| Summary: | 碩士 === 淡江大學 === 生命科學研究所碩士班 === 98 === According to previous studies that shh signaling pathway will continually be activated in many tumor cells and then cause cell carcinoma to lead into cancer. One of the basal cell skin cancer is basal cell carcinoma (BCC) which is caused by shh signaling pathway. In our previous studying, we found that when zebrafish transgenic lines Tg (K18: shh-RFP) Shh overexpress Shh in vivo, it will lead skin to have cancer phenomenon and then take the embryos for gene chip (microarray) analysis. We found rrm2 gene expression significantly increase in fish (Chen et al., 2009), so when shh is overexpressed , it will lead rrm2 into over-activation.
We generated the transgenic line Tg(K18:rrm2-RFP) that over-express rrm2 restrictively in the epidermis. We found some variation of the transgenic line by 4 days-post-fertilization(dpf), including that the mutation of head, eyes and the base skin of pectoral fin. Comparing with wild strain found that this phenotype is similar to the transgenic line Tg(K18:shh-RFP), then presume that may have the correlation between shh and rrm2.Through the experiments to test and verify the variety skin tissues that had different between Tg(K18:rrm2-RFP) and wild type(WT).To confirm the expression pattern of rrm2, we can use whole-mount in situ hybridization to display. Observing the expression pattern of rrm2,we found that rrm2 was expressed at the region of developing cell cycle and cell proliferation.
Futher,we designed a series of experiment to study the relevance of rrm2 and shh.We use cyclopamine and injected K18:shh:RFP plasmid to change shh expression in vivo.Silimarly,through observed the expression pattern of rrm2 demonstrate that shh was inhibited or overexpressed lead to rrm2 expressed addition or reduction.
If rrm2 was effected on shh,we tried to search the Gli binding site at rrm2 upstream or downstream sequences.Then,we found the silimar Gli binding site at rrm2 upstream sequences presumed that rrm2 would be regulated by shh.Shh was overexpressed that it increased rrm2 expression and caused skin carcinogenesis by inducing epidermal cells over-proliferation.In conclusion,we might discuss the connection between rrm2 and shh in this thesis.
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