Liang Bo, Wang Xin-Jun, Shen Pei-Hong, Li Xue-Yuan, Cheng Hong-Wei, Shan Qiao, Guo Kui-Yuan, Cao Yu-Wen, Fan Qing-Xia, Zheng Rui-Feng, Li Bei, Zhang Wei, Li Yan-Wei, Yang Kai
Departments of Neurosurgery, Zhengzhou, Henan 450052, P.R. China ;
Oncol Lett. 2013 Apr;5(4):1347-1352. doi: 10.3892/ol.2013.1192. Epub 2013 Feb 14.
The aim of the present study was to investigate the effects of synuclein-γ (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.
本研究旨在探讨RNA干扰（RNAi）下调突触核蛋白γ（SNCG）对MCF-7乳腺癌细胞克隆形成能力和侵袭能力的影响。本研究使用了四对SNCG特异性小干扰RNA（siRNA），将其设计并克隆到pGPU6质粒中，导入MCF-7细胞系。使用逆转录聚合酶链反应（RT-PCR）和免疫细胞化学方法比较了这四种siRNA对SNCG的敲低效果。通过克隆形成实验和Boyden小室实验检测细胞的克隆形成和侵袭表型。与非特异性siRNA和空载体对照相比，观察到所有四种SNCG siRNA均能在mRNA和蛋白质水平上显著抑制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细胞的克隆形成能力（P = 0.002）和侵袭能力（P < 0.001）。根据本研究结果，我们得出结论，RNAi介导的SNCG抑制在mRNA和蛋白质水平上显著抑制了SNCG表达，表明RNAi策略介导的SNCG抑制可能成为治疗晚期乳腺癌的新方法。
