靶向VEGF基因的siRNA体外抑制兔VX2细胞生长的研究  

The study of VEGF-targeted siRNA in vitro inhibiting the growth of rabbit VX2 cell

作　　者：邹煜[1,2] 过川根[2] 羊正纲[3] 张敏鸣[4] 

机构地区：[1]浙江大学医学院附属妇产科医院放射科,浙江杭州310006 [2]浙江大学医学院附属第一医院放射科,浙江杭州310003 [3]浙江大学医学院附属第一医院传染病研究所,浙江杭州310003 [4]浙江大学医学院附属第二医院放射科,浙江杭州310009

出　　处：《现代肿瘤医学》2015年第19期2720-2723,共4页Journal of Modern Oncology

基　　金：浙江省自然科学基金项目(编号:Y2110172);浙江省教育厅科研基金项目(编号:Y201225858)

摘　　要：目的:探讨靶向VEGF基因的siRNA对兔VX2肿瘤细胞生长的影响。方法:采用化学合成法得到靶向兔VX2细胞VEGF基因的siRNA分子,应用脂质体将其转染体外培养的VX2细胞。转染48h后,RT-PCR法检测VEGF mRNA水平,ELISA法检测细胞分泌的VEGF蛋白,MTT法检测细胞生长活力。结果:VEGFsi1和VEGF-si3单独转染VX2细胞,细胞的生长抑制率分别为(38.5±7.3)%和(30.0±5.8)%,均显著高于scr-siRNA转染的对照组(P<0.01);细胞VEGF mRNA表达水平分别为(0.37±0.06)和(0.45±0.07),均显著低于对照组(P<0.01);细胞分泌VEGF蛋白的抑制率分别为(58.1±7.3)%和(51.9±7.9)%,均显著高于对照组(P<0.01)。结论:VEGF siRNA分子在体外能特异地抑制兔VX2细胞的VEGF基因mRNA转录,VEGF蛋白分泌以及细胞增殖。Objective：To investigate the effect of small interfering RNA（siRNA）targeting vascular endothelial growth factor（VEGF）on the growth of rabbit squamous epithelial cancer cell line VX2 cells. Methods：Three groups of siRNA targeting VEGF gene in VX2 cells（VEGF - si1,VEGF - si2,VEGF - si3）were designed and obtained by chemical synthesis. Then these siRNAs were transfected into cultured VX2 cells in vitro by Lipofectamine. At 48 hours after VEGF - siRNA transfection,the levels of VEGF mRNA expression were analyzed by reverse transcription poly-merase chain reaction（RT - PCR）,the secretion of VEGF protein in the supematant was examined by ELISA,and the viability of VX2 cells was detected by MTT. Results：For VEGF - si1 or VEGF - si3 transfected VX2 cells,The growth of cells was inhibited by the rate of（38. 5 ± 7. 3）% or（30. 0 ± 5. 8）% ,significantly higher than that of scramble siRNA transfected VX2 cells（control group）,（P 〈 0. 01）;The level of VEGF mRNA was（0. 37 ± 0. 06）or （0. 45 ± 0. 07）,significantly lower than that in control group,（P 〈 0. 01）;The secretion of VEGF protein was also decreased by the rate of（58. 1 ± 7. 3）% or（51. 9 ± 7. 9）% ,notably higher than that of control group,（P 〈 0. 01）. Conclusion：VEGF - targeted siRNA can in vitro specifically inhibit VEGF mRNA transcription,VEGF protein secre-tion and the proliferation of VX2 cell line.

关 键 词：小干扰RNA 血管内皮细胞生长因子 VX2细胞 转染 

分 类 号：R73[医药卫生—肿瘤]