摘要
目的利用RNA干扰技术在视网膜母细胞瘤(retinoblastoma,RB)细胞内诱导RNA干扰(RNAi),抑制血管内皮生长因子(vascular endothelial growth factor,VEGF)基因表达,在体外细胞水平探讨RNAi对视网膜母细胞瘤治疗的可行性。方法构建靶向VEGF基因的siRNA(VEGF-siRNA),转导入RB细胞,在瘤细胞内诱导RNAi,采用RT-PCR、细胞增殖抑制实验、Northern杂交等技术检测siRNA处理前后RB细胞增殖及VEGF基因表达变化。结果成功构建了两种抗VEGF基因的shRNA表达载体,RT-PCR发现VEGF-siRNA在RB细胞系中,能明显抑制VEGF基因的表达,抑制率为72%;对RB细胞生长抑制率达47%;VEGF基因的mRNA表达降低了78%。结论VEGF-siRNA在体外能明显抑制了视网膜母细胞瘤细胞增殖和VEGF基因的表达。
Objective To investigate the feasibility of retinoblastoma （RB） gene therapy in which RNAi targeting was utilized to inhibit vascular endothelial growth factor （VEGF） gene expression in retinoblastoma cells. Methods Small interfering RNA （siRNA） homologous to the VEGF gene （VEGF-siRNA） was recombined with a pTZU6＋1 vector, then transfected into retinoblastoma cells to induce RNAi, and then alterations in VEGF gene expression and tumor cell proliferation in both the VEGF-siRNA treated groups and control groups were detected by MTT assay, Northern Blot and RT- PCR. Results siRNA can effectively inhibit the expression of VEGF in RB; the inhibition rate was 72%. The expression of VEGF was clearly inhibited by siRNA. The inhibition rate of cell proliferation was 47%. The mRNA level of VEGF decreased by 78%. Conclusion VEGF-siRNA effectively inhibited VEGF gene expression and retinoblastoma cell proliferation in vitro .
出处
《眼视光学杂志》
2007年第6期391-394,共4页
Chinese Journal of Optometry & Ophthalmology
