联合运用Survivin和VEGF反义寡核苷酸对裸鼠人肺腺癌A549移植瘤的生长抑制作用  

Survivin antisense oligonucleotide combined with VEGF antisense oligonucleotide inhibits growth of human pulmonary adenocarcinoma A549 xenografts in nude mice

作　　者：梁超[1] 徐玲[1] 隋雪梅[1] 

机构地区：[1]重庆医科大学附属第一医院呼吸内科,重庆400016

出　　处：《第三军医大学学报》2013年第24期2643-2647,共5页Journal of Third Military Medical University

基　　金：国家临床重点专科专项(卫办医政函[2012]949号);重庆市科技攻关计划(CSTC2011AC5202)~~

摘　　要：目的探讨Survivin反义寡核苷酸联合血管内皮生长因子(VEGF)反义寡核苷酸在裸鼠体内对人肺腺癌细胞A549的生长抑制作用。方法将脂质体包裹的ASODN混悬液瘤旁注射移植有A549肿瘤细胞的裸鼠,并相应地把裸鼠分为5组:空白对照组、脂质体对照组、VEGF处理组、Survivin处理组,Survivin-VEGF联合处理组。观察各组肿瘤的生长情况,并分别以Western blot、real-time PCR检测肿瘤组织Survivin及VEGF各自的表达情况,用免疫组化法检测Cyclin B1的蛋白表达和微血管密度计数(CD34染色)。结果 Survivin-VEGF ASODN联合处理组较各组的生长情况明显减慢(P<0.05);Western blot和real-time PCR结果显示Survivin-VEGF联合处理组中Survivin和VEGF蛋白及RNA的表达较各组均受到明显抑制(P<0.05);Survivin-VEGF联合处理组显著降低了肿瘤组织微血管密度计数(MVD)(10.27±1.75),与各单反义寡核苷酸处理组[Survivin处理组(13.27±2.19),VEGF处理组(11.40±1.18)]之间的差异具有统计学意义(P<0.05);Survivin-VEGF联合处理组停滞于G2/M期的细胞(23.17±5.45)也较Survivin处理组(12.54±6.67)和VEGF处理组(18.39±6.87)明显增多(P<0.05)。结论联合使用VEGF反义寡核苷酸和Survivin寡核苷酸对肿瘤的抑制作用优于单用VEGF反义寡核苷酸或Survivin反义寡核苷酸。Objective To investigate the inhibitory effect of survivin antisense oligonucleotide （ASODN） combined with VEGF ASODN on the growth of human pulmonary adenocarcinoma A549 xenografts in nude mice. Methods The nude mice transplanted with A549 tumor cells were divided into 5 groups： control group, liposome control group, VEGF ASODN treatment group, survivin ASODN treatment group, and survivin-VEGF combined ASODN treatment group. Liposome-ASODN mixed suspension was peritumorally injected to the nude mice. The tumor growth in all groups were observed, and the expressions of survivin and VEGF in tumor tissues were detected by Western blotting and real-time PCR. Immunohistochemical method was used to detect the expression of cyclin B1 and microvascular density （MVD） count （CD34 staining）. Results Survivin-VEGF combined ASODN treatment inhibited the tumor growth with more significant effects than other groups （P 〈 0. 05 ）. The result of Western blotting and real-time PCR showed that the protein and mRNA expression of survivin and VEGF in the survivin-VEGF combined ASODN treatment group was obviously lower than other groups （ P 〈 0. 05 ）. Survivin-VEGF combined ASODN treatment group reduced the MVD count in tumor tissues （ 10.27 ± 1.75 ） significantly, with stronger effects than the single ASODN treatment groups （survivin ASODN treatment group 13.27 ± 2. 19 and VEGF ASODN treatment group 11.40 ±1. 18 ） （P 〈0. 05 ）. Survivin-VEGF combined ASODN treatment group group had more cells arrested in in G2/M phase （23.17 ±5.45） than the survivin ASODN treatment group （12.54 ±6.67） and VEGF ASODN group （18.39 ±6.87） （P 〈 0.05）. Conclusion The combination of VEGF the tumor growth of human pulmonary adenocarcinoma A549 xenografts VEGF ASODN or survivin ASODN alone. ASODN in nude and survivin ASODN inhibits mice, with better effects than

关 键 词：SURVIVIN 血管内皮生长因子 反义寡核苷酸 联合 A549 

分 类 号：R394-33[医药卫生—医学遗传学] R73-262[医药卫生—基础医学]