摘要
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。
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 MTr 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/G~ 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 ceils proliferation inhibited and cells apoptosis increased.
出处
《广西医学》
CAS
2012年第3期267-271,共5页
Guangxi Medical Journal
基金
广西自然科学基金(桂科自0991230)
关键词
乳腺癌
血管内皮生长因子C
RNA干扰
慢病毒载体
细胞周期
Breast cancer
Vascular endothelial growth factor-C
RNA interference
Lentivirus vector
MCF-7
Cell cycl
