Inhibition of insulin receptor expression in β cells by siRNA

AIM: To inhibit the expression of insulin receptor(IR) gene in mouse insulinoma β TC-3 cells by siRNA.METHODS: Four pairs of siRNA targeting IR gene,named siRNA-1,siRNA-2,siRNA-3,siRNA-4,and one pair non-specific siRNA(control group) were chemically synthesized and then transfected into β TC-3 cells using cation liposome transfection reagent.The efficacies of transfection were observed with fluorescence microscope,and the expression levels of IR mRNA were detected by reverse transcriptase-polymerase chain reaction after transfection for 24 h.RESULTS: ① The transfection efficacies with 50,100,150,200 nmol/L siRNA into β TC-3 cells were(73.1±4.1)%,(92.9±3.4)%,(90.8±4.0)%,(93.9±2.8)%,respectively.Finally 100 nmol/L was chosen.② After transfection with 100 nmol/L siRNAs for 24 h,the expression of IR mRNA in β TC-3 cells was decreased in all 4 groups compared with control group;among them,the effect was the most obvious in siRNA-2 group,in which the expression was decreased by 70.5%.CONCLUSION: The siRNAs targeting IR mRNA can effectively inhibit IR expression in vitro,which offers an experimental basis for further researches on IR in β cells.