| Summary: | 碩士 === 國立臺南大學 === 生物科技學系碩士班 === 101 === Cancer has lasted for 31 years on the top ten of domestic leading death. There are several strategies for cancer treatment, including surgical resection, chemotherapy, and radiotherapy. Chemotherapeutic agents not only have various side effects, but also have considerable burden in patients. Multidrug resistance (MDR) is one of the major obstacles to successful chemotherapy. It is often related to overexpression of drug efflux pumps, such as P-glycoprotein (P-gp) that dramatically reduces intracellular drug concentrations and causes pump resistance. Concurrently, the development of antiapoptotic survival pathways in cancer cells may cause non-pump MDR. Therefore, we focus on exploring the new cancer treatment regimen. In the present research, we use epirubicin (Epi) as a model drug to combine with one antimicrobial peptide, P. In the MTT assay, we found that, P can effectively reduce the survival rate of cancer cells, combined Epi treatment, the survival rate of cancer cells more effectively reduced. In the caspase assay, we found that, P can effectively improve the caspase 3, caspase 8 and caspase 9 activity, Epi can increase caspase 3 and caspase 9 activity, but does not affect the activity of caspase 8; flow cytometric analysis of intracellular accumulation P and the combination experiments after Epi can effectively improve the intracellular accumulation amount; the cell cycle analysis, combined with P and the Epi treatment, sub-G1 phase of the cell will increase substantially. From the above results, we can see that, P, and Epi can effectively induce apoptosis in cells and, therefore, we use the qPCR to observe the P Epi cells for three different change in the amount of mRNA expression.
Moreover, according to literature reports, zebrafish can be developed as a new drug screening model. In the present study, we examine the influence of Epi on zebrafish using morphology observation and heart toxicity test. Finally, we use qPCR to analyze the effect of Epi on MDR transporter-related and apoptosis-related genes of zebrafish. In the morphology observation test, we found the Epi not only make the zebrafish shrink to lessen, even in the high concentration also cause deformities tail. In addition, it will result in the shrinking of the zebrafish heart. Therefore, development of a new drug to clinical needs a rather long time, but can be directly through the understanding of the drug zebrafish of the organism, not only to compensate for the lack of cell experiments can also get information vivo. With the rapid high-throughput screening, to obtain embryo survival as measured by MTT assay Cell viability in drug can simultaneously influence the living body, and can also be mRNA level of testing, even in a short time can be observed Drug-induced apoptosis and activation of p53 protein. Thus, we use zebrafish as an in vivo platform to get more information about anticancer therapy. We expect that the drug screening model of zebrafish can provide an easier, faster, and more accurate detection of chemotherapy treatment regimen.
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