靶向EBV-LMP1脱氧核酶对鼻咽癌裸鼠移植瘤生长的影响
Effect of DNAzyme on expression of LMP1 gene on human nasopharyngeal carcinoma xenograft in nude mice
柯霞 1杨玉成 1洪苏玲1
作者信息
- 1. 重庆医科大学附属第一医院耳鼻咽喉科,重庆,400016
- 折叠
摘要
目的:观察EBV-LMPl基因的靶向脱氧核酶(DZ)对鼻咽癌裸鼠移植瘤牛长的影响.方法:设计并合成针对EBV-LMP1基因的脱氧核酶DZ509,突变型脱氧核酶mutDZ509及其反义寡核苷酸ASODN509.裸鼠皮下注射鼻咽癌细胞(C666-1),建立皮下移植瘤模型,肿瘤形成后分为瘤内注射DZS09 60 μmol/L,mutDZ509 60 μmol/L,ASODN509 106 μmol/L,PBS 50 μL;记录肿瘤生长曲线,采用半定量RT-PCR法检测LMPl mRNA表达,免疫组化方法,Western Blot检测LMPI蛋白表达.结果:成功构建裸鼠皮下鼻咽癌移植瘤模型.DZS09组,mutDZ509组,ASODN509组与对照(PBS)组相比较,肿瘤生长速度依次减慢,3组抑瘤率分别为85.53%,73.03%,52.77%.LMPl mRNA及蛋白表达依次增加,差异具有统计学意义(P<0.05).结论:瘤体内注射DZ能有效抑制LMPl表达并抑制鼻咽癌移植瘤牛长,针对EBV-LMPl的DZ可能成为鼻咽癌治疗的基因药物.
Abstract
AIM: To observe the inhibitory effect of DNAzyme targeting on Epstain-Barr virus encoded latent membrane protein 1 (EBV-LMP1) on the growth of nasopharyngeal carcinoma ( NPC) xenograft in nude mice. METHODS: We designed and synthesized DNAzyme ( DZ509 ) , mutant DNAzyme ( mutDZ509 ) and antisense oligonucleotides ( ASODN509) targeting on EBV-LMP1; nasopharyngeal carcinoma cell (C666-1 cell) was injected into 28 nude mice subcutaneously to establish cancer model and then divided into 4 groups including DZ509, mutDZ509, ASODN509 and PBS. We described growth curve of tumor, tumors were cutting after 8 d, detected LMP1 mRNA by reverse transcription-PCR (RT-PCR), and LMP1 by immunohistochemistry and Western Blot. RESULTS: NPC cancer models with nude mice were established; Among DZ509,mutDZ509,ASODN509 and PBS, the growth velocity of tumor stepped down, the rate is 85. 53% ,73. 03% , 52.77%,and the expression of LMP1 mRNA and LMP1 protein increased in turn. CONCLUSION; Injecting DNAzyme into tumors in vivo can inhibit the express of LMP1 and growth of nasopharyngeal carcinoma xenograft effectively. DNAzyme targeting on LMP1 may be a new promising gene therapy for NPC.
关键词
脱氧核酶/潜伏膜蛋白1/裸鼠/鼻咽癌/EB病毒Key words
DNAzyme/LMP1 gene/nude mice/nasopharyngeal carcinoma/Epstein-Barr virus引用本文复制引用
出版年
2009
NETL
NSTL
维普
万方数据