siRNA干扰乳腺癌MCF-7细胞VEGF-C表达及对细胞增殖影响的研究

目的研究慢病毒载体介导的siRNA对乳腺癌MCF-7细胞系血管内皮生长因子(VEGF-C)表达的敲减作用及对乳腺癌增殖和凋亡的影响。方法构建慢病毒VEGF-C/siRNA载体,转染乳腺癌MCF-7细胞,观察其转染效率,采用实时定量PCR检测MCF-7细胞在转染前后VEGF-C的mRNA表达,计算VEGF-C敲减率,采用四甲基偶氮唑蓝(MTT)法检测细胞增殖变化,流式细胞仪检测细胞周期及凋亡。结果慢病毒VEGF-C/siRNA转染效率超过80%,VEGF-C的mRNA表达敲减率达50%,MTT结果提示慢病毒VEGF-C/siRNA能有效抑制细胞增殖,流式细胞学实验结果提示VEGF-C/siRNA干扰后S期细胞明显减少,G0/G1期细胞显著增加,细胞凋亡增加。结论慢病毒VEGF-C/siRNA转染率高,能有效敲减VEGF-C的mRNA表达,使细胞增殖受到抑制,凋亡增加。

Down-regulation of VEGF-C Expression in MCF-7 Breast Cancer Cells With Lentivirus-mediated Small Interfering RNA Vectors and Its Effect on Cell Proliferation

Objective To study the knockdown of vascular endothelial growth factor-C(VEGF-C) expression of MCF-7 breast cancer cells interfered with lentivirus-mediated small interfering RNA vectors and its effect on cell proliferation and apoptosis.Methods Lentivirus-mediated small interfering RNA vectors were constructed and transducted into MCF-7 breast cancer cells,real-time PCR was performed to detecte the mRNA expression of VEGF-C.The proportion of transfected cells and the knock-down rate of VEGF-C were calculated.Proliferation of cells was measured by MTT assay.The apoptosis and cell cycle was analyzed by flow cytomotry(FCM).Results The transfection rate of MCF-7 cells interfered with lentivirus-mediated small interfering RNA was more than 80%,mRNA expression of VEGF-C decreased by nearly 50%,proliferation of the VEGF-C/SiRNA cells was inhibited.The result of FCM showed that less cells in S phase and more cells in G0/G1 phase after VEGF-C/siRNA interference than the controls,and apoptosis rate of MCF-7 cells increased significantly.Conclusion The transfection rate is high with lentivirus-mediated small interfering RNA vectors,VEGF-C gene expression is knocked down efficiently,which makes cells proliferation inhibited and cells apoptosis increased.