Analysis of gene expression of PCA3 transcript in different cell lines and its targeted knockdown by siPCA3 in PCa cells. (A) RNA expression of PCA3 was quantified by qRT-PCR in different prostate (LNCaP, PrEC, RWPE-1, DU145, and PC3) and non-prostate cell lines (NIH3T3 and HeLa). PCA3 expression was determined using the oligonucleotide primers described in the Methods section. PCA3 relative expression levels were determined in each cell line and compared to PCA3 expression in the DU145 cell line, used as a reference in this assay. (B) PCA3 expression was evaluated in LNCaP cells after knockdown using three different siPCA3 RNA sequences, termed siPCA3/1, siPCA3/2, and siPCA3/3. PCA3 expression was evaluated at 36 h post-transfection, and its relative expression level was determined compared to LNCaP cells transfected with the siSCr sequence, used as a negative control in this assay. (C) Following transfection of LNCaP cells with siPCA3/2, transcript levels were evaluated by qRT-PCR assays at the indicated time points. PCA3 relative expression is shown compared to siScr/LNCaP transfected cells. (D) PC3 cells were also transfected with siPCA3/2, and PCA3 expression was evaluated by qRT-PCR after 36 h post-transfection. 18S RNA was used as a constitutive gene. Data are shown as mean ± SD. All experiments were biological replicates, repeated a minimum of three times. **, p < 0.0017 and *** p < 0.0008.
Ferreira et al. BMC Cancer 2012 12:507 doi:10.1186/1471-2407-12-507