肿瘤坏死因子凋亡诱导配体诱导膀胱癌EJ细胞凋亡的研究

肿瘤坏死因子相关凋亡诱导配体(TRAIL)是能够较特异性诱导肿瘤细胞凋亡的肿瘤坏死因子超家族成员 ,由于对正常组织不表现毒性[1] ,在肿瘤治疗方面得到广泛研究。我们对TRAIL诱导膀胱癌细胞EJ凋亡的作用进行了研究 ,现报告如下。材料与方法　重组人TRAIL(rhTRAIL)由KamiyaBiomedical公司制备和纯化 ,浓度设置为 10 0、5 0、2 0和 10ng/ml。Annexin V FluosStainingKit试剂盒购自罗氏公司。人膀胱癌EJ细胞用含 10 %小牛血清 ,10 0U/ml青霉素和10 0 μg/ml链霉素的RPMI 16 4 0 37℃、5 0ml/LCO2 、饱和湿度下培养。细胞按 5× …     

转染肿瘤坏死因子相关凋亡诱导配体对膀胱癌细胞的抑制作用

目的探讨肿瘤坏死因子相关凋亡诱导配体（TRAIL）对膀胱癌细胞的抑制作用。方法构建TRAIL融合增强绿色荧光蛋白（EGFP）真核表达载体，脂质体法转染膀胱癌细胞株玎细胞。荧光显微镜下计算转染率。分别采用逆转录-聚合酶链反应（RT-PCR）、Western blot检测TRAIL的表达。采用流式细胞仪检测细胞凋亡，噻唑蓝（MTT）法、细胞倍增时间和集落形成率观察TRAIL的抑制作用。结果48h观察重组载体转染率为38．4％，空载体转染率为35．3％（P〉0．05）．RT-PCR和Western blot证实转染后EJ细胞中TRAIL的表达明显增加；凋亡率明显高于转染空白载体和未转染组（P〈0．01）；EJ细胞增殖受到明显抑制（P〈0．01）。结论TRAIL具有诱导膀胱癌细胞凋亡、抑制增殖的作用，有望成为治疗膀胱癌的新方法。     

肿瘤坏死因子相关凋亡诱导配体与胰腺癌及相关药物研究

自肿瘤坏死因子家族的新成员肿瘤坏死因子相关凋亡诱导配体被发现以来,它一直是近些年来在肿瘤研究方面的热点.本文综述了近些年肿瘤坏死因子相关凋亡诱导配体在胰腺癌方面的相关研究进展,从肿瘤坏死因子相关凋亡诱导配体诱导胰腺癌肿瘤细胞凋亡机制人手,简要介绍胰腺癌对肿瘤坏死因子相关凋亡诱导配体的耐药机制.并根据目前已认同的耐药机制...     

肿瘤坏死因子相关凋亡诱导配体治疗肿瘤存在的问题及策略

肿瘤坏死因子相关凋亡诱导配体(TRAIL)是近几年发现的TNF超家族成员 ,为Ⅱ型跨膜蛋白 ,能与其受体(TRAILR)结合 ,启动细胞内的信号转导 ,激活Caspase级联反应诱导细胞凋亡。这种新发现的凋亡诱导途径有重要的生物学意义 ,尤其是在抗肿瘤方面具有选择性杀伤肿瘤细胞同时赦免正常细胞的作用 ,因而引起了人们的高度重视 ,认为有望找到一个全新的肿瘤治疗方法。然而 ,随着对TRAIL研究的深入 ,TRAIL治疗肿瘤可能存在的障碍不断出现 ,人们对TRAIL能否成为一个理想的抗癌新药提出质疑 ,现从TRAIL治疗肿瘤可能存在的问题作一综述 ,并…     

肿瘤坏死因子相关凋亡诱导配体及其受体与抗肿瘤治疗

肿瘤坏死因子相关凋亡诱导配体(ｔｕｍｏｒｎｅｃｒｏｓｉｓｆａｃｔｏｒｒｅｌａｔｅｄａｐｏｐｔｏｓｉｓ ｉｎｄｕｃｉｎｇｌｉｇａｎｄ ,ＴＲＡＩＬ)是近几年发现的肿瘤坏死因子 (ｔｕｍｏｒｎｅｃｒｏｓｉｓｆａｃｔｏｒ,ＴＮＦ)超家族成员 ,为Ⅱ型跨膜蛋白 ,能与其受体 (ｔｕｍｏｒｎｅｃｒｏｓｉｓｆａｃｔｏｒｒｅｌａｔｅｄａｐｏｐｔｏｓｉｓ ｉｎｄｕｃｉｎｇｌｉｇａｎｄｒｅｃｅｐｔｏｒ,ＴＲＡＩＬＲ)结合 ,启动细胞内的信号转导 ,激活半胱—天冬氨酸蛋白酶 (ｃｙｓｔｅｉｎｅｃｏｎｔａｉｎｉｎｇａｓｐａｒａｔｅｓｐｅｃｉ…     

