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Development of a Mouse Model of Prostate Cancer Using the Sleeping Beauty Transposon and Electroporation

The Sleeping Beauty (SB) transposon system is non-viral and uses insertional mutagenesis, resulting in the permanent expression of transferred genes. Although the SB transposon is a useful method for establishing a mouse tumor model, there has been difficulty in using this method to generate tumors...

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Autores principales: Choi, Hyun-Ji, Lee, Han-Byul, Jung, Sunyoung, Park, Hyun-Kyu, Jo, Woori, Cho, Sung-Min, Kim, Woo-Jin, Son, Woo-Chan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6100630/
https://www.ncbi.nlm.nih.gov/pubmed/29874846
http://dx.doi.org/10.3390/molecules23061360
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author Choi, Hyun-Ji
Lee, Han-Byul
Jung, Sunyoung
Park, Hyun-Kyu
Jo, Woori
Cho, Sung-Min
Kim, Woo-Jin
Son, Woo-Chan
author_facet Choi, Hyun-Ji
Lee, Han-Byul
Jung, Sunyoung
Park, Hyun-Kyu
Jo, Woori
Cho, Sung-Min
Kim, Woo-Jin
Son, Woo-Chan
author_sort Choi, Hyun-Ji
collection PubMed
description The Sleeping Beauty (SB) transposon system is non-viral and uses insertional mutagenesis, resulting in the permanent expression of transferred genes. Although the SB transposon is a useful method for establishing a mouse tumor model, there has been difficulty in using this method to generate tumors in the prostate. In the present study, electroporation was used to enhance the transfection efficiency of the SB transposon. To generate tumors, three constructs (a c-Myc expression cassette, a HRAS (HRas proto-oncogene, GTPase) expression cassette and a shRNA against p53) contained within the SB transposon plasmids were directly injected into the prostate. Electroporation was conducted on the injection site after the injection of the DNA plasmid. Following the tumorigenesis, the tumors were monitored by animal PET imaging and identified by gross observation. After this, the tumors were characterized by using histological and immunohistochemical techniques. The expression of the targeted genes was analyzed by Real-Time qRT-PCR. All mice subjected to the injection were found to have prostate tumors, which was supported by PSA immunohistochemistry. To our knowledge, this is the first demonstration of tumor induction in the mouse prostate using the electroporation-enhanced SB transposon system in combination with c-Myc, HRAS and p53. This model serves as a valuable resource for the future development of SB-induced mouse models of cancer.
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spelling pubmed-61006302018-11-13 Development of a Mouse Model of Prostate Cancer Using the Sleeping Beauty Transposon and Electroporation Choi, Hyun-Ji Lee, Han-Byul Jung, Sunyoung Park, Hyun-Kyu Jo, Woori Cho, Sung-Min Kim, Woo-Jin Son, Woo-Chan Molecules Article The Sleeping Beauty (SB) transposon system is non-viral and uses insertional mutagenesis, resulting in the permanent expression of transferred genes. Although the SB transposon is a useful method for establishing a mouse tumor model, there has been difficulty in using this method to generate tumors in the prostate. In the present study, electroporation was used to enhance the transfection efficiency of the SB transposon. To generate tumors, three constructs (a c-Myc expression cassette, a HRAS (HRas proto-oncogene, GTPase) expression cassette and a shRNA against p53) contained within the SB transposon plasmids were directly injected into the prostate. Electroporation was conducted on the injection site after the injection of the DNA plasmid. Following the tumorigenesis, the tumors were monitored by animal PET imaging and identified by gross observation. After this, the tumors were characterized by using histological and immunohistochemical techniques. The expression of the targeted genes was analyzed by Real-Time qRT-PCR. All mice subjected to the injection were found to have prostate tumors, which was supported by PSA immunohistochemistry. To our knowledge, this is the first demonstration of tumor induction in the mouse prostate using the electroporation-enhanced SB transposon system in combination with c-Myc, HRAS and p53. This model serves as a valuable resource for the future development of SB-induced mouse models of cancer. MDPI 2018-06-05 /pmc/articles/PMC6100630/ /pubmed/29874846 http://dx.doi.org/10.3390/molecules23061360 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Choi, Hyun-Ji
Lee, Han-Byul
Jung, Sunyoung
Park, Hyun-Kyu
Jo, Woori
Cho, Sung-Min
Kim, Woo-Jin
Son, Woo-Chan
Development of a Mouse Model of Prostate Cancer Using the Sleeping Beauty Transposon and Electroporation
title Development of a Mouse Model of Prostate Cancer Using the Sleeping Beauty Transposon and Electroporation
title_full Development of a Mouse Model of Prostate Cancer Using the Sleeping Beauty Transposon and Electroporation
title_fullStr Development of a Mouse Model of Prostate Cancer Using the Sleeping Beauty Transposon and Electroporation
title_full_unstemmed Development of a Mouse Model of Prostate Cancer Using the Sleeping Beauty Transposon and Electroporation
title_short Development of a Mouse Model of Prostate Cancer Using the Sleeping Beauty Transposon and Electroporation
title_sort development of a mouse model of prostate cancer using the sleeping beauty transposon and electroporation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6100630/
https://www.ncbi.nlm.nih.gov/pubmed/29874846
http://dx.doi.org/10.3390/molecules23061360
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