PCA3 long noncoding RNA modulates the expression of key cancer-related genes in LNCaP prostate cancer cells
Prostate cancer antigen 3 (PCA3) is a prostate-specific long noncoding RNA (lncRNA) involved in the control of prostate cancer (PCa) cell survival, through modulating androgen receptor (AR) signaling. To further comprehend the mechanisms by which PCA3 modulates LNCaP cell survival, we characterized the expression patterns of several cancer-related genes, including those involved in epithelial-mesenchymal transition (EMT) and AR cofactors in response to PCA3 silencing. We also aimed to develop a strategy to stably silence PCA3. Small interfering RNA (siRNA) or short hairpin RNA (shRNA) was used to knock down PCA3 in LNCaP cells. The expression of 84 cancer-related genes, as well as those coding for AR cofactors and EMT markers, was analyzed by quantitative real-time PCR (qRT-PCR). LNCaP-PCA3 silenced cells differentially expressed 16 of the 84 cancer genes tested, mainly those involved in gene expression control and cell signaling. PCA3 knockdown also induced the upregulation of several transcripts coding for AR cofactors and modulated the expression of EMT markers. LNCaP cells transduced with lentivirus vectors carrying an shRNA sequence targeting PCA3 stably downregulated PCA3 expression, causing a significant drop (60 %) in the proportion of LNCaP cells expressing the transgene. In conclusion, our data provide evidence that PCA3 silencing modulates the expression of key cancer-related genes, including those coding for AR cofactors and EMT markers. Transducing LNCaP cells with an shRNA sequence targeting PCA3 led to loss of viability of the cells, supporting the proposal of PCA3 knockdown as a putative therapeutic approach to inhibit PCa growth.
KeywordsPCA3 Long noncoding RNA Gene expression Cancer-related genes Prostate cancer
This study was supported by grants from the Fundação de Amparo à Pesquisa do Rio de Janeiro (FAPERJ), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Instituto Nacional de Ciência e Tecnologia (INCT) para Controle do Câncer, Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Ministério da Saúde (MS)/Fundação do Câncer, Instituto Nacional de Câncer (INCa), Fundação Oswaldo Cruz (FIOCRUZ)/Bio-Manguinhos, and Swiss Bridge Foundation. We are especially grateful to Dr. Marcos Freire and Dr. Marco Medeiros for kindly supporting this study. We also thank Dr. Marcelo Ribeiro Alves for his kind support with statistical analyses.
Conflicts of interest
- 10.Sabbah M, Emami S, Redeuilh G, Julien S, Prévost G, Zimber A, et al. Molecular signature and therapeutic perspective of the epithelial-to-mesenchymal transitions in epithelial cancers. Drug Resist Update Rev Comment Antimicrob Anticancer Chemother. 2008;11:123–51.Google Scholar
- 11.Nantajit D, Lin D, Li JJ. The network of epithelial-mesenchymal transition: potential new targets for tumor resistance. J. Cancer Res Clin Oncol. 2014.Google Scholar
- 16.Paoli P, Giannoni E, Chiarugi P. Anoikis molecular pathways and its role in cancer progression. Biochim Biophys Acta. 1833;2013:3481–98.Google Scholar
- 31.Papadopoulou N, Charalampopoulos I, Anagnostopoulou V, Konstantinidis G, Föller M, Gravanis A, et al. Membrane androgen receptor activation triggers down-regulation of PI-3K/Akt/NF-kappaB activity and induces apoptotic responses via Bad, FasL and caspase-3 in DU145 prostate cancer cells. Mol Cancer. 2008;7:88.CrossRefPubMedPubMedCentralGoogle Scholar
- 32.Moehren U, Papaioannou M, Reeb CA, Grasselli A, Nanni S, Asim M, et al. Wild-type but not mutant androgen receptor inhibits expression of the hTERT telomerase subunit: a novel role of AR mutation for prostate cancer development. FASEB J Off Publ Fed Am Soc Exp Biol. 2008;22:1258–67.Google Scholar
- 38.Liu Z, Hao C, Song D, Zhang N, Bao H, Qu Q. androgen receptor coregulator CTBP1-as is associated with polycystic ovary syndrome in Chinese women: a preliminary study. Reprod Sci. Thousand Oaks Calif. 2014.Google Scholar