TRAIL、TRAIL-R在原发性肝癌及其癌旁组织中的表达

目的 检测肿瘤坏死因子相关凋亡诱导配体（TRAIL）及其受体在原发性肝癌（PHC）及癌旁组织中的表达。方法 收集PHC及癌旁组织各30例,采用半定量RT-PCR方法,对TRAIL及其受体的表达作半定量检测。结果 （1）30例痈组织度癌旁组织中均有TRAIL及其死亡受体4（DR4）、死亡受体5（DR5）、“诱骗”受体1,2（DcR1、DeR2）的表达。（2）TRAIL、DcR1及DcR2在癌组织中的表达量较之癌旁组织明显降低（P〈0．01）,而DR4、DR5的表达量则高于癌旁组织;（3）PHC组织中4种受体之间,DR4及DR5的表达量明显高于DcR1及DcR2,且PHC组织中4种受体间的表达量存在相关性。结论 PHC及其癌旁组织中均存在TRAIL及其受体的表达,且有表达量的差异。     

肿瘤坏死因子相关凋亡诱导配体诱导人脑胶质瘤细胞凋亡作用的研究

目的研究肿瘤坏死因子相关凋亡诱导配体（TRAIL）受体在不同级别胶质瘤中的表达及TRAIL对原代培养的胶质瘤细胞凋亡的诱导作用。方法应用原位杂交的方法检测TRAIL受体在45例胶质瘤和5例正常脑组织中的转录表达；分别对其中22例和3例进行原代培养，用流式细胞仪检测TRAIL作用后细胞的凋亡率。结果在45例胶质瘤中，DR4、DR5、DcR1、DcR2的mRNA的表达例数分别为38例、41例、40例、2例。成功培养出18例胶质瘤细胞和3例正常胶质细胞，TRAIL作用后凋亡率在80％以上、50％～80％、50％以下的例数分别为5例、9例、4例，正常胶质细胞无明显凋亡发生。结论TRAIL受体的表达与肿瘤的恶性程度无关；TRAIL能有效地选择性杀死胶质瘤细胞。     

FasL、TRAIL及其受体DcR1在宫颈癌中的表达

目的：探讨FasL、TRAIL及其受体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FasL、TRAIL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综合分析。结果：FasL在正常宫颈、CIN和宫颈癌组织中的表达呈递增趋势;宫颈癌组织中,FasL阳性表达随组织学分级的降低而升高;有淋巴结转移者FasL阳性表达明显高于无淋巴结转移者;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。结论：FasL．TRAIL及DcR1的异常表达在宫颈癌变过程中起到了一定的作用。     

TRAIL及其受体在咖啡因抑制肝癌细胞系HepG2增殖中的作用

目的探讨TRAIL及其受体在咖啡因(caffeine)抑制肝癌细胞系HepG2增殖中的作用。方法HepG2细胞分别经caffeine、TRAIL及caffeine+TRAIL作用24h,采用MTT法检测HepG2细胞增殖抑制情况,根据中效原理进行联合用药效应评价;流式细胞仪检测细胞凋亡和细胞周期分布;Westernblot法检测caffeine作用不同时间HepG2细胞中TRAIL受体相关蛋白的表达。结果在1.25～20mmol.L-1浓度范围内,caffeine明显抑制HepG2细胞增殖;在0.01275～0.2040μmol.L-1浓度范围内,TRAIL可明显抑制HepG2细胞增殖。Caffeine联合TRAIL在多数效应范围内的合用指数小于1,具有协同作用。Caffeine5mmol.L-1和TRAIL0.0510μmol.L-1联合用药组HepG2细胞凋亡率明显高于各单独用药组,且两者联合用药对HepG2细胞周期具有明显的影响,使G0/G1期细胞比例明显增加,S期及G2/M期细胞比例明显减少;caffeine5mmol.L-1作用HepG2细胞24h时,其DR4及DR5的表达量明显增加,而DcR1和DcR2的表达无改变。结论TRAIL在caffeine抑制HepG2细胞增殖过程中具有一定的协同作用,其机制可能与caffeine上调HepG2细胞表面DR4、DR5的表达,联合TRAIL后能够进一步诱导凋亡及调节细胞周期有关。     