肿瘤坏死因子凋亡诱导配体诱导膀胱癌EJ细胞凋亡的研究

目的 探讨肿瘤坏死因子凋亡诱导配体(TRAIL)诱导膀胱癌细胞凋亡的效应。方法 10～100μg／L梯度浓度的TRAIL与培养的EJ细胞共孵育4～24h。采用Annexin V染色，DNA云梯实验和流式细胞技术检测诱导后EJ细胞凋亡发生的时期和程度。结果 TRAIL诱导4h即后可观察到早期细胞凋亡发生，8h后即可检测DNA片段化，24h后凋亡细胞最多达到66．29％。结论 TRAIL能够迅速诱导EJ细胞凋亡，可能成为膀胱癌治疗的新方法。     

肿瘤坏死因子相关的凋亡诱导配体在前列腺癌治疗中的研究进展

肿瘤坏死因子相关的凋亡诱导配体(TRAIL)是近年来研究发现的肿瘤坏死因子超家族新成员,其广泛存在于人体组织中,参与机体免疫调节、免疫自稳和免疫监视。TRAIL受体表达于多种细胞表面。研究表明,TRAIL能够诱导肿瘤细胞凋亡,对正常细胞无明显不不良反应,TRAIL良好的抗肿瘤活性和安全性得到了广泛的认识。前列腺癌的发生发展受细胞凋亡机制调控,TRAIL能诱导前列腺癌细胞凋亡,在前列腺癌的治疗中具有广阔前景。     

肿瘤坏死因子相关凋亡诱导配体介导脓毒症免疫抑制的研究进展

背景 肿瘤坏死因子相关凋亡诱导配体(tumor necrosis factor related apoptosis induced ligand,TRAIL)是肿瘤坏死因子超家族成员之一,其与受体DR4/DR5结合可以诱导肿瘤细胞的凋亡,而备受关注. 目的 总结TRAIL通路在脓毒症中的可能机制,探讨其在脓毒症疾病过程中的调控作用. 内容 脓毒症动物模型的研究发现,TRAIL可以加快活化的中性粒细胞发生凋亡并参与脓毒症免疫耐受的形成. 趋势 TRAIL受体在肿瘤、自身免疫性疾病、感染、心血管疾病、器官移植等领域的研究越来越多,作用机制也在不断地阐明.TRAIL及其受体在脓毒症疾病过程中发挥重要调节作用.     

肿瘤坏死因子相关凋亡诱导配体及其受体在肝癌组织中的表达的研究

肿瘤坏死因子相关凋亡诱导配体 (tumornecrosisfactorrelatedapopto sis inducingligand,TRAIL)是近来发 现的TNF超家族成员,为II型跨膜蛋 白,与其受体(TRAILR)结合并启动细 胞内的信号传导,在体外强烈诱导多种 肿瘤细胞凋亡,而正常细胞对其不敏 感,TRAIL的这一特性引起了学者的 高度重视,认为有望找到一种新的肿瘤 治疗方法。本文通过研究TRAIL及其 受体在肝细胞癌(HCC)中的表达,探讨 TRAIL用于治疗HCC的潜在可能性。 1.材料和方法:(1)肝癌组织:50 例肝细胞肝癌组织选自同济医院肝脏 外科中心手术切除标本,全部标本…     

肿瘤坏死因子相关诱导配体受体在前列腺癌组织中的表达

目的：研究肿瘤坏死因子相关诱导配体受体在前列腺癌组织中的表达情况。方法：应用RT—PCR检测20例良性前列腺增生和20例前列腺癌组织中的死亡受体DR4、DR5、假受体DcRl、DcR2的mRNA表达。结果：良性前列腺增生组织中DR4、DR5、DcR1均为85％(17／20)；DcR2为75％(15／20)。前列腺癌组织中死亡受体DR41、DR5为80％(16／20)；DcR1阳性表达率为15％(3／20)；假受体DcR2于前列腺癌组织中未见表达。结论：前列腺癌组织中的DcR1、DcR2受体缺乏表达，DcR1、DcR2在前列腺癌的凋亡调控途径中可能有重要作用。     

