VEGF-C基因转染对人乳腺癌MCF-7细胞VEGF-C表达的影响

Effect of VEGF-C Gene Transfection on Expression of VEGF-C in Human Breast Cancer MCF-7 Cell

目的探讨将真核表达载体pcDNA3.1-VEGF-C重组质粒转染入人乳腺癌MCF-7细胞后VEGF-C表达的变化。方法通过脂质体介导方法,将构建好含有VEGF-C的重组质粒转染入人乳腺癌MCF-7细胞中,新霉素(G418)筛选得到稳定转染细胞系,RT-PCR和Western blot方法检测稳定转染后细胞中VEGF-C mRNA和蛋白的表达。结果成功转染并获得稳定高表达VEGF-C的乳腺癌MCF-7细胞系,其转染组VEGF-C mRNA相对吸光度值(12.382±2.183)较空载组(6.039±1.950)显著上调(P<0.01)。Western blot检测转染组VEGF-C蛋白相对灰度值(0.971±0.186)较空载组(0.594±0.196)明显上调(P<0.05)。结论脂质体介导重组质粒pcDNA3.1-VEGF-C转染入人乳腺癌MCF-7细胞中可显著增加VEGF-C表达水平。

Objective To explore the effect of vascular endothelial growth factor-C （VEGF-C） gene transfection on the expression level of VEGF-C in human breast cancer MCF 7 cell. Methods The constructed VEGF-C gene eukaryotie expression vector was transfected into the human breast cancer MCF-7 cell by using lipofectamine transfection reagents, and the positive cell clones were obtained through G418 selection after transfection. The ex pressions of VEGF-C mRNA and protein were detected by RT-PCR and Western blot respectively. Results Follow- ing the transfection of the VEGF-C recombination plasmid, there were significant differences on the expression levels of VEGF-C mRNA and protein between peDNA3.1-VEGF-C transfeetion group and pcDNA3. 1 transfection group （12.382±2.183 vs 6.039±1.950, P〈0.01; 0.971±0.186 vs 0.594±0.196, P〈0.05）. Conclusion With the transfeetion of pcDNA3.1-VEGF-C vector by using the liposome, the expression levels of VEGF-C mRNA and protein rise up in breast cancer MCF-7 cell.