CAJ | 학술논문

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.
위탐토경RNA간섭법유도인핵수체hLRH-1적표체억제적가행성,통과설계병구건능표체파향인핵수체hLRH-1기인적siRNAs적간섭재체pShLRH-1.1화pShLRH-1.2,경지질체개도법전염인간암세포BEL-7402,RT-PCR법감정hLRH-1기인적표체억제효응,동시이동양방법분석초린산법니지합성매기인적표체정황.순시전염후분석결과표명,소구건적간섭재체pshLRH-1.1화pShLRH-1.2균능재세포수평유효유도hLRH-1기인적표체억제,억제솔고체약80%;여미전염화공재체전염대조조상비,hLRH-1기인표체수억적세포중초린산법니지합성매기인적표체정명현상조,제시hLRH-1가능재초린산법니지합성매기인적표체중기부조작용.
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 lipofectaminTM. RT-PCR was carried out to examine the inhibition ratio of hLRH-1 expression. The same method was also applied to analyze the expression of farnesyl pyrophosphate synthetase (FPPS) 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.