摘要
目的 :构建并鉴定反向插入 VEGF1 65c DNA的重组腺病毒 ,探讨腺病毒介导的 VEGF1 65反义核酸治疗胰腺癌的可行性。 方法 :应用基因重组技术 ,将 5 13bp的人 VEGF1 65c DNA反向克隆入腺病毒粘粒载体 p Ax CAwt。重组粘粒与腺病毒DNA- TPC混合后以磷酸钙共沉淀法转染 2 93细胞制备重组腺病毒。建立胰腺癌裸鼠皮下种植瘤模型 ,瘤体内直接注射重组腺病毒 ,观察肿瘤的生长及血管生成情况。结果 :成功构建了反向插入 VEGF1 65c DNA的腺病毒粘粒载体 p Ax CAwt-αVEGF并制备出高滴度的重组腺病毒。实验第 5周时肿瘤生长速度 Ad-αVEGF治疗组比 PBS和 Ad- L ac Z对照组均明显减慢(P<0 .0 1) ;肿瘤微血管密度 Ad- αVEGF治疗组比两个对照组均明显减少 (P<0 .0 1)。结论 :腺病毒介导的 VEGF1 65反义核酸可以抑制胰腺癌的血管生成和肿瘤的生长 ,为抗血管生成的基因治疗提供了实验依据。
Objective: To construct a replication deficient recombinant adenoviral vector containing the VEGF 165 cDNA in an antisense orientation and to study its feasibility in treatment of pancreatic cancer. Methods: The VEGF 165 cDNA was inserted into adenovirus cosmid vector pAxCAwt in an antisense orientation. The replication deficient recombinant adenovirus was generated by the method of calcium phosphate co-precipitation from the homologous recombination of the cosmid VEGF and Ad5-TPC DNA. Recombinant adenoviruses were propagated and titrated in 293 cells and purified by cesium chloride density purification. Intratumoral treatment with recombinant adenoviruses was repeated every other day for 4 times. The angiogenesis and tumor growth were investigated. Results:PAxCAwt-αVEGF adenovirus cosmid vector was successfully constructed and high titer recombinant adenovirus were generated.The growth rate and microvessel density in tumor were markedly lower in Ad-αVEGF group than those in control group. Conclusion: The recombinant adenovirus is successfully generated. The angiogenesis and tumor growth of pancreatic carcinoma are effectively inhibited by the recombinant adenovirus.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2002年第9期995-998,共4页
Academic Journal of Second Military Medical University
关键词
血管内皮
内皮生长因子
腺病毒科
胰腺肿瘤
基因治疗
endothelium,vascular
endothelial growth factors
adenoviridae
pancreatic neoplasms
gene therapy
