情况,分析ALDH-1、TRAIL的表达与膀胱癌患者的临床病理特征的关系及癌组织中ALDH-1、TRAIL表达的相关性。结果:癌组织中的ALDH-1的阳性表达率高于癌旁正常组织,TRAIL的阳性表达率低于癌旁正常组织(P0.05)。膀胱癌患者的ALDH-1阳性表达率与年龄、性别、分化程度、肿瘤数量无关(P0.05),临床分期为T2-T3期、有淋巴结转移的膀胱癌患者ALDH-1阳性表达率高于临床分期为Ta-T1期、无淋巴结转移的膀胱癌患者(P0.05)。膀胱癌患者的TRAIL阳性表达率与年龄、性别、临床分期、淋巴结转移、肿瘤数量无关(P0.05),高分化的膀胱癌患者TRAIL阳性表达率高于中低分化的膀胱癌患者(P0.05)。Pearson相关性分析显示,癌组织中ALDH-1、TRAIL表达无明显的相关性(P0.05)。结论:膀胱癌组织中ALDH-1的表达偏高且与临床分期和淋巴结转移有关,TRAIL的表达偏低且与分化程度有关,但ALDH-1和TRAIL之间无相关性,需进一步探讨与研究。     

TRAIL联合多西紫杉醇诱导喉鳞癌Hep-2 细胞凋亡的实验研究

目的：观察肿瘤坏死因子相关凋亡诱导配体( TRAIL) 联合多西紫杉醇应用于人喉鳞癌Hep-2 细胞生长的抑制增殖和诱导 凋亡作用。方法：实验分四组,1 组对照组,2 组为应用TRAIL组,3 组单独应用多西紫杉醇,4 组联合应用TRAIL及多西紫杉醇。 分别应用MTT、流式细胞仪检测细胞凋亡率,倒置显微镜观察细胞的形态学改变。结果：TRAIL 与多西紫杉醇联合作用于Hep-2 细胞,能显著增强对Hep-2 细胞的杀伤、抑制增殖及诱导凋亡作用,其联合应用的凋亡抑制率明显高于单独应用TRAIL组和多西 紫杉醇组( P<0.05)。结论：TRAIL与多西紫杉醇联用能显著提高对喉鳞癌Hep-2 细胞的生长抑制和诱导凋亡作用。     

TRAIL联合多西紫杉醇诱导喉鳞癌Hep-2细胞凋亡的实验研究

目的：观察肿瘤坏死因子相关凋亡诱导配体（TRAIL）联合多西紫杉醇应用于人喉鳞癌Hep-2细胞生长的抑制增殖和诱导凋亡作用。方法：实验分四组,1组对照组,2组为应用TRAIL组,3组单独应用多西紫杉醇,4组联合应用TRAIL及多西紫杉醇。分别应用MTT、流式细胞仪检测细胞凋亡率,倒置显微镜观察细胞的形态学改变。结果：TRAIL与多西紫杉醇联合作用于Hep-2细胞,能显著增强对Hep-2细胞的杀伤、抑制增殖及诱导凋亡作用,其联合应用的凋亡抑制率明显高于单独应用TRAIL组和多西紫杉醇组（P〈0．05）。结论：TRAIL与多西紫杉醇联用能显著提高对喉鳞癌Hep-2细胞的生长抑制和诱导凋亡作用。     

以TRAIL为靶点的肿瘤治疗研究进展

肿瘤坏死因子相关凋亡配体（tumor necrosis factor-related apoptosis-inducing ligand,TRAIL）是肿瘤坏死因子（tumor necrosis factor,TNF）超家族成员。TRAIL与其受体结合后启动凋亡信号转导,选择性地诱导肿瘤细胞凋亡,而对正常组织细胞没有明显的伤害,而且一些药物和细胞因子可协同TRAIL诱导肿瘤细胞凋亡。本文就TRAIL及其受体、TRAIL诱导凋亡的机制以及影响凋亡的因素和途径,以TRAIL为靶点的肿瘤治疗的研究现状作一综述。     

肿瘤坏死因子α和β对电离辐射诱导细胞凋亡的效应

为探讨肿瘤坏死因子（tumor necrosis foctor)α和β（TNFα和β）对电离辐射诱发细胞凋亡的效应及其机理,采用DNA琼脂糖凝胶电泳和FACS分析等方法,观察了人肿瘤坏死因子α（hTNFα)和β（hTNFβ)对^60Co-γ射线诱发细胞凋亡的形态学,生化学变化。结果显示：hTNFα或hTNFβ均可明显抑制^60Co-γ射线诱发正常人胚肺二倍体细胞（2BS）的凋亡,而相同剂量的hTNFα能促进^60Co-γ射线诱发的人体肺腺癌细胞系A549细胞凋亡,而对另一株人体肺癌SPC细胞的效应比A549降低1倍;hTNFβ能分别增强A549和SPC的细胞凋亡频率。由此认为,hTNFα和hTNFβ均可通过调节细胞的生理生化反应来改变细胞对电离辐射的敏感性,可保护正常细胞免受辐射损伤,而增加某些肿瘤细胞对辐射的敏感性。     

