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Knockdown of PSCA induces EMT and decreases metastatic potentials of the human prostate cancer DU145 cells
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 invas...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4791869/ https://www.ncbi.nlm.nih.gov/pubmed/26981049 http://dx.doi.org/10.1186/s12935-016-0295-4 |
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author | Kang, Ran Zhao, Shankun Liu, Luhao Li, Futian Li, Ermao Luo, Lianmin Xu, Lihua Wan, ShawPong Zhao, Zhigang |
author_facet | Kang, Ran Zhao, Shankun Liu, Luhao Li, Futian Li, Ermao Luo, Lianmin Xu, Lihua Wan, ShawPong Zhao, Zhigang |
author_sort | Kang, Ran |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-4791869 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-47918692016-03-16 Knockdown of PSCA induces EMT and decreases metastatic potentials of the human prostate cancer DU145 cells Kang, Ran Zhao, Shankun Liu, Luhao Li, Futian Li, Ermao Luo, Lianmin Xu, Lihua Wan, ShawPong Zhao, Zhigang Cancer Cell Int Primary Research 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. BioMed Central 2016-03-15 /pmc/articles/PMC4791869/ /pubmed/26981049 http://dx.doi.org/10.1186/s12935-016-0295-4 Text en © Kang et al. 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Primary Research Kang, Ran Zhao, Shankun Liu, Luhao Li, Futian Li, Ermao Luo, Lianmin Xu, Lihua Wan, ShawPong Zhao, Zhigang Knockdown of PSCA induces EMT and decreases metastatic potentials of the human prostate cancer DU145 cells |
title | Knockdown of PSCA induces EMT and decreases metastatic potentials of the human prostate cancer DU145 cells |
title_full | Knockdown of PSCA induces EMT and decreases metastatic potentials of the human prostate cancer DU145 cells |
title_fullStr | Knockdown of PSCA induces EMT and decreases metastatic potentials of the human prostate cancer DU145 cells |
title_full_unstemmed | Knockdown of PSCA induces EMT and decreases metastatic potentials of the human prostate cancer DU145 cells |
title_short | Knockdown of PSCA induces EMT and decreases metastatic potentials of the human prostate cancer DU145 cells |
title_sort | knockdown of psca induces emt and decreases metastatic potentials of the human prostate cancer du145 cells |
topic | Primary Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4791869/ https://www.ncbi.nlm.nih.gov/pubmed/26981049 http://dx.doi.org/10.1186/s12935-016-0295-4 |
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