荷瘤小鼠瘤苗治疗疗效与红细胞天然免疫活性相关性及其意义

新近利用抗原处理细胞(APC细胞)处理制备瘤疫效疗较为肯定,认为天然免疫细胞DC细胞瘤疫更有其应用价值.红细胞也是天然免疫细胞.能否用红细胞处理补体调理过的肿瘤细胞制备瘤苗是值得探讨的.有的学者认为红细胞是T细胞活性的调节器,我们的实验有可能为此提供的依据[Arosa FA, Currpharm Des, 2004, 10(27): 191].     

冬虫夏草对肿瘤患者红细胞天然免疫粘附肿瘤细胞的增强作用

[目的]研究药物作用与红细胞天然免疫粘附功能的相关性.[方法]采用红细胞天然免疫粘附酵母菌血凝集和肿瘤红细胞花环试验,测定冬虫夏草的增强作用.[结果]冬虫夏草处理组肿瘤红细胞花环率(24.0±7.5)明显高于未处理组(10.8±3.2,P＜0.01).[结论]试管内冬虫夏草能改善肿瘤患者的红细胞天然免疫活性.     

抗膀胱癌免疫毒素的构建及表达

为了对膀胱癌进行早期的检测和治疗,运用免疫毒素寻找并有效杀死癌细胞不失为一个好方法.但是,早期的免疫毒素是通过化学偶联而成的,有以下缺点:1.易释放从而伤害了正常细胞.2 免疫毒素的免疫原性较强.且渗透性较差,导致毒素的活性不够强.为了克服上述缺点,我们利用DNA重组技术,将异源的SeFv基因与削减的毒素基因融合并克隆,在大肠杆菌中翻译出单链融合蛋白.这就是所谓的第三代免疫毒素,又称基因工程免疫毒素.     

免疫毒素与顺铂协同杀伤肿瘤细胞机制的探讨

背景与目的:免疫毒素HELβ-PE38KDEL(HeregulinEGFlikedomainβ1-PsedomonasAeurginosa38KDEL,本文简称ITH)已经被证实是一种具有特异性杀伤erbB2、3、4阳性乳腺癌细胞的新的生物学制剂,然而它与化疗药物的联合作用效果目前尚未有报道。本文探讨ITH与化疗药物顺铂(DDP)的协同抗肿瘤作用。方法:采用AnnexinV结合实验和Westernblot检测乳腺癌细胞MDA-MB-453、2LMP和胃癌细胞N87分别在ITH和DDP单独及联合用药前后的细胞凋亡变化。结果:在高表达erbB-2、3、4的MDA-MB-453和N87中,联合用药组和单药组相比,凋亡细胞成倍增加(P<0.01);而在低表达erbB-2、3、4的2LMP,联合用药组和单药组相比,凋亡细胞无明显增加(P>0.05)。MDA-MB-453和N87细胞在联合用药组中PARP、caspase-3降解增加,bcl-2、mp53过度表达受抑制;而2LMP细胞中PARP、caspase-3降解无增加,bcl-2、mp53过度表达未受抑制。结论:ITH和DDP联合后可促进高表达erbB-2、3、4的乳腺癌细胞凋亡,这可能是两者协同作用的机制之一。     

