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G4PAMAM/VEGFASODN抗乳腺癌血管生成的体外实验研究 被引量:2
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作者 赵新汉 李丽 +2 位作者 王志宇 张灵小 安改丽 《浙江大学学报(医学版)》 CAS CSCD 2008年第6期612-621,共10页
目的:探讨G4PAMAM/VEGFASODN复合物体外对乳腺癌细胞MDA-MB-231VEGF、VEGF mRNA表达的影响及血管内皮细胞生长的抑制作用。方法:用透射电镜观测G4PAMAM/VEGFASODN的粒径,在不同的酸碱度下观察G4PAMAM/VEGFASODN的稳定性;将G4PAMAM/VEGFA... 目的:探讨G4PAMAM/VEGFASODN复合物体外对乳腺癌细胞MDA-MB-231VEGF、VEGF mRNA表达的影响及血管内皮细胞生长的抑制作用。方法:用透射电镜观测G4PAMAM/VEGFASODN的粒径,在不同的酸碱度下观察G4PAMAM/VEGFASODN的稳定性;将G4PAMAM/VEGFASODN进行体外转染,流式细胞仪测定其转染率,激光共聚焦检测转染阳性细胞,MTT法检测转染后细胞的存活率;免疫组化检测VEGF蛋白的表达,RT-PCR检测VEGFmRNA的表达,MTT法测定此复合物对血管内皮细胞生长的抑制作用。结果:G4PAMAM/VEGFASODN的直径约10nm,呈较均匀的网状排列;酸碱度在5到10的范围内具有较高的稳定性,不同电荷比的G4PAMAM/VEGFASODN均没有出现被解离的现象;48h时G4PAMAM/VEGFASODN组在1∶40的条件下转染率明显高于脂质体组;各组转染物均对细胞无明显毒性;VEGF蛋白在G4PAMAM/VEGFASODN转染后的MDA-MB-231细胞胞浆中的染色强度明显减弱,阳性表达率显著降低,VEGF mRNA的表达也受到有效的抑制。结论:G4PAMAM/VEGFASODN复合物稳定性好、转染率高,是一种有良好应用前景的新型纳米载体基因转移系统;G4PAMAM/VEGFASODN复合物能够高效特异地抑制目的基因VEGF的表达,为进一步进行体内动物实验提供了依据。 展开更多
关键词 乳腺肿瘤/药物疗法 基因疗法 g4pamam g4pamam/VEGFASODN 乳腺癌 VEGF VEGF mRNA 血管内皮细胞
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Anti-angiogenesis effect of generation 4 polyamidoamine/vascular endothelial growth factor antisense oligodeoxynucleotide on breast cancer in vitro 被引量:4
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作者 Shan-zhi GU Xin-han ZHAO +4 位作者 Ling-xiao ZHANG Li LI Zhi-yu WANG Min MENG Gai-li AN 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2009年第3期159-167,共9页
Objective:To study the effects of the generation 4 polyamidoamine/vascular endothelial growth factor antisense oligodeoxynucleotide(G4PAMAM/VEGFASODN) compound on the expressions of vascular endothelial growth factor(... Objective:To study the effects of the generation 4 polyamidoamine/vascular endothelial growth factor antisense oligodeoxynucleotide(G4PAMAM/VEGFASODN) compound on the expressions of vascular endothelial growth factor(VEGF) and its mRNA of breast cancer cells and on the inhibition of vascular endothelial cells. Methods:We examined the morphology of G4PAMAM/VEGFASODN compound and its pH stability,in vitro transfection efficiency and toxicity,and the expressions of VEGF and its mRNA. Methyl thiazolyl tetrazolium assay was used to detect the inhibitory function of the compound on vascular endothelial cells. Results:The compound was about 10 nm in diameter and was homogeneously netlike. From pH 5 to 10,it showed quite a buffered ability. The 48-h transfection rate in the charge ratio of 1:40 was 98.76%,significantly higher than that of the liposome group(P<0.05). None of the transfection products showed obvious toxicity on the cells. The expressions of both VEGF protein and its mRNA after G4PAMAM/VEGFASODN transfection decreased markedly. Conclusion:With a low toxicity,high safety,and high transfection rate,G4PAMAM/VEGFASODN could be a promising gene vector. Specifically,it inhibits VEGF gene expression efficiently,laying a basis for further in vivo animal studies. 展开更多
关键词 Breast cancer Vascular endothelial growth factor (VEGF) VEGF mRNA Generation 4 polyamidoamine(g4pamam Generation 4 polyamidoamine/vascular endothelial growth factor antisense oligodeoxynucleotide(g4pamam/VEGFASODN) Vascular endothelial cell
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