To identify oncogenic miRs in PC, miRs were screened by a commercial miR microarray service using normal prostate epithelial cell (RWPE-1) and PC cell lines (PC-3 and LNCaP).
We also used normal prostate and prostate cancer cell lines (RWPE-1, PC-3, DU-145) and performed in vitro functional analysis.
These data were compared with those obtained using normal primary fibroblast (C3PV), normal immortalized prostate epithelial cells (RWPE1) and normal non-tumorigenic breast epithelial cells (MCF 10A).
Calibration was performed using cDNA samples from normal prostate primary cells.
E-cadherin expression was first examined using immunofluorescence in 50 normal prostate tissues and in a cohort of 414 prostate cancer patients.
Although immortalized, RWPE-1 cells have retained properties exhibited by normal prostate epithelium in vivo.
In prostate cancer, developmentally important CGI genes silenced by PRC2 in normal prostate cells acquire DNA methylation concomitant with loss of their PRC2 marks 43.
Our previous studies also suggested that Id4 is regulated by androgens in normal prostate epithelial cells [ 32] and in androgen-sensitive prostate cancer cell line LNCaP [ 33].
These assays were then used to analyze 8 normal prostate tissues and 36 prostate tumors.
For this analysis we used 54 previously characterized normal prostate tissue samples [ 18, 19] and a real-time qRT-PCR protocol developed for prostate samples [ 20- 22].
