Effect of silencing SATB1 gene expression by RNA interference on invasive potential of human prostate carcinoma cells

Abstract

Objective To investigate the effect on invasion of human prostate carcinoma cells by applying RNA interference (RNAi) technique to silence special AT rich sequence binding protein-1 ( SATB1 ) gene.Methods The plasmid vector expressing SATB1 specific short hairpin RNA (shRNA) were constructed and transfected into human prostate carcinoma cell line DU145 by lipofectamine 2000.The pGPH1/GFP/Neo empty vector and untransfection group were used as negative control.Cells were collacted and assayed at 24,48,72 h after transfection.The SATB1 mRNA expression measured by reverse transcription-polymerase chain reaction (RT-PCR).Cell proliferation was assayed by methyl thiazol tetrazolium (MTT) method.The invasion ability of cells were examined by Transwell assay.Results The recombinant plasmid pGPH1/GFP/Neo-SATB1 expressing SATB1 shRNA had been constructed successfully.SATB1 mRNA expression was (53.0 ±4.0)％,(49.3 ±3.1 )％,(34.1 ±4.3)％,The rate of cell proliferation inhibited was ( 53.2 ± 5.8 ) ％,( 62.3 ± 4.0 ) ％,( 76.7 ± 3.0 ) ％ at 24,48,72 h after transfecion respectively,and the invasion ability was 34.2 ± 4.2,markedly weaken than that in the orther two groups ( P ＜ 0.05 ).Conclusion Silencing SATB1 gene can inhibit SATB1 gene expressions,suppress proliferation and invasion ability of human prostate carcinoma cell line DU145 by RNAi technology. Key words: Prostate cancer; Cell invasion; SATB1 gene