VEGF反义寡核苷酸对人胆囊癌细胞VEGF、Flt-1及KDR mRNA和蛋白水平表达的影响  

Effects of vascular endothelial growth factor (VEGF) anfisense oligodeoxynucleotide on mRNA and expression of VEGF, Fit-1, and kinase insert domain containing receptor and VEGF excretion in human gallbladder carcinoma cells

作　　者：方河清[1] 李海军[2] 刘颖斌[1] 王许安[1] 马孝明[1] 孔颖[1] 陈燕[1] 陈德琴[1] 翁伟宏[1] 张志平[1] KR Devkota 王建伟[1] 李江涛[1] 曹利平[1] 彭淑牖[1] 

机构地区：[1]浙江大学医学院附属第二医院外科,杭州310009 [2]新疆医科大学附属肿瘤医院肝胆外科

出　　处：《中华医学杂志》2007年第47期3329-3334,共6页National Medical Journal of China

基　　金：国家自然科学基金(30571831)

摘　　要：目的体外研究 Oligofectamine 介导的血管内皮生长因子(VEGF)反义寡核苷酸(ASODN)转染对人胆囊癌细胞 VEGF、Flt-1及 KDR 在 mRNA 和蛋白水平的表达及生长增殖和凋亡的影响。方法运用 Oligofectamine 介导的 VEGF ASODN 和 SODN 转染人胆囊癌细胞 GBC-SD,半定量 RT-PCR 检测各组细胞 VEGF、Flt-1及 KDR mRNA 转染前后不同时间的表达变化、ELISA 测定转染后各组细胞培养上清液 VEGF 蛋白浓度。结果 ASODN 组及 ASODN+Oligofectamine 组都能显著抑制 VEGF、Flt-1及 KDRmRNA 的表达,且 VEGF ASODN+Oligofectamine 的抑制作用比 ASODN 更强(P>0.05)。ELISA 测定结果显示 ASODN 组及 ASODN+Oligofectamine 组均抑制 VEGF 蛋白的分泌(P<0.05),且 ASODN+Oligofectamine 组的 VEGF 蛋白浓度低于 ASODN 组(P>0.05)。结论 VEGFASODN 能抑制人胆囊癌细胞 VEGF、Flt-1和 KDR 在 mRNA 水平的表达和 VEGF 蛋白的表达,从而促进 GBC-SD 细胞的凋亡;Oligofectamine 能明显增强 VEGF ASODN 的抑制效果。Objective To investigate the effects of vascular endothelial growth factor （VEGF） antisense oligodeoxynucleotide （ASODN） on the mRNA and protein expression of VEGF, Fit-1, and kinase insert domain containing receptor （KDR） and VEGF excretion in human gallbladder carcinoma cells. Methods Human gallbladder carcinoma cells of the line GBC-SD were cultured and transfected with VEGF ASODN and sense oligodeoxynucleotide （SODN） mediated by Oligofectamine. The toxicity of SODN and Oligofectamine to the GBC-SD cells was examined by MTT method. RT-PCR was used to detect the mRNA and expression of VEGF, Fit-1, and KDR, and ELISA was used to detect the protein expression of VEGF. Results MTT method showed that SODN and Oligofectamine were not toxic to the GBC-SD cells. The mRNA expression levels of VEGF, Fit-l, and KDR of the ASODN and ASODN ＋ Oligofectamine groups were all significantly lower than those of the control group （ all P 〈 0. 05 ）, and were the lowest 72 hours after transfection, and then gradually increased. ELISA showed that. there were not significant differences in the VEGF protein concentration in the supematant of the GBC-SD cells among the SODN, SODN ＋ Oligofectamine, and control groups （all P 〈 0. 05 ）, however, the VEGF protein concentration in the supematant of the GBC-SD cells of the ASDN and ASDN ＋ Oligofectamine groups were significantly lower than that of the control group （ both P 〈 0. 05 ）. Conclusion VEGF ASODN inhibits the mRNA and protein expression of VEGF, Fit-l, and KDR and VEGF excretion in human gallbladder carcinoma cells. Oligofectamine strengthens the effect of ASODN.

关 键 词：寡核苷酸 反义 内皮生长因子 GBC-SD细胞 Oligifeetamine 

分 类 号：R735.8[医药卫生—肿瘤]