基于载体的RNA干涉介导人核受体hLRH-1的表达抑制实验研究

为探讨经RNA干涉法诱导人核受体hLRH-1的表达抑制的可行性，通过设计并构建能表达靶向人核受体hLRH-1基因的siRNAs的干涉载体pShLRH-1．1和pShLRH-1．2，经脂质体介导法转染人肝癌细胞BEL-7402，RT-PCR法鉴定hLRH-1基因的表达抑制效应，同时以同样方法分析焦磷酸法呢酯合成酶基因的表达情况。瞬时转染后分析结果表明，所构建的干涉载体pShLRH-1．1和pShLRH-1．2均能在细胞水平有效诱导hLRH-1基因的表达抑制，抑制率高达约80％；与未转染和空载体转染对照组相比，hLRH-1基因表达受抑的细胞中焦磷酸法呢酯合成酶基因的表达呈明显上调，提示hLRH-1可能在焦磷酸法呢酯合成酶基因的表达中起负调作用。

Experimental Study on the Suppression of Human Nuclear Receptor hLRH-1 via a Vector-based RNA Interference

To explore the inhibitions of human nuclear receptor hLRH-1 via RNA interference,siRNAs expressing vectors pShLRH-1.1 and pShLRH-1.2,and targeting hLRH-1 were designed and constructed.The recombinants were introduced into hepatocellular carcinoma cells,BEL-7402,mediated by lipofectamin TMRT-PCR was carried out to examine the inhibition ratio of hLRH-1 expression.The same method was also applied to analyze the expression of famesyl pyrophosphate synthetase (FTPS) gene.Our results demonstrated that after transient transfection,both pShLRH-1.1 and pShLRH-1.2 could trigger the efficient inhibition of hLRH-1 in cultured cells,BEL-7402.The inhibition ratios were up to 80%.By comparing with non-transfection and vec-tor-transfection control,the expression of FPPS in cells with inhibition of hLRH-1 was up-regulated significantly.Thus,the inhibition of expression of hLRH-1 in cultured cells was achieved via RNA interference in this study.Our results also suggested that hLRH-1 acts as a negative regulator in FPPS expression.