摘要
目的研究RNA干扰技术沉默血管内皮生长因子-C(vascular endothelial growth factor-C,VEGF-C)在人肺鳞癌SK-MES-1细胞中的表达情况,为肺癌的RNAi基因治疗提供实验依据。方法设计三组siRNAs(siRNA1、siRNA2、siRNA3),确认VEGF-C-siRNA,构建pGenesil.1-VEGF-C的干涉载体,稳定转染入肺鳞癌细胞株SK-MES-1,通过RT-PCR和Western-blot技术检测VEGF-C在细胞中的表达。结果 siR-NA1、siRNA2、siRNA3组均抑制SK-MES-1细胞增殖,其中siRNA1转染对SK-MES-1细胞VEGF-C增殖有明显抑制作用,其抑制率为72.45%,明显高于空白对照组、空脂质体组、阴性对照siRNA组(P<0.01);siR-NA-VEGF-C组细胞内VEGF-C蛋白表达明显低于空白对照组、空脂质体组,对VEGF-C蛋白表达下调率分别为54%、65%、6%,差异具有显著统计学意义(P<0.05)。结论 RNA干扰技术能够明显抑制SK-MES-1细胞中VEGF-C的表达,从而抑制肺癌发生淋巴转移。
Objective To investigate the inhibitation effect of VEGF-C expression in SK-MES-1 cells by RNA interfering.Methods Three small interfering RNAs（siRNAa,siRNAb,siRNAc）were prepared and designed VEGF-C-siRNA.The constructed pGenesil.1-VEGF-C vectors were stably transfected into SK-MES-1 cell line.Then the expression of VEGF-C was detected in stable transfection SK-MES-1 cell line by RT-PCR and Western-blot.Results RT-PCR results showed that siRNA1,siRNA2,siRNA3 could inhibit the growth of SK-MES-1 cells,among which siRNA1 was the most significant,with an inhibition rate of 72.45%,significantly higher than blank control group,empty plasmid group or siRNA negative group（P0.01）.The VEGF-C protein expression of siRNA-VEGF-C cells significantly lower than blank control group,empty plasmid group,down-regulated expression of VEGF-C protein was 54%,65%,6% respectively,there was significant statistical significance（P0.05）.Conclusion RNAi technology could silence the expression of VEGF-C in SK-MES-1 cells,Which suggested that the technology may be one of the effective methods for inhibiting lymphmetastasis in lung squre cancer.
出处
《中华肺部疾病杂志（电子版）》
CAS
2011年第1期22-25,共4页
Chinese Journal of Lung Diseases(Electronic Edition)
