VEGF反义寡核苷酸对胆囊癌细胞VEGF，Flt-1及KDR mRNA表达和VEGF蛋白分泌的影响  

Effects of vascular endothelial growth factor antisense oligodeoxynucleotide on the mRNA expression of vascular endothelial growth factor,fms-Iike tyrosine kinase-1 and kinase insert domaincontaining receptor and vascular endothelial growth factor protein

作　　者：李海军[1] 庞作良[1] 毛拉艾沙.买买提 

机构地区：[1]新疆医科大学附属肿瘤医院肝胆胰外科,新疆省乌鲁木齐市830011

出　　处：《世界华人消化杂志》2007年第11期1225-1231,共7页World Chinese Journal of Digestology

摘　　要：目的:体外研究Oligofectamine介导的VEGF反义寡核苷酸(antisense oligodeoxynucleotide,ASODN)转染对人胆囊癌GBC-SD细胞VEGF,Flt-1及KDR mRNA表达和VEGF蛋白分泌的影响.方法:运用Oligofectamine介导的VEGF反义寡核苷酸(ASODN)和错义寡核苷酸(Scrambled Oligodeoxynucleotide,SODN)转染人胆囊癌细胞GBC-SD,半定量RT-PCR检测转染后各组细胞不同时间的VEGF,Flt-1及KDR mRNA表达变化,ELISA测定转染后各组细胞培养上清液VEGF蛋白浓度.结果:半定量RT-PCR发现ASODN组及ASODN+Oligofectamine组24,48,72,96 h VEGF(ASODN组VEGF165:0.686±0.033,0.569±0.049,0.489±0.036,0.716±0.017;ASODN组VEGF121:0.462±0.046,0.338±0.034,0.219±0.022,0.471±0.038;ASODN+Oligofectamine组VEGF165:0.601±0.021,0.465±0.042,0.416±0.023,0.662±0.035:ASODN+Oligofectamine组VEGF121:0.408±0.014,0.286±0.019,0.157±0.021,0.418±0.037)、Flt-1(ASODN组:0.694±0.019,0.562±0.045,0.435±0.042,0.724±0.026;ASODN+Oligofectamine组:0.609±0.018,0.442±0.049,0.314±0.015,0.614±0.029)及KDR(ASODN组:0.667±0.063,0.490±0.033,0.301±0.029,0.665±0.068;ASODN+Oligofectamine组:0.523±0.048,0.432±0.027,0.218±0.036,0.524±0.037) mRNA的表达显著低于Control组(P<0.05),且ASODN+Oligofectamine的抑制作用比ASODN强(P>0.05).ELISA测定结果显示ASODN组(281.26±18.62,526.44±34.95,791.13±20.99)及ASODN+Oligofectamine组(250.7±14.57,506.09±19.14,711.79±19.91)24,48,72 h VEGF蛋白的分泌浓度均显著低于Control组(394.23±16.26,711.6±26.21,933.85±28.65)(P<0.05),且ASODN+Oligofectamine的抑制作用比ASODN强(P>0.05).结论:Oligofectamine介导的VEGF ASODN能抑制GBC-SD细胞VEGF,Flt-1及KDR mRNA表达和VEGF蛋白分泌.AIM： To investigate the effect of oligofectaminemediated vascular endothelial growth factor （VEGF） antisense oligodeoxynucleotide （ASODN） transfection on the mRNA expression of VEGF, fins-like tyrosine kinase-1 （Flt-1） and kinase insert domain-containing receptor （KDR） as well as VEGF protein excretion of gallbladder carcinoma GBC-SD cells in vitro. METHODS： Gallbladder carcinoma GBC-SD cells were transfected with VEGF ASODN and scrambled oligodeoxynucleotide （SODN） by Oligofectamine mediation. The mRNA expression of VEGF, Flt-1 and KDR in GBC-SD cells of each group were detected by semi-quantitive reverse transcription-polymerase chain reaction （RT- PCR） and the excretion of VEGF protein was measured by enzyme-linked imrnunosorbent assay （ELISA）. RESULTS： Semi-quantitive RT-PCR revealed that VEGF, Flt-1 and KDR mRNA expression in groups of ASODN （VEGF165：0.686 ± 0.033, 0.569 ± 0.049, 0.489 ± 0.036, 0.716 ± 0.017; VEGF165： 0.462 ± 0.046, 0.338 ± 0.034, 0.219 ± 0.022, 0.471 ± 0.038; Flt-1： 0.694 ± 0.019, 0.562 ± 0.045, 0.435 ± 0.042, 0.724 ± 0.026; KDR： 0.667 ±0.063, 0.490 ± 0.033, 0.301 ± 0.029, 0.665 ± 0.068） and ASODN ＋ Oligofectamine （VEGF165： 0.601 ± 0.021, 0.465 ± 0.042, 0.416 ± 0.023, 0.662 ± 0.035; VEGF121： 0.408 ± 0.014, 0.286 ± 0.019, 0.157 ± 0.021, 0.418 ± 0.037; Flt-1： 0.609 ± 0.018, 0.442 ± 0.049, 0.314 ± 0.015, 0.614 ± 0.029; KDR： 0.523 ± 0.048, 0.432 ± 0.027, 0.218 ± 0.036, 0.524 ± 0.037） were significantly inhibited 24, 48, 72 and 96 hours after transfection in comparison with those in the control group （P 〈 0.05）, and the inhibitory effect of ASODN ＋ Oligofectamine was stronger （P 〉 0.05）. ELISA results discovered that VEGF protein excretion was markedly decreased in the culture media of ASODN （281.26 ± 18.62, 526.44 ± 34.95, 791.13 ± 20.99） and ASODN ＋ Oligofectamine （250.7 ± 14.57, 506.09 ± 19.14, 711.79 ± 19.91） group （P 〈 0.05） as compared with that in the control

关 键 词：胆囊癌细胞GBC-SD 血管内皮生长因子 fms样酪氨酸激酶1 含激酶插入区受体 反义寡核苷酸 半定量RT-PCR 酶链免疫吸附试验 

分 类 号：R735.8[医药卫生—肿瘤]