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目的利用RNA干扰(RNAi)技术,研究血管内皮生长因子(VEGF)编码基因的siRNA对人视网膜母细胞瘤(RB)细胞的影响,从而探讨通过抑制血管内皮生长因子在视网膜母细胞瘤组织中的表达来治疗视网膜母细胞瘤的机理。方法将VEGF编码基因特异的siRNA转染入视网膜母细胞瘤细胞,采用RT-PCR和Western-blot技术检测siRNA处理前后RB细胞VEGF基因表达变化。MTT法检测siRNA处理前后RB细胞生长的抑制情况。结果RT-PCR和Western-blot均显示VEGF特异性siRNA对视网膜母细胞瘤细胞血管内皮生长因子基因的表达有抑制作用,MTT法检测RB细胞生长的抑制情况显示,抑制率可达40%。结论VEGF特异性siRNA的特异性基因沉默作用对RB细胞VEGF基因的表达有抑制作用。
OBJECTIVE: To study the effect of siRNA encoding vascular endothelial growth factor (VEGF) gene on human retinoblastoma (RB) cells by RNA interference (RNAi) technology, and to explore the possible mechanism by inhibiting the expression of vascular endothelial growth factor in retinoblastoma tissue Expression to treat retinoblastoma mechanism. Methods VEGF gene encoding siRNA was transfected into retinoblastoma cells. The expression of VEGF gene in RB cells was detected by RT-PCR and Western-blot before and after siRNA treatment. The inhibition of RB cell growth was detected by MTT assay before and after siRNA treatment. Results Both RT-PCR and Western-blot showed that VEGF-specific siRNA inhibited the expression of vascular endothelial growth factor gene in retinoblastoma cells. The inhibition of RB cell growth by MTT assay showed that the inhibitory rate was up to 40%. Conclusion The specific gene silencing of VEGF-specific siRNA can inhibit the expression of VEGF gene in RB cells.