碩士 / 中國文化大學 / 化學系應用化學碩士班 / 104 / Zebrafish (Danio rerio) is one of the animal models for developmental biology that flourished in recent years. Compared to other vertebrates﹐zebrafish is characterized by short growth cycle, transparent embryos, easy breeding, and many kinds of mutation for genetic screening. It is widely used in embryonic development, drug screening, toxicology testing and identifying human-related diseases.
Every natural medicine has its pharmaceutical pathways; however, it may be harmful when treated in any condition. Before launching drugs for any purposes, repeated experiments and animal trials are necessary to examine if drugs have any side effects and harmfulness. In previous experiment, heral medicine 38, crude extract of one of native herbs in Taiwan, has been identified some anti-cancer effects on human lung cancer cells. It inhibited cell growth and increased cell apoptosis, Which indicates hm38 has potential anti-cancer activity.
We separated male and female fishes at night before setting up ezperiment. In the next morning, fishes were prompting and hour-post-fertilization (0 hpf) embryos were collected. Different concentrations of hm38: 0%, 0.5%, 2.5%, 10% were treated on embryos for 72 hours. In our preliminary, we observed that when increasing hm38 concentration, embryos have phenomenon of delayed development at 2.5% and 10%, and mortality increased dramatically; however, when treated embryos with 0.5% hm38, thereis no significant toxicity and growth retardation. Other non-specific variations may occur during treatment, such as the pericardium edema, which indicates hm38 for early zebrafish development has no significant toxicity. We subsequently test the signal molecules involving in anti-cancer pathway. We perform realtime-PCR analysis to screen the potential pathway that is affected by hm38. The mRNA expression levels of p53 is induced while BCL2 is significantly reduced, which suggests hm38 induces apoptosis through p53-mediated pathway by activating p53 and inhibiting BCL2 function. On other hand, we test the gene expression of SOX4a and 4b. While SOX4 is an oncogene, there is mounting evidence suggesting a role for SOX4 involvement in tumorigenes. Based on the mRNA expression pattern, SOX4a and 4b may exert potential anti-cancer progression function. In addition, protein level of caspase-3 and p53 are increased after 72 hrs treating 0.5% hm38, which correspond to our realtime-PCR results.
According to our results, this study demonstrates the feasibility of this novel combinational approach and shows that hm38 has potential to be anti-cancer agents.
| Identifer | oai:union.ndltd.org:TW/104PCCU0500015 |
| Date | January 2016 |
| Creators | Zheng-Hao Huang, 黃正豪 |
| Contributors | Yu-Heng Lai, 賴昱衡 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 35 |
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