Label free quantitative proteomics reveals the role of miR-200b in androgen-independent prostate cancer cells

Abstract

Abstract Background Our previous studies indicated that miR-200b inhibits the growth of androgen-independent prostate cancer (AIPC) cells. In this study, we employed quantitative proteomics techniques to unravel the role of miR-200b in AIPC. Methods miR-200b was over-expressed or inhibited by transfection with miR-200b mimics or miR-200b inhibitor in PC3 cells. Total proteins were collected and the profiles of different groups were analyzed by label-free proteomics. PANTHER was applied to analyze biological processes, molecular functions and pathways of proteins regulated by miR-200b. miRBase was used to evaluate target genes of miR-200b in proteins regulated by miR-200b. Results Thirteen proteins were up-regulated in miR-200b mimics/mimics NC (negative miRNA control) and 14 proteins were down-regulated; 67 proteins were up-regulated in miR-200b inhibitor/inhibitor NC and 98 proteins were down-regulated. There were seven proteins which were both down-regulated by miR-200b mimics and up-regulated by miR-200b inhibitor, TM4SF1, YAP1, PPP1R2, MARCKS, RTN4, GLIPR2 and SUCLG1. Among these, TM4SF1, YAP1, PPP1R2, MARCKS, RTN4 were predicted as target genes of miR-200b by miRBase, while GLIPR2 and SUCLG1 were not. Conclusion This work identified several target genes of miR-200b by label free proteomics method, i.e., TM4SF1, YAP1, PPP1R2, MARCKS, RTN4, GLIPR2, and SUCLG1. The signaling pathways regulated by these proteins such as Hippo signaling may contribute to the phenotype resulting from miR-200b.

Keywords

• Prostate cancer
• Androgen-independent prostate cancer
• miR-200b
• Proteomics