The Role of miRNAs in the Progression of Prostate Cancer from Androgen-Dependent to Androgen-Independent Stages
Abstract
We used two genomic strategies for microRNA profiling, next generation sequencing and Locked Nucleic Acid miRNA microarrays, and verified the concordant changes by quantitative RT-PCR. miR-125b and members of miR-99 family (miR-99a, -99b, -100) were down-regulated in all three assays in the more advanced prostate cancer cell line C4-2 relative to the parental LNCaP cells. Similar decrease was seen in the transformed prostate cancer cell line WPE1-NB26 compared to its untransformed parent, RWPE1. Interestingly, all four miRNAs were also down-regulated in human prostate tumor tissue compared to normal prostate. Transfection of miR-99a, -99b or -100 inhibited the growth of prostate cancer cells and decreased the expression of prostate-specific antigen (PSA) indicating their potential role as tumor suppressors. To identify validated targets, we combined the computational prediction of potential targets, microarrays to detect mRNAs down-regulated by miRNAs and an analysis of the loading of mRNA on polyribosomes. We validated three direct targets of the miR-99 family: chromatin remodeling factors SMARCA5 and SMARCD1 and a kinase involved in signal transduction, mTOR and demonstrated that the expression of PSA is post-transcriptionally regulated by miR-99 family at least partially through repression of SMARCA5.
Document Details
- Document Type
- Technical Report
- Publication Date
- Sep 01, 2011
- Accession Number
- ADA554213
Entities
People
- Dandan Sun
Organizations
- University of Virginia