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Experimental Study on the Suppression of Human Nuclear Receptor hLRH-1 via a Vector-based RNA Interference
Authors:Shui-Liang WANG  Ji-Liang FU
Institution:a PLA Center for Laboratory Medicine, Fuzhou General Hospital, Fuzhou 350025, China
b Department of Medicine and Life Science, Tongji University, Shanghai 200092, China
Abstract: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 vector-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.
Keywords:RNA interference  nuclear receptor  hLRH-1  suppression of gene expression
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