5-氮杂胞苷对胃癌细胞系的生长凋亡及DR4与DR5表达的影响

目的探讨5-氮杂胞苷(5-Aza-CdR)对胃癌细胞系的生长,凋亡及DR4与DR5表达的影响。方法反转录聚合酶链反应(RT-PCR)方法、Western blot方法和甲基化特异性PCR(MS-PCR)法检测5-Aza-CdR处理后的胃癌细胞株内DR4和DR5的RNA表达水平。结果 5-Aza-CdR能够逆转胃癌细胞中DR4和DR5的启动子甲基化水平,并上调DR4和DR5的表达水平。结论 5-氮杂胞苷对胃癌细胞系MKN28、AGS和SGC7901具有增殖抑制作用,且具有药物浓度和作用时间的依赖性。促进胃癌细胞系MKN28、AGS和SGC7901的凋亡且能使死亡受体DR4和DR5的表达水平增高。     

早孕绒毛TRAIL及其受体在自然流产中的作用机制研究

目的 通过检测自然流产患者和正常早期妊娠妇女的绒毛中肿瘤坏死因子相关凋亡诱导配体(TNF-related apoptosis inducing ligand,TRAIL)及其受体DR5(death receptor-5)、DcRl(decoy receptor 1)的表达,来探讨肿瘤坏死因子相关凋亡诱导配体(TRAIL)及其受体DR5、DcRl在自然流产发病过程中的作用机制.方法 收集自然流产患者30例,正常早期妊娠妇女30例,使用半定量RT-PCR的方法检测自然流产者和正常早期妊娠者绒毛组织中TRAIL、DR5和DcRl基因mRNA的转录水平,同时用免疫组化的方法检测两组绒毛中TRAIL、DR5和DcRl蛋白的表达情况.结果 自然流产组绒毛组织中TRAIL、DR5基因mRNA的转录水平明显高于正常妊娠组(P<0.01),而DcRl基因mRNA的转录水平明显低于正常妊娠组(P<0.01).自然流产组绒毛组织中TRAIL、DR5蛋白的表达明显高于正常妊娠组(P<0.01),而绒毛组织中DcRl蛋白的表达很少或无,自然流产组中DcRl蛋白表达的水平明显低于正常妊娠组(P<0.01).结论 绒毛组织中肿瘤坏死因子相关凋亡诱导配体(TRAIL)及其受体(DR5、DcRl)的表达异常,从而可以导致细胞凋亡的异常,可能是导致自然流产的发病原因之一.     

DR在胸部体检中的应用

目的:评价DR(直接数字化X线摄影系统)在胸部体检中的应用价值。资料与方法:收集胸部体检1132例,均用DR进行胸部摄片,分析其中阳性病例,寻找DR检查的优势。结果:1132例体检者中发现阳性患者134例,其中肺结核38例(占体检人数的3.35%),慢支肺气肿23例(2.03%),肺癌1例(0.08%),肺结节37例(3.26%),不同程度心影增大35例(3.09%)。结论:DR在胸部体检中具有广泛的应用价值。     

线粒体通路和死亡受体通路在中华眼镜蛇毒组分诱导KG1a细胞凋亡中的作用

目的研究线粒体通路和死亡受体通路在中华眼镜蛇毒组分(naja naja actra venom component,NNAVC)诱导KG1a细胞凋亡过程中的作用。方法以人急性髓系白血病细胞株KG1a细胞为研究对象,采用Annexin V-FITC/PI荧光染色法进行凋亡细胞的形态学观察,流式细胞仪检测细胞凋亡率,分光光度法检测Caspase-9、Caspase-8和Caspase-3的活性,ELISA法检测胞质内细胞色素C的表达水平,流式细胞仪分析TRAIL死亡受体(DR4、DR5)和诱骗受体(DcR1、DcR2)的改变。结果 NNAVC作用后,荧光显微镜下可见明显的凋亡细胞,细胞凋亡率随药物作用浓度的增加而升高;Caspase-9、Caspase-8和Caspase-3蛋白被激活,胞质内细胞色素C的表达明显增加;流式细胞仪检测结果显示,NNAVC具有降低死亡受体DR4和诱骗受体DcR1的表达率,而提高DR5和DcR2的表达水平的作用。结论线粒体凋亡通路和死亡受体通路均参与NNAVC诱导KG1a细胞凋亡的过程,这可能是NNAVC发挥抗白血病作用的机制之一。     

