| Summary: | 碩士 === 長庚大學 === 生物醫學研究所 === 100 === Cancer is one of the most common causes of death, which can be induced by dysregulation of oncogenes, tumor suppressor genes and/or microRNAs. To evaluate the oncogenic transformation potential of putative oncogenes, methods including transgenic, gene targeting, and xenograft approaches in rodent have been well-established for years. However, the rodent model faces the constraints on high cost, intensive human resources and larger spaces, and longer time to evaluate the biological effects. Hence, a small animal model that can quickly assay the effects of putative oncogenes with lower cost is considered as a good alternative one. Using a gain-of-function approach to overexpress oncogenes in the keratinocytes of zebrafish, the morphological transformation in the keratinocytes can be non-invasively observed. In this thesis, the human oncogenes were driven by a skin-specific promoter, and used the gateway recombination and Tol2 transposon system to assemble the expression cassettes. In this high-throughput platform, the expressional vectors performed in transient or stable assay in zebrafish skin. Consequently, the zebrafish skin is quite refractory to single exogenesis of oncogenes. To induce the transformation of skin cancer with success, oncogenes were co-activated in a transcription factor-overexpressing transgenic line (driver line). The results successfully identified the oncogenic transformation at 7 days post-fertilization when double oncogenic pathways are co-activated in zebrafish skin. The oncogenic driver line established in this study provides an alternative and quick approach to test potential oncogene in living animal.
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