放射性碘标抗膀胱癌免疫毒素生物分布与代谢研究

目的: 对偶联苦瓜毒素(MT) 后131I-BDI-1在荷瘤裸鼠体内分布与代谢性能变化进行研究,探讨其应用于膀胱癌导向治疗的可能性.方法: MT与BDI-1通过异型双功能交联剂SPDP进行偶联;BDI-1与BDI-1-MT采用ChT法进行131I标记;两组荷人膀胱癌裸鼠分别经尾静脉注射131I-BDI-1-MT与131I-BDI-1,经48 h, 72 h及120 h后进行体内放射性分布测定与γ显像.计算肿瘤与各个脏器的摄取百分数(%ID/g)及与正常组织放射性之比(T/NT).结果: 与131I-BDI-1 相比,131I-BDI-1-MT在肿瘤中的摄取率未见明显降低,但在多数正常组织中的摄取率显著低于131I-BDI-1,其T/NT值明显高于131I-BDI-1;131I-BDI-1-MTγ影像中肿瘤与正常组织的对比度优于131I-BDI-1;131I-BDI-1-MT与131I-BDI-1在肿瘤中清除速率差异不大,而131I-BDI-1-MT在正常组织中(尤其在血液中)的清除速率高于131I-BDI-1.结论: 131I-BDI-1-MT有抗肿瘤活性,可有效地靶向膀胱癌细胞.     

免疫毒素抗膀胱癌的实验与初步临床应用

目的:探讨抗人膀胱癌单克隆抗体绿脓杆菌外毒素(BDI-1-PEA)对膀胱癌细胞的特异杀伤作用。方法:应用四甲基偶氮唑盐(MTT)法检测10-10～10^-7mol/L浓度下的BDI-1-PEA、抗人膀胱癌单克隆抗体(BDI-1)、BDI-1和绿脓杆菌外毒素(PEA)的混合物(PEA+BDI-1)对体外癌细胞的杀伤能力。接种于裸鼠背部皮下的癌细胞长至直径0.3cm大小实体瘤时,随机分三组,每组10只裸鼠,于尾静脉隔日分别注入100倍半致死剂量浓度(1×10^-7mol/L)的BDI-1-PEA100μl及相同剂量的BDI-1、正常小鼠IgG共6次,观察不同的药物对癌的抑制作用。24例膀胱移行细胞癌术后患者应用100倍半细胞致死剂量浓度(1×10^-7mol/L)50mlBDI-1-PEA进行了膀胱灌注治疗,每周2次,共8次。每3个月复查膀胱镜或B超。结果:BDI-1-PEA抗膀胱癌细胞系E-J的作用明显优于单抗及毒素与单抗的混合物(P<0.01),与对非靶细胞肾肿瘤细胞系786-0的细胞毒性作用相比较差异非常显著。24例膀胱移行细胞癌术后患者,用药后随访7～22个月,平均15个月,2例患者复发。结论:BDI-1-PEA对膀胱癌细胞具有特异杀伤作用,对预防膀胱癌术后复发有较好疗效。     

插入外源性白细胞介素2基因的肿瘤细胞的抗肿瘤抗转移作用

中国免疫学会肿瘤免疫与生物治疗专业委员于1991年12月经中国免疫学会一届二次理事会批准,1992年5月在河南汤阴召开的第三次筹备会上正式成立.专业委员会由张友会、崔正言、钱振超、郑升、高进、丁仁瑞、刘华、刘新垣、孔宪寿、姜迁良、李殿俊、田志刚、王大庆、曹雪涛、陈家畅、刘旭、樊代明组成,并推选张友会为主任委员,崔正言、钱振超、郑升为副主任委员,秘书由张叔人担任.会议回顾了1989年、1991年分别在大连、济南召开的第一、二届肿瘤生物治疗全国学术会议以来的工作,并决定今后继续每两年举办一次全国学术会议.会议还讨论了创办自己的学术刊物等事项.自汤阴会议以来,专业委员会的工作有了发展,1993年在哈尔滨召开了第三届全国学术会议,并作出积极筹办《中国肿瘤生物治疗杂志》的决定,杂志挂靠在第二军医大学.     

