title: Knockdown of PSCA induces EMT and decreases metastatic potentials of the human prostate cancer DU145 cells.
abstract: BACKGROUND
  Prostate stem cell antigen (PSCA) expression has been shown to correlate with prostatic carcinogenesis and prostate cancer (PCa) progression. The underlying mechanisms for these processes are currently unknown. Epithelial to mesenchymal transition (EMT) has been associated with the invasiveness and the distant metastasis of PCa. In this study, we investigated the effects of knocking down the PSCA on the cell migration, the invasiveness, and the EMT of the PCa cell line DU145 in vitro and in vivo.
  
  
  METHODS
  Four target sequences of the small hairpin RNA for PSCA were designed, and the best effect knockdown sequence shRNA#1 was screened to construct the stable transfected DU145 cell line (DU145 shRNA#1), the scramble sequence was also designed to construct the stable transfected DU145 cell line(DU145 scramble). Cell migration and invasion were studied using Transwell assay. Quantitative RT-PCR, Western blot (WB) were used to quantify PSCA, E-cadherin, β-catenin, Vimentin, Fibronectin expression in DU145, DU145 scramble, DU145 shRNA#1 in vitro and in vivo. RT-PCR, immunofluorescent staining were used to quantify PSCA, E-cadherin, and Vimentin expression in vitro. EMT-related genes Snail, Slug, and Twist, were quantified by quantitative RT-PCR in vitro.
  
  
  RESULTS
  The constructed stable knockdown of the PSCA in the DU145 cell had a silencing effect up to 90.5 %. DU145 shRNA#1 became scattered from the tightly packed colonies. It was associated with decreased cell migration and invasion. There was also an increased Vimentin and Fibronectin expression, an inhibited E-cadherin and β-catenin expression at both the mRNA and the protein levels when compared to the DU145 and the DU145 scramble in vitro and vivo. Furthermore, with the exception of the Snail, the expression of EMT-related Slug and Twist genes were upregulated.
  
  
  CONCLUSIONS
  Our data indicated that knockdown of PSCA induced EMT and reduced metastatic potentials of the DU145 cells, suggesting that PSCA played an important role in prostatic carcinogenesis and progression.
authors.affiliation: Department of Urology & Andrology, Minimally Invasive Surgery Center, The First Affiliated Hospital of Guangzhou Medical University, Guangdong Provincial Key Laboratory of Urology, No.1-3, Kangda Road, Guangzhou, 510230 Guangdong Province China.
authors.forename: Ran
authors.initials: R
authors.lastname: Kang
authors.affiliation: Department of Urology & Andrology, Minimally Invasive Surgery Center, The First Affiliated Hospital of Guangzhou Medical University, Guangdong Provincial Key Laboratory of Urology, No.1-3, Kangda Road, Guangzhou, 510230 Guangdong Province China.
authors.forename: Shankun
authors.initials: S
authors.lastname: Zhao
authors.affiliation: Department of Urology & Andrology, Minimally Invasive Surgery Center, The First Affiliated Hospital of Guangzhou Medical University, Guangdong Provincial Key Laboratory of Urology, No.1-3, Kangda Road, Guangzhou, 510230 Guangdong Province China.
authors.forename: Luhao
authors.initials: L
authors.lastname: Liu
authors.affiliation: Department of Urology & Andrology, Minimally Invasive Surgery Center, The First Affiliated Hospital of Guangzhou Medical University, Guangdong Provincial Key Laboratory of Urology, No.1-3, Kangda Road, Guangzhou, 510230 Guangdong Province China.
authors.forename: Futian
authors.initials: F
authors.lastname: Li
authors.affiliation: Department of Urology & Andrology, Minimally Invasive Surgery Center, The First Affiliated Hospital of Guangzhou Medical University, Guangdong Provincial Key Laboratory of Urology, No.1-3, Kangda Road, Guangzhou, 510230 Guangdong Province China.
authors.forename: Ermao
authors.initials: E
authors.lastname: Li
authors.affiliation: Department of Urology & Andrology, Minimally Invasive Surgery Center, The First Affiliated Hospital of Guangzhou Medical University, Guangdong Provincial Key Laboratory of Urology, No.1-3, Kangda Road, Guangzhou, 510230 Guangdong Province China.
authors.forename: Lianmin
authors.initials: L
authors.lastname: Luo
authors.affiliation: Department of Hematology, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510230 China.
authors.forename: Lihua
authors.initials: L
authors.lastname: Xu
authors.affiliation: Department of Urology & Andrology, Minimally Invasive Surgery Center, The First Affiliated Hospital of Guangzhou Medical University, Guangdong Provincial Key Laboratory of Urology, No.1-3, Kangda Road, Guangzhou, 510230 Guangdong Province China.
authors.forename: ShawPong
authors.initials: S
authors.lastname: Wan
authors.affiliation: Department of Urology & Andrology, Minimally Invasive Surgery Center, The First Affiliated Hospital of Guangzhou Medical University, Guangdong Provincial Key Laboratory of Urology, No.1-3, Kangda Road, Guangzhou, 510230 Guangdong Province China.
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Zhao
keywords: Epithelial-mesenchymal transition; Metastasis; Prostate cancer; Prostate stem cell antigen