Screening and expression of specific SiRNA targeting IKKβ in human hypertrophic scar fibroblasts and construction of IKKβ-siRNA expression vector
AIM: To explore the effects of different siBNAs on the expression of IKKβ mRNA and to construct the specific IKKβ-siRNA vector of human hypertrophic scar fibroblasts. METHODS: Three IKKβ-siRNAs (L1, L2 and L3) were transcribed in vitro and then transferred to cultured human hypertrophic scar fibroblasts. BT-PCR was performed to evaluate the levels of IKKβ mRNA in the transferred cells. The specific IKKβ-siRNAs plasmid vector was constructed according to the screened IKKβ-siRNA siRNAs. RESULTS: The expression of IKKβ-siRNA mRNA of fibroblasts transferred with only L2 reduced significantly. Double enzyme digestion and sequence detection showed that the plasmid vector of siRNA specific for IKKβ was successfully constructed. The expression of IKKβ was significantly suppressed at both mRNA and protein levels after transfection. CONCLUSION: Specific IKKβ-siRNA of human hypertrophic scar fibroblasts can be screened out by RT-PCR. The IKKβ-siRNA expression vector of potential interference ability fur target tissue can be constructed. The expression of targeting IKKβ-siRNA of fibroblasts transferred reduced significantly.
NETL
NSTL
万方数据
维普
