Development of Baculovirus Based-gene Therapy for Esophageal Cancer
| Autor: | YEN-JU CHAN, 詹燕茹 |
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| Rok vydání: | 2011 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 99 The baculovirus-mammalian gene expression vector system is a widely used and an important molecular biology tool. This is the first report on a novel esophageal cancer gene therapy based on photodynamic baculovirus-based model. Phosensitizers are small molecular photosensitive proteins used for photodynamic therapy（PDT）. In 2005, a light sensitive gene, KillerRed was discovered from Antomedusae chromoproteinam 2CP. The Killer Red protein has an excitation wavelength of around 540-580 nm that can produce reactive oxygen species（ROS）and induce cell death. In this study, the baculovirus-based bicistronic vectors（vCMV-KillerRed-EV71 IRES-EGFP, vCMV-EGFP -EV71 IRES-KillerRed ）were generated and were transduced to esophageal cancer cell lines （CE48T） to develop a vehicle for esophageal cancer gene therapy. Upon transduction, the photodynamic red fluorescent protein was irradiated by visible light to activate the KillerRed protein so as to induce cytotoxicity that is controlled by CMV promoter and EV71 IRES in vCMV-KillerRed-EV71 IRES-EGFP, vCMV-EGFP-EV71 IRES-KillerRed respectively. Furthermore, amantadine hydrochloride was used as an IRES regulatory compound to evaluate the activity of the CMV- EGFP-EV71 IRES-KillerRed on the esophageal cancer cell line. Hrer, if the amantadine is washed away, the system favors IRES dependent translation thus the cells appear worescent and the KillerRed gene will be translated resulting to toxicity on cells. Lastly, the study could be a promising tool for the development of a novel esophageal cancer gene therapy based on photodynamic model using a baculovirus expression system. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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