蛋氨酸脑啡肽联合白细胞介素-2抗肿瘤作用研究

观察蛋氨酸脑啡肽(MEK)和注射用白细胞介素-2(IL-2)单独和联合应用的抗肿瘤效应。应用动物移植性肿瘤的体内试验法,分别观察MEK、IL-2和(MEK+IL-2)对肿瘤的抑瘤作用及小鼠生命延长率情况。MEK组、IL-2组和(MEK+IL-2)组的抑瘤作用分别为131.38%(P0.01)、69.63%(P0.01)、229.170%(P0.01),都有显著差异;生命延长率为:MEK组18.27%(P0.01),有显著差异;IL-2组9.31%(P0.01),无显著差异;(MEK+IL-2)组32.07%(P0.01),有显著差异。结果表明:MEK、IL-2和(MEK+IL-2)都对小鼠移植性瘤有一定的抑制作用,MEK和(MEK+IL-2)能够延长小鼠生命,而IL-2单独应用不能延长小鼠生命;MEK和IL-2联合应用时作用相加,而不良反应未相加。     

Following a TRAIL： Update on a ligand and its five receptors

Identification of tumour necrosis factor apoptosis inducing ligand (TRAIL), a TNF family ligand, sparked a torrent of research, following an initial observation that it could kill tumour cells, but spare normal cells. Almost a decade after its discovery, and with five known receptors, the true physiological role of TRAIL is still debated and its anti-tumorigenic properties limited by potential toxicity. This review takes a comprehensive look at the story of this enigmatic ligand,addressing its remaining potential as a therapeutic and providing an overview of the TRAIL receptors themselves.     

TRAIL/TRAIL‐R in hematologic malignancies

Defects in apoptosis are observed in many cancer cell types and contribute in a relevant way to tumorigenesis. Apoptosis is a complex and well‐regulated cell death program that plays a key role in the control of cell homeostasis, particularly at the level of the hematopoietic system. Apoptosis can be initiated through two different mechanisms involving either activation of the death receptors (extrinsic pathway) or activation of a mitochondrial apoptotic process (intrinsic pathway). Among the various death receptors a peculiar role is played by TNF‐related apoptosis‐inducing ligand (TRAIL)‐receptors (TRAIL‐Rs) and their ligand TRAIL. TRAIL recently received considerable interest for its potent anti‐tumor killing activity, sparing normal cells. Here, we will review the expression and the abnormalities of TRAIL/TRAIL‐R system in hematologic malignancies. The large majority of primary hematologic tumors are resistant to TRAIL‐mediated apoptosis, basically due to the activation of anti‐apoptotic signaling pathway (such as NF‐κB), overexpression of anti‐apoptotic proteins (such as FLIP, Bcl‐2, XIAP) or expression of TRAIL decoy receptors or reduced TRAIL‐R1/‐R2 expression. Strategies have been developed to bypass this TRAIL resistance and are based on the combination of TRAIL with chemotherapy or radiotherapy, or with proteasome or histone deacetylase or NF‐κB inhibitors. The agents used in combination with TRAIL either enhance TRAIL‐R1/‐R2 expression or decrease expression of anti‐apoptotic proteins (c‐FLIP, XIAP, Bcl‐2). Many of these combinatorial therapies hold promise for future developments in treatment of hematologic malignancies. J. Cell. Biochem. 110: 21–34, 2010. © 2010 Wiley‐Liss, Inc.     

抗体靶向超抗原的抗肿瘤作用研究进展

作为一种强大的T细胞激活剂,超抗原具有激活T细胞杀伤HLA-DR 肿瘤细胞的能力,但这种杀伤作用无特异性。抗体靶向超抗原通过抗肿瘤抗体与超抗原偶联,既具有超抗原活性,又具有肿瘤靶向性,从而可以选择性地结合到目标细胞,进行有效的特异性杀伤,因此,抗体靶向超抗原在肿瘤的生物治疗领域具有广泛的应用前景。     

PHA-767491联合TRAIL抑制肝癌细胞Bel-7404增殖

将小分子抗癌药物PHA-767491和携带TRAIL基因的非复制型腺病毒(Ad-TRAIL)共同作用于肝癌细胞Bel-7404,探讨PHA-767491联合TRAIL蛋白对肝癌细胞增殖的协同抑制作用及其作用机理. MTT法检测细胞存活率,Hoechst 33342荧光检测细胞凋亡现象和流式细胞仪检测细胞凋亡水平. 结果表明,PHA-767491联合Ad-TRAIL抑制Bel-7404细胞增殖的能力显著优于单一用药. Western印迹进一步分析蛋白表达水平显示,PHA-767491可以通过下调抗凋亡蛋白Mcl-1和Xiap的表达,从而显著增强TRAIL蛋白诱导Bel-7404细胞凋亡的能力;PHA-767491联合Ad-TRAIL处理Bel-7404细胞后,不仅Bel-7404细胞凋亡水平显著增加,并且伴随着PARP和Caspase3的大量剪切. 本研究证实, PHA-767491和TRAIL的联合使用对抑制肝癌细胞Bel-7404增殖表现出了显著的协同效应,为今后癌症药物的联合治疗提供了新的思路.     

