血管内皮生长因子反义核酸对胰腺癌细胞增殖、凋亡和血管生成的影响  

Effect on tumor cell proliferation,apoptosis and angiogenesis by adenovirus mediated gene transfer of antisense vascular endothelial growth factor

作　　者：邵成浩[1] 胡先贵[1] 刘瑞[1] 张怡杰[1] 唐岩[1] 曹贵松[1] 

机构地区：[1]第二军医大学附属长海医院普外科,上海200433

出　　处：《中华实验外科杂志》2006年第1期18-20,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目(30200275)

摘　　要：目的探讨腺病毒介导的血管内皮生长因子(VEGF)反义核酸对胰腺癌细胞增殖、凋亡和血管生成的作用。方法构建反向插入VEGF165基因的复制缺陷型腺病毒载体(Ad-αVEGF)。18只裸鼠皮下接种人胰腺癌细胞株SW1990,随机分成3组(n=3),1周后瘤体内分别注射磷酸盐缓冲液(PBS,100μl,PBS对照组)、报告基因LacZ重组腺病毒(100μl,Ad-LacZ对照组)、反义VEGF重组腺病毒(100μl,Ad-αVEGF治疗组),隔日1次,共4次。1个月后处死动物。PCNA染色、TUNEL法和CD31染色观察反义VEGF165基因转染对胰腺癌细胞增殖、凋亡和血管生成的影响。结果Ad-αVEGF治疗组PCNA阳性表达率为(38.1±6.8)%,较LacZ组(89.6±4.3)%、PBS对照组(92.1±5.2)%明显降低(P<0.01),LacZ组与PBS对照组之间差异无统计学意义(P>0.05)。Ad-αVEGF治疗组细胞凋亡率(32.3±3.8)%,明显多于LacZ组(8.6±7.6)%和PBS对照组(9.9±4.2)%(P<0.01),LacZ组与PBS对照组之间差异无统计学意义(P>0.05)。Ad-αVEGF治疗组肿瘤微血管密度(12±3),明显少于LacZ组(26±5)和PBS对照组(25±4)(P<0.01),LacZ组与PBS对照组之间差异无统计学意义(P>0.05)。结论反义VEGF165基因转染可以抑制肿瘤细胞增殖,增加细胞凋亡,减少了肿瘤内微血管数量,从而抑制肿瘤生长。Objective To investigate the effect on tumor cell proliferation, apoptosis and angiogenesis by adenovirus mediated gene transfer of antisense vascular endothelial growth factor. Methods Adenoviral vector containing the cDNA for VEGF165 in an antisense orientation （Ad-αVEGF） was constructed, A nude mice xenograft model was established by subcutaneous injection of SW1990 cells, a human pancreatic cancer cell line. Eighteen nude mice were randomized into 3 groups： PBS control group, reporter gene Lac-Z adenovirus group and Ad-αVEGF treated group. All animals were treated once2 days with 4 times. Immunohistochemistral staining was used to investigatetumor angiogenesis, tumor cell proliferation and apoptosis after one month. Results Positive rate of the PCNA expression in Ad-αVEGF group （38.1 ± 6.8） % was significantly lower than that in Ad-LacZ group （92.1 ±5.2） % and PBS group （89.6 ± 4.3） % （P 〈 0.01）. Tumor cell apoptotic rate in Ad-αVEGF group （32.3 ± 3.8） % was significantly higher than that in Ad-LacZ group （8.6 ± 7.6） % and PBS group （9.9 ± 4.2） % （P 〈 0.01）. Microvessel density in Ad-αVEGF group （12 ± 3 ） was significantly lower than that in Ad-LacZ group （26 ± 5） and PBS group （25 ± 4） （P〈 0.01）. Conclusion Ad-α VEGF can significantly inhibit the tumor cell proliferation, increase the tumor cell apoptosis and reduce the amount of tumor micrevessels. The treatment of pancreatic cancer may be achieved by using an adenovirally mediated antisense- VEGF transfer.

关 键 词：血管内皮生长因子反义核酸 胰腺癌 细胞增殖 细胞凋亡 血管生成 微血管 肿瘤生长 

分 类 号：R735.9[医药卫生—肿瘤]