榄香烯对肿瘤细胞生物学特性的影响及其瘤苗免疫效应机制的研究

为研究我们创制的榄香烯(elemene,E)复合瘤苗主动免疫抗瘤效应的分子机制及其与热休克蛋白(heat shock protein,HSP)的可能关系,本文通过光镜、电镜和流式细胞术(FCM)以热休克为比照,研究了抗癌中药有效组分榄香烯对小鼠H22腹水型肝癌(H-2~-)和L615白血病(H-2~k)细胞某些生物学特性(生存率、形态结构、坏死、凋亡、细胞周期等)的影响;在此基础上着重观察比较了:1.榄香烯或/和热休克H22和L615瘤苗的主动免疫保护效应.2.榄香烯复合瘤苗诸因素(榄香烯、MMC、戊二醛)与热休克对两种肿瘤细胞膜HSP70和HSP90表达的影响.3.分离纯化榄香烯、MMC诱导的H22肿瘤细胞的HSP70-肽复合物体内验证其主动免疫预防效应.     

不同时期应用细胞瘤苗对红白血病荷瘤小鼠抗瘤作用的研究

目的：探讨不同时间应用细菌瘤苗对红细胞病荷瘤小鼠的抗瘤作用。方法：在不同时间内应用瘤苗免疫治疗红白血病荷瘤小鼠，观察小鼠生存期，皮下肿瘤大小及肿瘤，注射部位病理变化。结果：3天，7天瘤苗治疗的小鼠生存期延长，皮下肿瘤生长缓慢，病理可见瘤组织坏死及大量以单个核细胞为主的炎细胞浸润，与PBS组及10天瘤苗组相比，有显著性差异。结论：3天，7天瘤苗比10天瘤苗组抗肿瘤作用强。     

白血病细胞来源的树突状细胞在治疗白血病中的应用

目的:探讨免疫毒素BDI-1-PEA特异杀伤膀胱癌细胞的体外及体内研究.方法:应用MTT法检测不同浓度的BDI-1-PEA,BDI-1和PEA的混合物(BDI-1 PEA),PEA对体外癌细胞的杀伤能力.接种于裸鼠背部皮下的癌细胞长至0.3cm大小实体瘤时,于尾静脉隔日分别注入BDI-1-PEA、BDI-I、正常小鼠IgG共6次,观察不同的药物对癌的抑制作用.结果:免疫毒素BDI-1-PEA在体外抗膀胱癌细胞系E-J的作用明显优于PEA及BDI-1与PEA的混合物,在荷瘤裸鼠体内明显优于BDI-1及IgG,与对非靶细胞的细胞毒性作用相比较差异非常显著.结论:免疫毒素BDI-1-PEA是一种很有潜力的抗癌药物,为进一步临床研究奠定基础.     

ppGalNAc T1 as a Potential Novel Marker for Human Bladder Cancer

Objectives: To investigate the effect of glycopeptide-preferring polypeptide GalNAc transferase 1 (ppGalNAcT1 ) targeted RNA interference (RNAi) on the growth and migration of human bladder carcinoma EJ cells invitro and in vivo. Methods: DNA microarray assays were performed to determine ppGalNAc Ts(ppGalNAc T1-9)expression in human bladder cancer and normal bladder tissues. We transfected the EJ bladder cancer cell linewith well-designed ppGalNAc T1 siRNA. Boyden chamber and Wound healing assays were used to investigatechanges of shppGalNAc T1-EJ cell migration. Proliferation of shppGalNAc T1-EJ cells in vitro was assessedusing [3H]-thymidine incorporation assay and soft agar colony formation assays. Subcutaneous bladder tumorsin BALB/c nude mice were induced by inoculation of shppGalNAc T1-EJ cells and after inoculation diametersof tumors were measured every 5 days to determine gross tumor volumes. Results: ppGalNAc T1 mRNA inbladder cancer tissues was 11.2-fold higher than in normal bladder tissues. When ppGalNAc T1 expressionin EJ cells was knocked down through transfection by pSUPER-shppGalNAc T1 vector, markedly reducedincorporation of [3H]-thymidine into DNA of EJ cells was observed at all time points compared with the emptyvector transfected control cells. However, ppGalNAc T1 knockdown did not significantly inhibited cell migration(only 12.3%). Silenced ppGalNAc T1 expression significantly inhibited subcutaneous tumor growth comparedwith the control groups injected with empty vector transfected control cells. At the end of observation course (40days), the inhibitory rate of cancerous growth for ppGalNAc T1 knockdown was 52.5%. Conclusion: ppGalNAcT1 might be a potential novel marker for human bladder cancer. Although ppGalNAc T1 knockdown caused noremarkable change in cell migration, silenced expression significantly inhibited proliferation and tumor growthof the bladder cancer EJ cell line.     

