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

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Author:
No author available
Journal Title:
WORLD CHINESE JOURNAL OF DIGESTOLOGY
Issue:
11
DOI:
10.3969/j.issn.1009-3079.2007.11.009
Key Word:
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

Abstract´╝Ü 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24,48,72,96h 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±0.019,0.1 57±0.021,0.418±0.037)、Flt-1 (ASODN组:0.694±0.01 9,0.562±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±0.027,0.21 8±0.036,0.524±0.037)mRNA的表达显著低于Contril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±1 8.62,526.44±34.95,791.1 3±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±1 6.26,711.6±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.

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