TNF-related apoptosis inducing ligand (TRAIL) and its receptors in tumor surveillance and cancer therapy

TNF-related apoptosis-inducing ligand (TRAIL/APO-2L) is a typical member of the TNF ligand family that induces apoptosis by activating the death receptors TRAIL-R1 and TRAIL-R2. TRAIL has attracted great attention in recent years as a promising anti cancer reagent because recombinant soluble TRAIL derivatives induce apoptosis in a broad range of tumor cells but not or only rarely in non-transformed cells. In this review we will address the putative role of TRAIL in cancer treatment in the light of the emerging importance of TRAIL in tumor surveillance and discuss the molecular basis of the cooperation of TRAIL and chemotherapeutic drugs. In particular, we debate controversial data in the literature concerning the cytotoxicity of different TRAIL derivatives on primary human cells.     

肿瘤坏死因子相关的凋亡诱导配体(TRAIL)cDNA的克隆、表达和活性测定

肿瘤坏死因子相关的凋亡诱导配体 (TRAIL)能选择性诱导肿瘤细胞凋亡 .为利用基因工程技术获得重组TRAIL蛋白可溶性片段 (sTRAIL) ,设计 1对引物 .利用PCR技术特异性扩增出sTRAIL的cDNA ,克隆于质粒pGEM 3Zf( )的EcoRⅠ和PstⅠ位点 .经测序证明序列正确后克隆于表达质粒pBV2 2 0的EcoRⅠ和PstⅠ位点 ,转化大肠杆菌DH5α .转化菌株经温度诱导 ,SDS PAGE检测和Western印迹鉴定 ,获得重组sTRAIL的高水平非融合表达菌株 .表达量占菌体总蛋白的 2 0 % .对其表达产物进行了初步纯化 ,SDS PAGE结果显示纯度可达 90 %以上 .用L92 9细胞测定其生物学活性表明 ,重组蛋白在体外能明显诱导肿瘤细胞凋亡     

TRAIL apoptosis is enhanced by quercetin through Akt dephosphorylation

TNF-related apoptosis-inducing ligand (TRAIL) is a promising cancer therapy that preferentially induces apoptosis in cancer cells. However, many neoplasms are resistant to TRAIL by mechanisms that are poorly understood. Here we demonstrated that human prostate cancer cells, but not normal prostate cells, are dramatically sensitized to TRAIL-induced apoptosis and caspase activation by quercetin. Quercetin, a ubiquitous bioactive plant flavonoid, has been shown to inhibit the proliferation of cancer cells. We have shown that quercetin can potentiate TRAIL-induced apoptotic death. Human prostate adenocarcinoma DU-145 and LNCaP cells were treated with various concentrations of TRAIL (10-200 ng/ml) and/or quercetin (10-200 microM) for 4 h. Quercetin, which caused no cytotoxicity by itself, promoted TRAIL-induced apoptosis. The TRAIL-mediated activation of caspase, and PARP (poly(ADP-ribose) polymerase) cleavage were both enhanced by quercetin. Western blot analysis showed that combined treatment with TRAIL and quercetin did not change the levels of TRAIL receptors (death receptors DR4 and DR5, and DcR2 (decoy receptor 2)) or anti-apoptotic proteins (FLICE-inhibitory protein (FLIP), inhibitor of apoptosis (IAP), and Bcl-2). However, quercetin promoted the dephosphorylation of Akt. Quercetin-induced potent inhibition of Akt phosphorylation. Taken together, the present studies suggest that quercetin enhances TRAIL-induced cytotoxicity by activating caspases and inhibiting phosphorylation of Akt.     

Rapid and efficient cancer cell killing mediated by high-affinity death receptor homotrimerizing TRAIL variants

The tumour necrosis factor family member TNF-related apoptosis-inducing ligand (TRAIL) selectively induces apoptosis in a variety of cancer cells through the activation of death receptors 4 (DR4) and 5 (DR5) and is considered a promising anticancer therapeutic agent. As apoptosis seems to occur primarily via only one of the two death receptors in many cancer cells, the introduction of DR selectivity is thought to create more potent TRAIL agonists with superior therapeutic properties. By use of a computer-aided structure-based design followed by rational combination of mutations, we obtained variants that signal exclusively via DR4. Besides an enhanced selectivity, these TRAIL-DR4 agonists show superior affinity to DR4, and a high apoptosis-inducing activity against several TRAIL-sensitive and -resistant cancer cell lines in vitro. Intriguingly, combined treatment of the DR4-selective variant and a DR5-selective TRAIL variant in cancer cell lines signalling by both death receptors leads to a significant increase in activity when compared with wild-type rhTRAIL or each single rhTRAIL variant. Our results suggest that TRAIL induced apoptosis via high-affinity and rapid-selective homotrimerization of each DR represent an important step towards an efficient cancer treatment.