Survivin反义寡核苷酸对小鼠肺癌的生长抑制作用  

Survivin Antisense Oligonucletides in Murine Lung Cancer

作　　者：巴一[1] Reisfeld Ralph A. 

机构地区：[1]南京医科大学附属第一临床医学院肿瘤科 [2]Dept.of Immunology The Scripps Research Institute,LaJolla,CA,USA92037

出　　处：《同济大学学报（医学版）》2005年第2期1-4,共4页Journal of Tongji University(Medical Science)

基　　金：国家自然科学基金资助项目 (3 0 40 0 5 3 8)

摘　　要：目的　Survivin是一种凋亡抑制蛋白(inhibitorofapoptosisproteins ,IAP)家族成员,在几乎所有肿瘤组织中特异性表达,而在正常成年终末分化组织中低表达甚至不表达。选择五组小鼠survivin反义寡核苷酸,并从中进步筛选效果显著的反义寡核苷酸,在小鼠体内进一步验证其抑制肿瘤生长的能力。方法　设计合成特异性靶向生存素的反义寡核苷酸五组。利用脂质体转染技术将其转入肺癌细胞D12 1,Westernblot检测survivin的表达抑制效应,从中筛选效果显著的反义寡核苷酸,确定最佳浓度,将此浓度下转染的D12 1细胞经小鼠尾静脉注入,观察生存曲线。结果　各组反义寡核苷酸在4 0 0nmol·L-1转染细胞,Westernblot结果显示NO .5组survivin表达程度减弱,选取NO .5反义寡核苷酸,在不同浓度下观测转染2 4h后,细胞凋亡情况及survivin表达状况。2 0 0nmol·L-1时,sur vivin表达出现减弱,但细胞生长尚未受到影响;4 0 0nmol·L-1时细胞开始出现凋亡,指数明显高于对照组(P <0 .0 5 ) ,以6 0 0nmol·L-1转染组最为明显。用NO .5反义寡核苷酸在2 0 0nmol·L-1浓度下转染D 12 1细胞,将此细胞致瘤小鼠,小鼠的生存期明显延长。结论　因为survivin基因在多数恶性肿瘤组织中丰富表达,选择合适的sur vivin反义寡核苷酸。Objective Survivin,a recently described inhib it or of apoptosis,is expressed in cancer cells and not in normal adult tissues,an d is recognized as a potential target in anticancer therapy.Five antisense oligo nucleotides (ASON) were designed and selected.This study was aimed to investigat e the effect of antisense oligonucleotide targeting survivin on murine lung canc er.Methods Five (ASON) targeting survivin were designed and transfected into D121 cells.After transfection for 24 h,cultured cells were harv ested and survivin protein expression was detected by Western blot.Selected ASOD N was used for the transfection and mice challenge. Results The experiment results demonstrated that antisens e oligonucleotide No.5 could reduce survivin protein level obviously.Expression of survivin was decreased at 200 nmol·L -1 in which cell morphological change was not found.From a concentration of 400 nmol· L -1,survivin was decreased as compared with that in the control and abnormal morphological change of cells was observed at the same time.200 nmol ·L -1 No.5 antisense oligonucleotide was used for tumor targ eting in vivo,life survival was significantly improved in mice. Conclusion The selected expression of survivin in tumor c ells may provide a high degree specificity for cancer therapy using survivin ant isense oligonucleotide.

关 键 词：SURVIVIN 反义寡核苷酸 凋亡 小鼠 

分 类 号：R734.2[医药卫生—肿瘤]