靶向存活素的RNAi联合X射线照射对肺腺癌移植肿瘤的抑制作用  

Inhibitory effects of RNAi targeting Survivin combined X-rays irradiation on grafting tumour of lung adenocarcinoma

作　　者：李长锋[1] 张斌[1] 张金萍[1] 王志成[2] 杨海山[1] 

机构地区：[1]吉林大学中日联谊医院,吉林长春130033 [2]吉林大学公共卫生学院卫生部放射生物学重点实验室

出　　处：《中国实验诊断学》2009年第4期430-432,共3页Chinese Journal of Laboratory Diagnosis

基　　金：吉林省科技厅资助项目(200705274)

摘　　要：目的利用RNAi、RT-PCR和Western blot技术,探讨靶向存活素的基因沉默联合X射线照射对肺腺癌移植瘤的抑制作用及相关机制。方法根据Genbank中存活素(Survivin)的cDNA序列设计RNAi序列,构建shRNA真核表达载体pGenesil-2-Survivin,经过酶切,测序鉴定后,将经脂质体包裹的pGenesil-2-Survivin注射到裸鼠移植肺腺癌瘤体内,并联合5 Gy X射线照射,利用RT-PCR和Western blot技术检测移植肿瘤内Survivin mRNA和蛋白表达情况,观察肿瘤生长变化。实验分为正常组、pGenesil-2-Survivin组、5 Gy照射组和pGenesil2-Survivin+5 Gy组。结果RT-PCR和Western blot结果显示,5 Gy X射线没有明显改变Survivin mRNA和蛋白的表达,而pGenesil-2-Survivin和pGenesil-2-Sur-vivin+5 Gy照射则明显抑制了Survivin的表达,与正常组比较具有统计学意义(P<0.001);5 Gy照射后,肿瘤的生长被抑制,而pGenesil-2-Survivin质粒的抑制作用更强,5 Gy照射能增强pGenesil-2-Survivin质粒的抑制作用。结论靶向存活素的RNAi能够明显抑制裸鼠移植肿瘤的生长,联合5 Gy X射线照射能够增强抑制作用,这可能与Survivin基因mR-NA与蛋白被抑制相关。Objective To approach the inhibitory effects of RNAi targeting Survivin combined X-rays irradiation on grafting tumour of lung adenocareinoma and realative mechanism with RNAi, RT-PCR mM Westren blot. Methods To design one pair RNA interference sequence targeting Survivin gene according to cDNA sequence showed in GeneBank, to construct the shRNA eukaryotic expressing vector pGenesil-2-Survivin. Normal group, pGenesil-2-Survivin group, 5 Gy irradiation group and pGenesil2-Survivin ＋ 5 Gy irradiation group were divided.Results RT-PCR and Western blot results showed that 5 Gy X-rays could not change Survivin mRNA and protein expressions obviously,but pGencsil-2-Survivin plasmid or pGenesil-2-Survivin plasmid adding 5 Gy X-ray irradiation could inhibit Survivin mRNA and protein expressions obviously,compared to normal group, it had statistical significance （ P 〈 0.001 ）;after 5 Gy X-rays irradiation, tumour growth was inhibited,but pGenesil-2-Survivin plasmid inhibitory effects was more obvious,and 5 Gy X- rays irradiation could enhanced the effect. Conclusion RNAi targeting Survivin could inhibit obviously the growth of nude mouse grafting tumour of lung adenocarcinoma, and combination with 5 Gy X-rays irradiation could enhance the inhibitory effects, these related to Survivin mRNA＂ and protein expression.

关 键 词：RNA干扰 生存素 X射线照射 抑制作用 

分 类 号：R734.2[医药卫生—肿瘤]