Inhibitory Effect of siRNAs on the Expression of Bladder Cancer-Associated Protein in HeLa Cell Line

Objective: To construct cell lines with silenced bladder cancer-associated protein(BLCAP) with novel designed small interfering RNAs(siRNAs).Methods: Three specific siRNAs targeting BLCAP gene were designed and cloned separately,and were named as pRNA-U6.1-B1,B2,B3-siRNAs.The inhibitory effect and time-efficiency of each siRNA were investigated,and the expression levels of mRNA and protein of the target genes were detected with RT-PCR and Western blot at 12,24 and 48 h after transfection with the siRNAs.Results: Each siRNA led to a downregulation of the expression of target genes(P0.05),but the degree of downregulation was different.No distinct changes were found in the group transfected with liposomal transfection reagent without siRNAs.The results indicated that pRNA-U6.1-B2-siRNA was the most effective siRNA.Transfection with pRNA-U6.1-B2-siRNA induced a marked decrease of corresponding BLCAP mRNA and protein levels at different time points(P0.05) when compared with the control group.Conclusion: Inhibitory effect varies with siRNAs targeting different sequences of the same gene.Transfection with either siRNAs induces a specific and efficient downregulation of BLCAP gene in HeLa cells positive for HPV16 that lasts for 48 hours.