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TwosignaltheoryforTcelactivationprovidedanewapproachfortumorgenetherapy[1-2].Expressionofcostimulatorymoleculegeneintransfect... 相似文献
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重组人内皮抑素腺病毒抑制肝癌裸鼠移植瘤生长 总被引:4,自引:2,他引:4
目的 观察重组人内皮抑素腺病毒(Ad/hEndo)对人肝癌裸鼠移植瘤生长的影响。方法 人脐静脉内皮细胞ECV-304经Ad/hEndo感染后,western印迹检测人内皮抑素的表达。人肝癌BEL-7402细胞移植到裸鼠背脊部后,检测Ad/hEndo对肝癌移植瘤生长的抑制作用。逆转录聚合酶链反应(RT-PCR)检测肿瘤组织中内皮抑素mRNA的表达。分析人内皮抑素在裸鼠体内的表达分布。结果 Western印迹检测到人内皮抑素基因在ECV-304细胞内高效表达。Ad/hEndo明显抑制人肝癌BEL-7402裸鼠移植瘤生长(F=4.061,P<0.05)。Ad/hEndo组血管密度计数为6.88±1.08,DMEM组为13.60±1.71(t=9.216,P<0.01)。瘤内注射Ad/hEndo后3d,RT-PCR在肿瘤组织检测到内皮抑素mRNA的表达,7d后表达不明显。人内皮抑素蛋白主要分布在肿瘤组织。结论 腺病毒介导的人内皮抑素基因在体内、体外获得高效表达,并明显抑制肝癌裸鼠移植瘤的生长与血管生成。 相似文献
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转染人CD80基因的黑色素瘤细胞生长特性及免疫原性探讨 总被引:3,自引:0,他引:3
】 ObjectivesTo investigate the
effects of the CD80 costimulatory molecule expression on the immunogenicity and biological
characteristics of poorly immunogenic B16 melanoma cells,explore the relationship between
the immunogenicity of tumor cells and cell surface molecules. MethodsHuman
CD80 cDNA was transfected into B16 melanoma cells by lipofectin-mediated gene transfer
before the expansion of tumor cells were tested by MTT method in vitro;C57BL/6 mice were
inculated subcutaneously either mock-transfected B16 cells (B16-neo)or CD80-transfected
B16 cells (B16-CD80),then the latency,survival times and tumor mass growth were
investigated.The lymphocytes were examined for both proliferation indices (PI) and
specific cytotoxic activity by MTT method and improved LDH-releasing method,after syngenic
Mixed Lymphocyte tumor cultures (MLTCs). ResultsIn the B16-CD80
cells,in which CD80 expression were detected by SABC method, demonstrated slower growth
rate than B16-neo Clles in vivo (P<0.05),though they didn't in vitro. Compared with
B16-wt and B16-neo cells,B16-CD80 cells more effectively induced the proliferation of
effector lymphocytes and the generation of specific lytic activity against B16-wt cells. ConclusionsThe
CD80 (B7-1) expression in poorly immunogenic tumor cells can increase their immunogenicity
and decrease their tumorigenicity,the effects are associated with the expression of ICAM-1
and MHC molecules on tumor cells surface. 相似文献
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HSV-TK/CD基因联合IL-12基因体内治疗肝癌 总被引:7,自引:0,他引:7
目的观察HSV-TK/CD基因联合IL-12基因体内治疗肝癌的疗效.方法SCID小鼠双侧腋部皮下植入5×105(0.2ml)HepG2细胞,同时腹腔注射PBL 2×107(0.5 ml).荷瘤小鼠分为4组,左侧腋部瘤内分别注射5×105(0.1 ml)丝裂霉素处理过的HepG2-IL-12细胞、5×105(0.1 ml)HepG2-pALBeAFP-CD/TK细胞或两者(各0.05 ml),次日起腹腔注射PBS或GCV 100 mg/kg 5-FC 500 mg/kg,连续10天.Ⅰ组:HepG2-IL-12 GCV,5-FC;Ⅱ组:HepG2-pALBeAFP-CD/TK PBS;Ⅲ组:HepG2-pALBeAFP-CD/TK GCV,5-FC;Ⅳ组:HepG2-pALBeAFP-CD/TK HepG2-IL-12 GCV,5-FC.治疗结束5天后作常规肿瘤组织病理学检查.结果与Ⅱ组比较,Ⅰ、Ⅲ、Ⅳ组治疗侧均有明显的生长抑制,第30天较Ⅱ组分别减小20.5%、54.9%和87.6%,且Ⅰ、Ⅲ、Ⅳ组之间差异明显.Ⅰ、Ⅲ、Ⅳ组右侧肿瘤较Ⅱ组同侧显著缩小,分别达23.2%、24.9%和45.0%.结论pALBeAFP-CD/TK双自杀基因体内治疗肝癌有效.在免疫活性鼠,局部转染IL-12基因可增强双自杀基因的疗效和远处旁观者效应. 相似文献
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人神经营养素-3基因重组腺病毒载体的构建、表达和生物活性检测 总被引:4,自引:0,他引:4
目的构建具有生物活性的人神经营养素-3(NT-3)基因重组腺病毒表达载体.方法从人脑组织mRNA中扩增NT-3基因全长cDNA,定向克隆于穿梭质粒pShuttle中,获得一个带有CMV启动子的质粒.再与腺病毒骨架DNA(Adeno-X viral DNA)体外连接,形成重组腺病毒质粒(pAd-NT-3).用pAd-NT-3转染人胚肾293细胞后包装成有感染能力的重组腺病毒颗粒(Adeno-NT-3). 结果NT-3基因RT-PCR扩增产物约801 bp.Adeno-NT-3 PCR鉴定为正确重组子.RT-PCR、免疫细胞化学、ELISA及Western blot检测显示,Adeno-NT-3能在其转染的293细胞表达和分泌NT-3.这种NT-3能使体外培养的神经干细胞更多地分化为神经元样细胞. 结论应用体外连接法成功构建了人NT-3基因重组腺病毒表达载体,其表达产物具有促进神经干细胞分化为神经元样细胞的活性作用. 相似文献
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目的探讨E1B55KDa缺陷腺病毒dl1520体外对鼻咽癌细胞的杀伤作用。方法dl1520感染不同鼻咽癌株后测定病毒增殖;观察细胞病变效应;MTT法测定细胞生长抑制作用。结果dl1520在鼻咽癌细胞中可复制增殖并有致细胞病变效应,MTT法测定感染复数为0.032、0.16、0.8、4、20、100时CNE2细胞生长抑制率分别为2.78%、7.41%、10.19%、24.07%、45.37%、67.59%。结论dl1520体外可在鼻咽癌细胞中复制并裂解杀伤肿瘤细胞。 相似文献
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