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pVVP3和pVHN核酸疫苗的构建、表达及对肿瘤细胞的作用
引用本文:米志强,金宁一,龚伟,薛立娟,连海,解丽华,李萍.pVVP3和pVHN核酸疫苗的构建、表达及对肿瘤细胞的作用[J].中国生物化学与分子生物学报,2003,19(6):704-708.
作者姓名:米志强  金宁一  龚伟  薛立娟  连海  解丽华  李萍
作者单位:解放军军需大学,解放军基因工程重点实验室,长春,130062
基金项目:国家自然科学基金重大项目 (No .3 9893 2 90 4 3 ),国家“973”项目 (No .G1990 1190 2 )资助~~
摘    要:用pVAX1载体构建抗肿瘤核酸疫苗 .将鸡贫血病病毒 (CAV)的VP3基因和鸡新城疫病毒(NDV)HN基因插入载体pVAX1的多克隆位点 ,构建了 2个重组基因疫苗pVVP3和pVHN .转染OS732细胞 ,Western印迹及血凝试验检测HN基因表达状况及表达产物的活性 .RT PCR、DNALadder、TUNNEL染色检测VP3基因的表达及表达产物诱导OS732细胞凋亡作用 .酶切鉴定证实成功地构建了两个核酸疫苗 ,并能在真核细胞内表达 .含HN的核酸疫苗pVHN表达后具有血凝活性并致肿瘤细胞表面唾液酸含量降低 (P <0 0 5 ) .pVVP3的转染能导致OS732细胞凋亡 ,凋亡率可达87% .试验结果表明 ,pVVP3和pVHN两个核酸疫苗体外应用后能诱导肿瘤细胞凋亡并显著降低肿瘤细胞表面唾液酸含量

关 键 词:鸡新城疫病毒HN基因  鸡贫血病病毒VP3基因  抗肿瘤  
收稿时间:2003-12-20
修稿时间:2002年11月29

Construction and Expression of the Nucleic Acid Vaccine pVVP3 and pVHN and Its Effect on Tumor Cell
MI Zhi qiang,JIN Ning yi ,GONG Wei,XUE Li\|juan,LIAN Hai,XIE Li hua,LI Ping.Construction and Expression of the Nucleic Acid Vaccine pVVP3 and pVHN and Its Effect on Tumor Cell[J].Chinese Journal of Biochemistry and Molecular Biology,2003,19(6):704-708.
Authors:MI Zhi qiang  JIN Ning yi  GONG Wei  XUE Li\|juan  LIAN Hai  XIE Li hua  LI Ping
Institution:MI Zhi qiang,JIN Ning yi *,GONG Wei,XUE Li\|juan,LIAN Hai,XIE Li hua,LI Ping
Abstract:Nucleic acid vaccine pVVP3 and pVHN were constructed with HN gene from NDV and VP3 gene from CAV, respectively, using pVAX1 vector. pVVP3 and pVHN were transfected into OS732 tumor cell. HN gene expression was detected by Western blotting and hemagglutination test. The content of OS732 cell surface sialic acid was measured. The influence of VP3 gene on OS732 was investigated by TUNNEL assay and DNA Ladder electrophoresis. Enzyme restriction test indicated that pVVP3 and pVHN were correctly constructed. Apoptosis was induced by pVVP3 on OS732 tumor cell with the rate of 87%. The HN expression could reduce the sialic acid content of OS732. The result indicated that the nucleic acid vaccine pVVP3 and pVHN had potential antitumor effect.
Keywords:Newcastle disease virus HN gene  chicken anemia virus VP3 gene  antitumor
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