肿瘤坏死因子相关凋亡诱导配体联合化疗药物杀伤胰腺癌细胞的研究

目的研究肿瘤坏死因子相关凋亡诱导配体以及联合应用亚毒性剂量的化疗药物对胰腺癌的治疗作用。方法半定量 RT-PCR 检测 TRAILR mRNA 在胰腺癌细胞株 Canpan-2中的表达。DNA 琼脂糖凝胶电泳检测细胞凋亡情况。应用不同浓度的 TRAIL 及联合亚毒性剂量的氟尿嘧啶(5-fluorouracil)、吉西他滨(gemcitabin)处理胰腺癌细胞,通过 MTT 法检测细胞毒性作用,流式细胞仪分析细胞凋亡率。结果死亡受体 DR4、DR5及诱骗受体 DcR1、DcR2在胰腺癌细胞株 Canpan-2中均有表达。DNA 琼脂糖凝胶电泳可见到典型的凋亡梯形带。TRAIL100 ng/ml 作用胰腺癌细胞24h 后,细胞杀伤率为(29.5±1.2)%,且其作用存在浓度依赖性;联合应用亚毒性剂量的氟尿嘧啶、吉西化滨能够大大提高TRAIL 的细胞毒活性,杀伤率分别为 TRAIL+氟尿嘧啶(43.7±1.4)%,TRAIL+吉西化滨(49.8±1.2)%,联合用药前后差异有显著性(P<0.01)。结论 TRAIL 受体在胰腺癌细胞普遍表达,并存在受体类型的表达差异;TRAIL 与化疗药物氟尿嘧啶、吉西他滨有协同杀伤胰腺癌细胞的作用。     

Comparison of the cytotoxic activities of chemotherapeutic drugs using a human bladder cancer cell line

Summary Many chemotherapeutic drugs have been used to treat patients with advanced bladder cancer, but few of these have been evaluated adequately in phase II clinical trials. Continuous cell lines provide one means for comparing the in vitro cytotoxicities of anticancer agents. In this study, a continuous cell line derived from a transitional cell cancer of the human bladder, which still produces tumours histologically similar to the tumour of origin on xenotransplantation, was used to measure the in vitro cytotoxicities of twelve chemotherapeutic drugs by clonogenic assay. The most cytotoxic agents tested were methotrexate, mitoxantrone, adriamycin, mitomycin C and cisplatin. These in vitro findings are compatible with the activity of these drugs given systemically as single agents in phase II clinical trials in patients with advanced bladder cancer.     

干扰素结合化疗药物对膀胱癌细胞株BIU-87的抑制作用

在离体条件下，采用噻唑蓝（MTT）染色法，观察干扰素（IFN）与丝裂霉素C（MMC）、阿霉素（ADM）、噻替哌（TSPA）和羟基喜树碱（HCPT）等化疗药物联合应用对人类膀胱癌细胞株BIU－87的抑制作用。结果在连续联合用药组，浓度为25000u／ml的IFN与上述药物50％抑制率（ID50－1）剂量联合使用的抑制率分别为57．4％、58．6％、55．3％和64．3％；在序贯联合用药组，将MMC、ADM的ID50－2剂量作用于细胞24h后，再使用浓度为25000u／ml的IFN，24h后两者联合使用的抑制率分别为59．4％和54．3％。认为IFN与MMC等化疗药物联合应用能够取得较好的协同作用，为临床治疗浅表性膀胱肿瘤提供了新的思路。     

Neoadjuvant and adjuvant chemotherapy in muscle-invasive bladder cancer

Context

The use of neoadjuvant and adjuvant chemotherapy in the treatment of muscle-invasive bladder cancer is still controversial.

Objective

To determine the optimal use of chemotherapy in the neoadjuvant and adjuvant settings in patients with advanced urothelial cell carcinoma. Bladder preservation is also discussed.

Evidence acquisition

A critical review of the published literature on chemotherapy for patients with locally advanced bladder cancer was performed.

Evidence synthesis

The presence of occult micrometastases at the time of radical cystectomy leads to both distant and local failure in patients with locally advanced transitional cell carcinoma of the bladder. Both neoadjuvant and adjuvant therapies have been evaluated in patients with locally advanced bladder cancer. Studies evaluating adjuvant chemotherapy have been limited by inadequate statistical power to detect meaningful clinical answers as well as by experimental arms utilizing inadequate chemotherapy.