1516例HLA-A、B、DR分型实验体会

我院自 1996年 12月起开展人白细胞抗原 (HL A- A、 B、DR)分型技术 ,至今已检测标本 15 16例 ,并在肾脏、骨髓、肝脏、心脏、肺脏、角膜等器官和组织移植领域广泛应用。分析总结 15 16例不同病种的 HL A- A、 B、 DR分型方法及技巧 ,以便于实验操作标准化、规范化 ,达到分型结果准确可靠。1　临床资料收集行 HL A- A、 B、 DR分型的全血、睥脏、脐血样本共15 16份 ,其中应用于肾移植 76 9例 ,骨髓移植 2 0例 ,心脏移植 5例 ,肝脏移植 17例 ,肺脏移植为 1例 ,胰肾联合移植 2例 ,角膜移植 14例 ,其它 6 88例为各种移植供者和科研样…     

TRAIL、DR5和caspase3在颞下颌关节紊乱病关节盘新生血管细胞中的表达

目的:观察颞下颌关节紊乱病关节盘新生血管中TRAIL、DR5、caspase-3的表达程度,评估新生血管细胞的自限性凋亡过程。方法:对取自15名颞下颌关节盘移位患者以及4个正常关节盘进行TRAIL,DR5及caspase-3的免疫组织化学研究。结果:研究证实在新生血管内皮及中间层可高免疫活性表达TRAIL、DR5以及caspase-3。结论:在颞下颌关节退行性变中伴有局部血管生成,并可激活血管上皮细胞凋亡过程而表现自限性的血管退化;新生血管的自我抑制,在切断肿瘤血供或减少炎症损伤中可起到重要作用。     

TRAIL、MMP-2及TIMP-2在大肠癌中的表达及意义的研究

目的研究肿瘤坏死因子相关凋亡诱导配体（TRAIL）、MMP-2、TIMP-2在大肠癌中的表达及意义,探讨TRAIL诱导肿瘤细胞凋亡的可能机制,MMP-2及TIMP-2在肿瘤的侵袭和转移中的重要的作用及相互关系,寻求提高TRAIL对肿瘤细胞敏感性的可能途径。方法采用SP法检测102例大肠癌及其癌旁5cm组织、25例正常大肠黏膜组织中TRAIL、MMP-2及TIMP-2蛋白的表达水平。结果 TRAIL在大肠癌、癌旁组织及正常大肠黏膜组织中的表达呈明显递增趋势（P〈0.01）;MMP-2在大肠癌、癌旁组织及正常大肠黏膜组织中的表达呈明显递减趋势（P〈0.01）;TIMP-2在大肠癌、癌旁组织及正常大肠黏膜组织中的表达无统计学意义（P〉0.05）。TRAIL在中、低分化癌中的表达明显低于高分化癌中的表达（P=0.003）。结论 TRAIL、MMP-2及TIMP-2可能与大肠癌的发生、发展、侵袭及转移密切相关;MMP-2与TIMP-2在癌组织中表达失衡在肿瘤的侵袭及转移中可能起重要作用。通过改善MMP-2与MMP-2在癌组织中表达的失衡,可能提高TRAIL对肿瘤细胞的特异性杀伤效率及改善肿瘤患者的预后。     

P-glycoprotein enhances TRAIL-triggered apoptosis in multidrug resistant cancer cells by interacting with the death receptor DR5

