血管内皮生长因子反义基因真核表达载体的构建及其对乳腺癌细胞的生长抑制作用

目的构建反义血管内皮生长因子(VEGF165)基因真核表达载体,分析该基因对乳腺癌细胞的生长抑制作用。方法将人VEGF165cDNA反向克隆至pcDNA3真核表达载体中,构建VEGF165反义基因的真核表达载体,转染人乳腺癌细胞MCF-7,观察转染前后MCF-7细胞的VEGF165表达及细胞生长周期。结果所构建的VEGF165反义基因真核表达载体转染MCF-7细胞后,VEGF165表达下降,细胞生存率下降,G1期细胞数量增加,S期细胞数量减少,细胞增殖能力降低。结论成功构建了VEGF165反义基因表达载体,该基因对乳腺癌细胞的生长具有明显的抑制作用。

Construction of Eukaryotic Expression Vector for Antisense Vascular Endothelial Growth Factor_(165) cDNA and Its Inhibitory Effect on Growth of Breast Cancer Cells

Objective To construct the eukaryotic expression vector for antisense vascular endothelial growth factor 165 (VEGF165) cDNA and study the inhibitory effect of antisense VEGF165 cDNA on the growth of breast cancer cells. Methods Amplify human VEGF165 cDNA from breast cancer MCF-7 cells by RT-PCR and inversely insert into eukaryotic expression vector pcDNA3. Transfect MCF-7 cells with the constructed recombinant plasmid pcDNA3 / anti-VEGF165 and observe the expression of VEGF165 and cell cycle before and after transfection. Results Sequencing result proved that recombinant plasmid pcDNA3 / anti-VEGF165 was constructed correctly. After transfection with the recombinant recombinant plasmid, both the expression level of VEGF165 as well as survival rate and proliferation ability of MCF-7 cells decreased, and the percentage of MCF-7 cells at G1 phase increased while that at S phase decreased. Conclusion The eukaryotic expression vector for antisense VEGF165 cDNA was successfully constructed, which inhibited the growth of breast cancer MCF-7 cells significantly.