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Abstract
BackgroundOur 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.MethodsmiR-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.ResultsThirteen 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.ConclusionThis 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.
Highlights
Our previous studies indicated that miR-200b inhibits the growth of androgen-independent prostate cancer (AIPC) cells
Identification of differentially‐expressed proteins (DEPs) by label free proteomics In this study, a total of 3792 proteins were identified in 4 samples by Liquid chromatography (LC)-ESI–MS/MS
In order to verify the regulation of the proteins at the protein level, PPP1R2, MARCKS, and GLIPR2 were randomly chosen for validation by Western blot analysis
Summary
Our previous studies indicated that miR-200b inhibits the growth of androgen-independent prostate cancer (AIPC) cells. We employed quantitative proteomics techniques to unravel the role of miR-200b in AIPC. Androgen deprivation therapy (ADT) is an effective therapeutic option for PCa. clinical outcomes of ADT in PCa patients have remained unsatisfactory because of castration resistance [2]. Clinical outcomes of ADT in PCa patients have remained unsatisfactory because of castration resistance [2] Small endogenous non-coding RNA (18–25 nt) are known as miRNAs. Accumulating evidence indicates that miRNAs play a critical role in AIPC. Previous studies have revealed that miR-200b are down-regulated in AIPC cells and that this phenomenon contributes to androgenindependent growth [3, 4]. Functional characterization of miRNAs depends strongly on identification of their specific mRNA binding partners because miRNAs can prevent protein expression by binding either the 3′UTR of
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