利用RNAi逆转录病毒载体系统对人朊病毒蛋白表达的稳定抑制

朊病毒是引起可传染的致死性海绵状脑病的致病因子,细胞中正常的朊病毒蛋白(PrPC)在该疾病病程发展中起着必不可少的作用。同时,PrPC已被证明在胃癌、乳腺癌等癌症中发挥着保护癌细胞的作用。根据人源PrPC(HuPrPC)cDNA序列,本研究设计了4种19nt的siRNA,将其构建成RNAi逆转录病毒载体系统,进行了其对HuPrPC表达的抑制效应的分析,从中获得了能高效稳定抑制HuPrPC表达的3种靶向序列,其中si626(5′-GGTTGAGCAGATGTGTATC-3′)的抑制效果最为明显,其抑制效率可达85%以上。随后,利用筛选出的si292和si626的稳定干扰细胞系进行了细胞浸润性实验,结果发现,PrPC干扰细胞系细胞浸润能力显著下降。这为进一步研究朊病毒疾病的基因治疗、以PrPC为靶标进行PrPC相关癌症的辅助治疗研究奠定了一定的基础。

Stable inhibition of human prion protein through a retrovirus-based RNAi system

Prion leads to fatal transmissible spongiform encephalopathies. Cellular prion protein (PrPC) is necessary in prion disease. At present, it is demonstrated that PrPC plays a protective role in several carcinomas, such as gastric and breast cancer. We designed four 19-nt siRNAs according to cDNA sequence of human PrPC and constructed retrovirus-based RNAi vectors. We evaluated the inhibitive effect of these sequences on HuPrPC (human PrPC) and selected out three sequences with stable and efficient inhibition...