人膀胱移行细胞癌内的16／18型人乳头状瘤病毒感染

目的 探讨人膀胱移行细胞癌与高危型人乳头瘤病毒感染的关系。方法 采用聚合酶链式反应（ＰＣＲ法）检测１１２例膀胱移行细胞组织（包括７５例石蜡包埋组织和３７例手术切除组织）和７例正常膀胱粘膜组织的ＨＰＶ－１６／１８感染率及ＨＰＶ－１６／１８感染与膀胱移行细胞癌病理分极及临床分期的关系。同时检测了２４例膀胱癌病人尿液沉淀中ＨＰＶ阳性率。结果 膀胱移行细胞癌的ＨＰＶ－１６／１８的总感染率为６２．５０％（７     

光动力作用对膀胱癌细胞生长影响的体外实验研究

目的观察光动力治疗(PDT)对体外培养的人膀胱癌细胞的杀伤效应以及形态改变,同时探讨光敏剂(PhtofrinⅡ)浓度与杀伤效应的关系。方法将膀胱癌细胞(T-24)分组进行培养,扩增至一定数目后,加入光敏剂,然后用激光照射,绘制细胞生长曲线,观察PDT对肿瘤细胞生长的抑制情况,同时进行细胞形态学观察。结果癌细胞生长随光敏剂用药剂量不同而受到不同程度抑制;显微镜下见治疗后癌细胞结构受到破坏。结论体外光动力治疗可明显抑制膀胱癌细胞的增长,其效应与光敏剂浓度呈正相关。     

Concentration- Dependent Effects of Curcumin on 5-Fluorouracil Efficacy in Bladder Cancer Cells

Purpose: Curcumin (Cur), a herbal ingredient with anticancer properties, has been shown to inhibit growth of malignant cells in vivo and in vitro. However, studies on combination therapy of Cur with chemotherapeutic drugs have been limited. Here, effects of Cur on the cytotoxicity of 5-Fluorouracil (FU) were investigated with epithelial bladder cancer cells (EJ138) in vitro. Methods: EJ138 cells were treated with 5 and 15 μM of Cur and/ or 100 μM of FU. Cell viability was measured by sulforhodamine B colorimetric assay. The glucose concentration as an index of cell metabolism was evaluated by an enzymatic method. Total oxidant and antioxidant capacities were estimated by the ferrous oxidation-xylenol (FOX1) method and ferric reducing antioxidant power assay (FRAP), respectively. Results: Combination of 5 μM Cur with FU significantly reduced its cytotoxicity in EJ138 cells, while 15 μM Cur caused an opposite increase. Significant increase in glucose concentration at 24 h and decrease in the FRAP value at 48 h incubation was observed in cells treated with FU in combination with Cur. There were no significant changes in total oxidant capacity with the combination therapy. Conclusion: Our findings suggest a crucial role of Cur concentration in regulating chemotherapeutic agent-induced cytotoxicity. Further investigations are needed to understand the precise mechanisms of action of Cur and determine appropriate doses with combination therapy for clinical application against human cancers.     