The death-inducing cytokine TRAIL is a promising agent for anticancer therapy since it preferentially kills cancer versus normal cells; however, some cancer cells are TRAIL-resistant. We initially explored whether overexpression of the MDR1 gene product P-glycoprotein (P-gp), which causes multidrug resistance (MDR) in cancer cells, also contributes to TRAIL-resistance. Surprisingly, our results revealed that P-gp-overexpression enhances TRAIL-induced apoptosis not only in neoplastic cells transfected with the MDR1 gene but also in MDR variants selected with cytotoxic anticancer agents. Mechanistic analysis of TRAIL-induced apoptosis in the MDR1-transfected MCF-7 breast cancer cell line BC-19 revealed that TRAIL-triggered significantly more apoptosis in these cells compared with parental MCF-7 cells by binding to the TRAIL receptor DR5. DR5 but not DR4 engagement by TRAIL attenuated cellular ATP levels by robustly stimulating P-gp ATPase activity, and thus triggered P-gp-dependent apoptosis by depletion of the cellular ATP pool. In addition to hyperactive P-gp-mediated ATP hydrolysis, TRAIL-induced, P-gp-potentiated apoptosis was associated with activation of caspases-6, -7, -8, and -9; Bid cleavage; and mitochondrial depolarization. P-gp interacted with the TRAIL receptors DR4, DR5, and DcR1 in plasma membranes and enhanced TRAIL binding to DR5. Interestingly, the decreased level of the decoy TRAIL receptor, DcR1, in BC-19 cells further sensitized these cells to TRAIL. Therefore, both extrinsic and intrinsic apoptosis pathways are involved in this process. These findings for the first time reveal that TRAIL treatment preferentially causes apoptosis in P-gp-overexpressing MDR cells, and suggests significant clinical implications for the use of TRAIL in treating neoplasms that have failed chemotherapy.     

Capsaicin sensitizes TRAIL-induced apoptosis through Sp1-mediated DR5 up-regulation: involvement of Ca(2+) influx

Although tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induces apoptosis in various malignant cells, several cancers including human hepatocellular carcinoma (HCC) exhibit potent resistance to TRAIL-induced cell death. The aim of this study is to evaluate the anti-cancer potential of capsaicin in TRAIL-induced cancer cell death. As indicated by assays that measure phosphatidylserine exposure, mitochondrial activity and activation of caspases, capsaicin potentiated TRAIL-resistant cells to lead to cell death. In addition, we found that capsaicin induces the cell surface expression of TRAIL receptor DR5, but not DR4 through the activation Sp1 on its promoter region. Furthermore, we investigated that capsaicin-induced DR5 expression and apoptosis are inhibited by calcium chelator or inhibitors for calmodulin-dependent protein kinase. Taken together, our data suggest that capsaicin sensitizes TRAIL-mediated HCC cell apoptosis by DR5 up-regulation via calcium influx-dependent Sp1 activation.     

Inostamycin enhanced TRAIL-induced apoptosis through DR5 upregulation on the cell surface

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has been considered as a possible therapeutic agent for cancer treatment. This is because of its selective cytotoxicity against various cancer cells without a detrimental effect on normal cells. However, recent studies have reported that the potential application of TRAIL in cancer therapy is limited, as many cancer cells have been found to be resistant to TRAIL. Therefore, small molecule compounds that potentiate the cytotoxicity of TRAIL would be strategic candidates for therapeutic applications in combination with TRAIL. Here we found that a combined treatment of inostamycin and TRAIL synergistically induced caspase-dependent apoptosis in HCT116 cells. Inostamycin upregulated DR5, and a knockdown of DR5 suppressed the apoptosis that was synergistically induced by co-treatment with inostamycin and TRAIL. Moreover, inostamycin increased the expression of DR5 on the cell surface. Therefore, inostamycin-increased cell surface expression of DR5 may have contributed to the enhancement of TRAIL-induced apoptosis. Our study suggests that combined treatment with inostamycin and TRAIL may offer a strategy to overcome TRAIL resistance in tumor cells.