靶向VEGF的不对称siRNA抑制乳腺癌MCF-7细胞的增殖

Asymmetric siRNA targeting VEGF inhibits proliferation of breast cancer MCF-7 cells

目的:探讨靶向血管内皮生长因子(vascular endothelial growth factor,VEGF)基因的siRNA对乳腺癌MCF-7细胞的抑制效果。方法:设计靶向VEGF的4种小干扰RNA(small interfering RNA,siRNA),包括对称siRNA(siRNA21/21,siRNA23/23)与不对称siRNA(asymmetric siRNA,aiRNA;aiRNA21/23,aiRNA19/21)。siRNA转染入MCF-7细胞后,MTT及流式细胞术检测MCF-7细胞增殖及凋亡情况,RT-PCR、ELISA法检测MCF-7细胞中VEGF基因和蛋白的表达。结果:与对照组相比,aiRNA21/23、siRNA23/23、aiRNA19/21、siRNA21/21这4种siRNA都能有效抑制VEGF mRNA的表达[(71.4±5.01)%、(40.0±3.11)%、(37.2±2.79)%、(11.1±0.99)%vs(2.4±0.11)%,P<0.01],且抑制MCF-7细胞的增殖[(44.7±5.38)%、(38.5±5.67)%、(33.6±2.18)%、(33.1±3.18)%vs(2.2±0.28)%,P<0.01],其中以不对称aiRNA21/23的抑制效果最明显。与对照组比较,aiRNA21/23显著促进MCF-7细胞的凋亡[(49.9±4.02)%vs(4.7±0.91)%,P<0.01]。结论:靶向VEGF基因的siRNA可抑制MCF-7细胞的增殖、促进细胞凋亡,尤以不对称siRNA的效果最明显。

Objective： To investigate the inhibitiory effect of siRNA targeting vascular endothelial growth factor（VEGF） gene on proliferation of breast cancer MCF-7 cells.Methods： Small interfering RNA（siRNA） targeting VEGF,including 2 siRNAs（siRNA21 /21,siRNA23 /23） and 2 asymmetric siRNAs（aiRNA）（aiRNA19 /21,aiRNA21 /23） were designed and synthesized.The siRNA was transfected into MCF-7 cells,and cell proliferation was detected by MTT method,and the apoptosis was analyzed by flow cytometry.VEGF protein and mRNA expression levels of MCF-7 cells were detected by ELISA and RT-PCR.Results： Compared with the control group,the 4 siRNAs effectively inhibited VEGF mRNA expression（[71.4 ±5.01]%,[40.0 ±3.11]%,[37.2 ±2.79]%,[11.1 ±0.99]% vs[2.4 ±0.11]%,P 0.05）,and the proliferation of MCF-7 cells（[44.7 ± 5.38]%,[38.5 ± 5.67]%,[33.6 ± 2.18]%,[33.1 ± 3.18]%,[2.2 ± 0.28]%,P 0.05）,with aiRNA21 /23 of the best RNA interference effect.Compared with the control group,the apoptotic rate was significantly increased（[49.9 ± 4.02]% vs [4.7 ± 0.91]%,P 0.05）.Conclusion： siRNA targeting VEGF can inhibit proliferation and induce apoptosis of MCF-7 cells,with asymmetric siRNA showing best effects.