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Synuclein-gamma suppression mediated by RNA interference inhibits the clonogenicity and invasiveness of MCF-7 cells

机译:RNA干扰介导的突触核蛋白γ抑制抑制MCF-7细胞的克隆形成性和侵袭性

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摘要

The aim of the present study was to investigate the effects of synuclein-gamma (SNCG) downregulation by RNA interference (RNAi) on the clonogenicity and invasiveness of MCF-7 breast cancer cells. This study used four pairs of SNCG-specific siRNAs which were designed and cloned into the pGPU6 plasmid for introduction into an MCF-7 cell line. The SNCG knockdown efficacies of the four siRNAs were compared using the reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemistry. The cells' clonogenic and invasive phenotypes were examined with clonogenic and Boyden chamber assays. In comparison with the non-specific siRNA and empty vector controls, all four SNCG siRNAs were observed to significantly inhibit SNCG expression at the mRNA and protein levels (F=481.06, P<0.001; F=147.42, P<0.0001). SNCG suppression mediated by RNAi successfully inhibited the clonogenicity (P=0.002) and invasiveness (P<0.001) of transfected MCF-7 cells. According to the results of the present study, we concluded that SNCG suppression mediated by RNAi significantly suppressed SNCG expression at the mRNA and protein levels, suggesting that SNCG suppression mediated by an RNAi strategy may become a novel approach for treating advanced breast cancer.
机译:本研究的目的是研究RNA干扰(RNAi)对突触核蛋白-γ(SNCG)下调对MCF-7乳腺癌细胞克隆形成和侵袭性的影响。这项研究使用了四对SNCG特异性siRNA,这些siRNA被设计并克隆到pGPU6质粒中,以引入MCF-7细胞系。使用逆转录聚合酶链反应(RT-PCR)和免疫细胞化学比较了四个siRNA的SNCG敲低效率。用克隆形成和博登室测定法检查细胞的克隆形成和侵入表型。与非特异性siRNA和空载体对照相比,观察到所有四个SNCG siRNA在mRNA和蛋白水平上均显着抑制SNCG表达(F = 481.06,P <0.001; F = 147.42,P <0.0001)。 RNAi介导的SNCG抑制成功抑制了转染的MCF-7细胞的克隆形成性(P = 0.002)和侵袭性(P <0.001)。根据本研究的结果,我们得出结论,RNAi介导的SNCG抑制显着抑制了mRNA和蛋白质水平上的SNCG表达,这表明RNAi介导的SNCG抑制可能成为治疗晚期乳腺癌的新方法。

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