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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 inhibitory effect of siRNA targeting vascular endothelial growth factor (VEGF) gene on breast cancer MCF-7 cells. Methods: Four small interfering RNAs (siRNAs) targeted to VEGF were designed, including symmetric siRNA (siRNA21 / 21, siRNA23 / 23) and asymmetric siRNA (aiRNA; aiRNA21 / 23, aiRNA19 / 21) . After siRNA transfection into MCF-7 cells, the proliferation and apoptosis of MCF-7 cells were detected by MTT and flow cytometry. The expression of VEGF gene and protein in MCF-7 cells was detected by RT-PCR and ELISA. Results: Compared with the control group, all four siRNAs, aiRNA21 / 23, siRNA23 / 23, aiRNA19 / 21 and siRNA21 / 21 could effectively inhibit the expression of VEGF mRNA [(71.4 ± 5.01)% and (40.0 ± 3.11)%, (37.2 ± 2.79)%, (11.1 ± 0.99)% vs (2.4 ± 0.11)%, P <0.01] and inhibited the proliferation of MCF-7 cells [(44.7 ± 5.38)%, (38.5 ± 5.67)%, (33.6 ± 2.18)%, (33.1 ± 3.18)% vs (2.2 ± 0.28)% respectively, P <0.01]. The inhibition effect of asymmetric aiRNA21 / 23 was the most obvious. Compared with the control group, aiRNA21 / 23 significantly promoted the apoptosis of MCF-7 cells [(49.9 ± 4.02)% vs (4.7 ± 0.91)%, P <0.01]. Conclusion: siRNA targeting VEGF gene can inhibit the proliferation of MCF-7 cells and promote apoptosis, especially the most obvious effect of asymmetric siRNA.