反义血管内皮生长因子对乳腺癌细胞生长的抑制作用  

The effect of VEGF_ (165) antisense RNA on human breast cancer cell

作　　者：李国栋[1] 宋燕[1] 刘力华[2] 段秀梅[2] 

机构地区：[1]吉林大学中日联谊医院基本外科,吉林长春130033 [2]吉林大学第一医院中心实验室

出　　处：《中国实验诊断学》2005年第2期187-189,共3页Chinese Journal of Laboratory Diagnosis

基　　金：吉林省科技厅基金资助课题(20010526)

摘　　要：目的　构建表达反义血管内皮生长因子165(VEGF165)的真核表达载体,观察其对人乳腺癌细胞MCF- 7的影响,探讨反义基因治疗乳腺癌的可行性。方法　应用基因重组技术,将人VEGF165cDNA反向克隆入pcDNA3真核表达载体中,构建VEGF165反义基因的真核表达载体,将此表达载体转染人乳腺癌细胞MCF -7,观察转染前后MCF 7的VEGF165表达及细胞生长周期。结果　成功构建出VEGF165反义RNA真核表达载体,反义质粒转染后MCF -7细胞VEGF165表达下降,G1期细胞增加,S期细胞减少,细胞增殖能力降低。结论　VEGF165反义RNA能够明显减少人乳腺癌细胞株MCF- 7内VEGF165的表达,抑制乳腺癌细胞的增殖,证明了反义基因干预治疗的有效性,为乳腺癌的基因治疗进行了有益探索。Objective To construct eukaryotic expression vector carrying antisense VEGF_ 165 and investigate whether the recombined plasmids could regulate the secretion level of VEGF protein and affect the growth cycle of the transfected human breast cancer cell line MCF-7. Methods VEGF_ 165 cDNA was amplified by reversetranscription PCR(RT-PCR) from MCF-7 cell, and was then inserted into the expression plasmid pcDNA3 to construct the recombined plasmids that encoding VEGF_ 165 cDNA in an antisense orientation. The MCF-7 were transfected with these plasmids. The VEGF expression of MCF-7 cells before or after transfection was detected by immunochemistry. The growth cycle was also detected. Results The eukaryotic expression vector pcDNA3/anti-VEGF165 was successfully constructed. Antisense VEGF gene decreased the level of VEGF protein in the transtected MCF-7 cells. The proliferation ability of MCF-7 cell was decreased after transfected, G1 phase cells were increased and S phase cells were decreased in cell cycle. Conclusion The successful construction of antisense VEGF eukaryotic expression vector is of significance for breast cancer specific antisense gene therapy. It may be applied to treat solid tumor in the future.

关 键 词：血管内皮生长因子 乳腺癌 反义基因 

分 类 号：R737.9[医药卫生—肿瘤]