Suppression of Survivin expression by small interference RNA inducing apoptosis of keloid fibroblasts

Abstract

Objective To evaluate the effect of Survivin small interfering RNA (siRNA) silencing Survivin gene on the apoptosis rate and proliferation of fibroblasts in keloid. Methods The plasmid pSIREN-siRNA/Survivin was constructed and transfected into fibroblasts via LipofectamineTM 2000. The expression level of Survivin mRNA and protein was detected by using reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting after thtransfection. Apoptosis rate and cell cycle were measured by flow cytometry. Cellular proliferation was measured by methyl thiazol tetrazolium (MTT) assay. Results siRNA was transfected into fibroblasts effectively and its transfection efficiency was 67.0%. Survivin mRNA and protein were inhibited after transfection for 24 h and the inhibitory rate was 52. 2% and 54. 8%respectively (P ＜0. 05). The inhibitory rate was increased to 75.7% and 76.7% after transfection for 48 h. Amount of S phase cells was decreased, while that of G2/M phase increased. The apeptosis rate 24 and 48 h after transfection was 11.9% and 21.2% respectively, significantly higher than in negative and untreated groups. MTT asasy showed that keloid fibroblasts proliferation was inhibited after transfection of plasmid pSIREN-siRNA/Survivin. Conclusion siRNA silencing Survivin gene can effectively inhibit its mRNA and protein expression and fibroblast proliferation, and induce apoptosis. Key words: Keloid; RNA interference; Survivin gene; Gene therapy