利用siRNA沉默VEGF对膀胱癌生长的影响  

siRNA inhibits vegf expression in bladder cancer

作　　者：李然伟[1] 王秀岩[1] 杨泽成[2] 郭万松[3] 刘辉[4] 赵燕颖[5] 

机构地区：[1]吉林大学第二医院泌尿外科,吉林长春130041 [2]吉林大学中日联谊医院普外科,吉林长春130033 [3]长春市中心医院泌尿外科 [4]吉林省人民医院,吉林长春130021 [5]吉林大学基础医学院生理学教研室,吉林长春130021

出　　处：《中国实验诊断学》2008年第2期188-192,共5页Chinese Journal of Laboratory Diagnosis

基　　金：吉林省科委社会发展项目(200505118)

摘　　要：目的研究VEGF特异性RNA干扰对膀胱癌细胞体内外增殖的影响。方法针对VEGF构建siRNA真核表达载体PsilencerTM-VEGF-siRNA,并转染至膀胱癌细胞T24。RT-PCR观察VEGF的基因沉默效果;MTT比色分析检测细胞体外增殖能力;流式细胞计数检测细胞的凋亡。同时,通过裸鼠移植瘤模型观察VEGF基因沉默效果及对膀胱癌体内治疗的效果。结果转染siVEGF后膀胱癌细胞的VEG基因的表达显著被抑制(最高抑制率达79%)。与对照组比较,转染siVEGF组T24细胞的增殖活性明显低于转染阴性对照质粒组和空白对照组(P<0.01);流式细胞计数结果显示细胞发生凋亡。裸鼠体内致瘤实验显示,转染siVEGF组T24细胞在裸鼠体内增殖显著降低,肿瘤生长减慢。结论siVEGF能够有效的抑制膀胱癌中VEGF的表达,从而有效抑制膀胱癌细胞的生长。Objective To investigate the inhibition effect of the RNA interfere targeting VEGF on the growth of bladder cancer cell in vitro and in vivo.Methods The siRNA eukaryotic expression vector targeting VEGF gene,named Psilencer^TM-VEGF-siRNA was constructed and transfected into T24 cell. The inhibition effects on VEGF gene were determined by reverse transcription PCR analysis. The invitro cellular growth activities were assayed by MTT colorimetry. The cell apoptosis was studied boflow cytometry. TO observed the therapeutic effects of siRNA VEGF through xenograft tumor nude mice model. VEGF expression was analyze with RT-PCR and Western blot. Results After the siVEGF transfected into the T24 cells, the expressio of VEGF gene was inhibited significantly （by 79% ）.The cellular growth activities in the T24 cells transfected with siVEGF decreased obviously,significantly lower than those in the regative control group and in the nontransfected group（ P 〈 0.01）.Significantly apoptosis in the T24 cells transfected with siVEGF was induced. The growth speed and formation rate of xenograft tumor in siRNA VEGF transfected mice slowed down significantly, and VEGF expression were down regulated too. However, there was no similar inhibitive effect in the control groups. Conclusion siRNA targeting VEGF gene inhibit VEGF expression and inhibits the growth of bladder cancer.

关 键 词：SIRNA 膀胱癌 VEGF 凋亡 

分 类 号：R737.14[医药卫生—肿瘤]