Conclusions

The aggregate of available evidence suggests that neoadjuvant cisplatin-based combination chemotherapy should be considered as a standard of care for patients with muscle-invasive or locally advanced operable bladder cancer. In patients who are either unfit for or refuse radical cystectomy, neoadjuvant chemotherapy with or without radiation can render bladder preservation possible for patients who attain an excellent clinical response. With the introduction of new cytotoxic drugs, there is a need for well-designed studies to address the optimal utility of perioperative therapy in high-risk patients with bladder cancer.     

低剂量阿霉素联合TRAIL诱导人前列腺癌细胞LNCAP凋亡的实验研究

目的 研究低剂量阿霉素联合肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导人前列腺癌细胞LNCAP凋亡的作用和机制,为提高肿瘤治疗效果提供理论依据.方法 采用蛋白印迹、流式细胞术、MTT等实验技术,检测TRAIL和低剂量阿霉素分别或者联合处理LNCAP细胞后细胞的生存率变化、相关受体及其配体系统、凋亡相关蛋白(c-FLIP,BCL-2,XIAP)的变化.结果 低剂量阿霉素(0.86 μmol/L)可以增加细胞表面DR4和DR5的表达,同时减少凋亡抑制蛋白c-FLIP的表达,增强TRAIL诱导LNCAP细胞凋亡的敏感性.结论 低剂量阿霉素(0.86 μmol/L)可以增强TRAIL对LNCAP细胞的凋亡诱导效应.     

Effects of sequential treatments with chemotherapeutic drugs followed by TRAIL on prostate cancer in vitro and in vivo

BACKGROUND: Tumor necrosis factor related apoptosis-inducing ligand/Apo2 ligand (TRAIL/Apo-2L) is a novel anticancer agent, capable of inducing apoptosis preferentially in tumor and transformed cells. TRAIL-R1/death receptor (DR)4 and TRAIL-R2/DR5 are members of the tumor necrosis factor (TNF) receptor family, and can be activated by the TRAIL. We examined the clinical potential of chemotherapeutic drugs and TRAIL for the treatment of prostate cancer. METHODS: Prostate and bladder cancer cells were exposed to chemotherapeutic drugs (paclitaxel, vincristine, vinblastine, etoposide, doxorubicin, and camptothecin) and TRAIL. Cell viability was measured by sodium 3'[1-(phenylaminocarbonyl)-3,4-tetrazolium]-bis (4-methoxy-6-nitro) assay; expressions of death receptors and Bcl-2 family members were measured by Western blotting, ELISA and ribonuclease protection assay. PC-3 tumor cells xenografted athymic nude mice were exposed to chemotherapeutic drugs and TRAIL, either alone or in combination, to measure tumor growth and survival of mice. Apoptosis was measured by annexin V-FITC/propidium iodide staining, and terminal deoxynucleotidyltransferase-mediated nick end labeling assay. Caspase-3 activity was measured by the Western blotting and immunohistochemistry. RESULTS: TRAIL induced apoptosis with varying sensitivity. Chemotherapeutic drugs (paclitaxel, vincristine, vinblastine, etoposide, doxorubicin, and camptothecin) significantly augmented TRAIL-induced apoptosis in cancer cells through up-regulation of DR4, DR5, Bax, and Bak, and induction of caspase activation. Mitochondrial pathway enhanced the synergistic interactions between drugs and TRAIL. The sequential treatment of mice with chemotherapeutic drugs followed by TRAIL induced caspase-3 activity, and apoptosis, inhibited angiogenesis, completely eradicated the established tumors, and enhanced survival of mice. CONCLUSIONS: Chemotherapeutic drugs can be used to enhance the therapeutic potential of TRAIL in prostate cancer.     

髂内动脉灌注化疗治疗浸润性膀胱癌

目的探讨浸润性膀胱癌的非手术治疗方法。方法自１９９１年１１月至１９９８年１月应用髂内动脉灌注化疗治疗浸润性膀胱癌２６例,其中１２例行化疗泵植入术和盆腔内动脉再分布术。结果完全缓解率（ＣＲ）２３．１％（６／２６）,部分缓解率（ＰＲ）６９．２％（１８／２６）;副作用较静脉化疗明显减轻。结论此种方法可以作为不能手术切除的浸润性膀胱癌的治疗或作为术前的辅助治疗。