Abstract
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.
Highlights
Prostate cancer is the most common cancer among men worldwide
As there is no current mouse model that fully recapitulates all the features of prostate cancer [2], we aimed to establish a transgenic mouse model of prostate cancer using the Sleeping Beauty we aimed to establish a transgenic mouse model of prostate cancer using the Sleeping Beauty transposon system enhanced by electroporation in this work
Transposon system enhanced by electroporation in this work
Summary
Prostate cancer is the most common cancer among men worldwide. There are currently no mouse models that fully recapitulate the features of human prostate cancer, as spontaneously occurring prostate cancer is uncommon in mice [1,2]. In a 2-year toxicology and carcinogenicity study conducted by the National Toxicology Program, 612 control B6C3F1 mice failed to develop neoplastic lesions [3]. There is a great need for a mouse model of prostate cancer. We used the Sleeping Beauty (SB) transposon system combined with electroporation to establish a mouse model of prostate cancer. The SB transposon, which was genetically engineered for the purpose of insertional mutagenesis [4], is a member of the Tc1/mariner family [5]. Transposons are DNA pieces that are flanked by terminal
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