【摘要】 目的探讨采用超声辐照联合超声造影剂介导含血管内皮生长因子受体-胸腺激酶(KDR-TK)基因的真核表达质粒载体转染血管内皮细胞的有效性,以及调控单纯疱疹病毒I型的胸腺激酶(HSV-TK)在血管内皮细胞中特异性杀伤作用。方法基因重组构建真核表达质粒载体pEGFP-KDR-TK,采用超声辐照联合超声造影剂SonoVue介导方式转染血管内皮细胞,MTT检测更昔洛韦(GCV)对KDR-TK/VEC(VEC)、原代人脐因管内皮细胞(HUVEC)、人肝癌细胞株(HepG2)的杀伤作用。结果GCV浓度为1、10、50和100μg/ml时,KDR-TK/VEC和HUVEC细胞抑制率分别为14.7%±11.0%vs5.2%±3.3%,P=0.412;44.9%±24.6%vs13.2%±1.7%,P=0.010;68.8%±7.0%vs26.2%±9.3%,P=0.001和73.2%±32.8%vs38.8%±16.8%,P=0.006。而HepG2细胞抑制率仅为6.1%±5.1%,9.4%±1.9%,13.2%±0.4%和14.7%±6.9%,与前两者比较,P均<0.05。结论超声辐照联合SonVue能有效地介导含KDR-TK真核表达质粒转染血管内皮细胞,可调控HSV-TK/GCV对血管内皮细胞的特异性杀伤作用。更多还原

【Abstract】 Objective To investigate the feasibility of the recombinant eukaryotic expressive plasmid containg kinase domain insert containing receptor-thymidine kinase (KDR-TK)transfering into vascular endothelial cells mediated by combination ultrasound irradiation with contrast agent.We tried to confirm the special killing efficiency of herpes simplex virsus-TK-ganciclovir(HSV-TK/GCV)against vascular endothelial cell mediated by combination of irradiation with contrast agent.Methods Eukaryotic expressive plasmid vector (pEGFP-KDR-TK) constructed by gene recombination was transfected into vascular endothelial cell mediated by combination ultrasound irradiation with contrast agent(SonoVue).The killing efficiency of GCV against KDR-TK/ VEC,human umbilical vascular endothelial cell(HUVEC) and hepatic carcinoma G2(HepG2) were measured with MTT.Results When the concentration of GCV were 1 μg/ml,10 μg/ml,50 μg/ml and 100 μg/ml,the cell inhibiting rate of KDR-TK /VEC and HUVEC were 14.7%±11.0% vs.5.2%±3.3%,P=0.412; 44.9%±24.6% vs 13.2%±1.7%,P=0.010; 68.8%±7.0% vs 26.2%±9.3%,P=0.001 and 73.2%±32.8% vs 38.8%±16.8%,P=0.006.On the other hand,the cell suppressive rates of HepG2 were 6.1%±5.1%,9.4%±1.9%,13.2%±0.4% and 14.7%±6.9%,compared with KDR-TK /VEC and HUVEC respectively,P<0.05.Conclusions Combination of ultrasoundirradiation with SonoVue could efficiently mediate transfection ofrecombinant eukaryotic expressive plasmid containing KDR-TK into vascular endothelial cell and regulated special killing efficiency of HSV-TK/GCV against vascular endothelial cell.更多还原