147例表浅性膀胱癌预后因素分析

目的 :探讨表浅性膀胱癌各种因素与患者预后的关系。方法 :对 147例表浅性膀胱癌进行回顾性分析。结果 :147例中 ,72例术后复发 ( 4 9% ) ,术后 5年复发率为 35.4 %。初诊时为多发者、直径大于 3cm、分级与分期高的肿瘤术后复发率分别高于单发者、直径小于 3cm者、分级、分期低的肿瘤。术后 6个月内肿瘤复发者经治疗后肿瘤再次复发的机会高 ,术后膀胱内灌药可以预防肿瘤复发。结论 :肿瘤分级与分期高、多发肿瘤、直径大于 3cm者及术后膀胱内未灌药者复发率高     

Expression of miR-143 Reduces Growth and Migration of Human Bladder Carcinoma Cells by Targeting Cyclooxygenase-2

Systemic chemotherapy is the only current modality that provides the potential for long-term survivalin bladder carcinoma patients with metastatic disease. Overexpression of cyclooxygenase-2 COX-2 inducesexpression of immune- and cell proliferation-related genes and is associated with the grade, prognosis andrecurrence of transitional cell carcinoma of the bladder. There is abundant evidence that aberrant expressionof microRNAs (miRNAs) is implicated in numerous disease states and miRNAs have the potential to be used forcancer therapeutics. Here, we found expression of miR-143 to be low in a series of human bladder carcinomas ascompared to background tissue. In addition, restoration of miR-143 by cell transfection in T24 cancer cells led todecreased COX-2 expression, reduced proliferation and mobility. Our findings will help to further understand thefunctions of miRNAs in cancer cells and point to a specific potential of miR-143 may be employed as a therapeuticagent for bladder carcinoma. The results provide insights into the development of novel tumor markers or newtherapeutic strategies.     

Preparation of Immunotoxin Herceptin-Botulinum and Killing Effects on Two Breast Cancer Cell Lines

Background: Worldwide, breast cancer is the most common cancer diagnosed among women and a leadingcause of cancer deaths. The age of onset in Iran has become reduced by a decade for unknown reasons. Herceptin,a humanized monoclonal antibody, is a target therapy for breast cancer cells with over expression of HER2-neu receptors, but it is an expensive drug with only 20% beneficial rate of survival. This study introduces anovel approach to enhance the efficacy of this drug through immunoconjugation of the antibody to botulinumtoxin. Decreasing the cost and adverse effects of the antibody were secondary goals of this study. Materials andMethods: Botulinum toxin was conjugated with Herceptin using heterobifunctional cross linkers, succinimidylacetylthiopropionate (SATP) and sulfo-succinimidyl-4-(N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC)according to the supplier’s guidelines and tested on two breast cancer cell lines: SK-BR-3 and BT-474. Toxinand Herceptin were also used separately as controls. The cytotoxicity assay was also performed using the newbioconjugate on cultured cells with Alamar blue and a fluorescence plate reader. Results: Herceptin-Toxinbioconjugation significantly improved Herceptin efficacy on both breast cancer cell lines when compared tothe control group. Conclusions: Toxin-Herceptin bioconjugation can be a potential cand     

NQO1基因多态性与膀胱癌易感性的关系

[目的]探讨浙江地区依赖还原型辅酶Ⅰ/Ⅱ醌氧化还原酶1(NQO1)基因多态性与膀胱癌易感性的关系.[方法]采用病例对照研究方法,应用相对的两对引物—聚合酶链反应技术(PCR-CTPP),对99例膀胱癌患者和100例非肿瘤患者的NQO1 C609T基因型进行检测,并分析其与膀胱癌易感性的关系.[结果]NQO1 C609T基因型分布频率在膀胱癌组和对照组之间的差异具有统计学意义(P＜0.05),携带609TT基因型的个体罹患膀胱癌的风险是携带609CC基因型个体的2.448倍(95％CI为1.125～5.326).NQO1 C609T基因型分布频率在膀胱癌不同病理分级和临床分期之间的差异均无统计学意义(P＞0.05).[结论]NQO1 C609T基因多态性可能与膀胱癌的易感性相关,携带NQO1 609TT基因型的人群易患膀胱癌.