RESUMO
Multi-drug resistance (MDR) is a phenomenon that tumor cells are exposed to a chemotherapeutic drug for a long time and then develop resistance to a variety of other anticancer drugs with different structures and different mechanisms. The in vitro studies of tumor cell lines cannot systematically reflect the role of MDR gene in vivo, and the cost of in vivo studies of transgenic mice as animal models is high. Given the myriad merits of zebrafish relative to other animal models, we aimed to establish a screening system using zebrafish stably expressing ATP-binding cassette (ATP-cassette) superfamily transporters and unveil the potential regulatory mechanism. We first used the Tol2-mediated approach to construct a Tg (abcb4:EGFP) transgenic zebrafish line with ATP-binding cassette (ABC) subfamily B member 4 (abcb4) gene promoter to drive EGFP expression. The expression levels of abcb4 and EGFP were significantly increased when Tg(abcb4:EGFP) transgenic zebrafish embryos were exposed to doxorubicin (DOX) or vincristine (VCR), and the increases were accompanied by a marked decreased accumulation of rhodamine B (RhB) in embryos, indicating a remarkable increase in DOX or VCR efflux. Mechanistically, Akt and Erk signalings were activated upon the treatment with DOX or VCR. With the application of Akt and Erk inhibitors, drug resistance was reversed with differing responsive effects. Notably, downstream NF-κB played a central role in the regulation of abcb4-mediated drug resistance. Taken together, the data indicate that the engineered Tg(abcb4:EGFP) transgenic zebrafish model is a new platform for screening drug resistance in vivo, which may facilitate and accelerate the process of drug development.
Assuntos
Transportadores de Cassetes de Ligação de ATP , NF-kappa B , Proteínas de Peixe-Zebra , Peixe-Zebra , Transportadores de Cassetes de Ligação de ATP/genética , Transportadores de Cassetes de Ligação de ATP/metabolismo , Animais , Animais Geneticamente Modificados , Linhagem Celular Tumoral , Doxorrubicina/farmacologia , Resistência a Medicamentos , Resistencia a Medicamentos Antineoplásicos , NF-kappa B/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Vincristina/farmacologia , Peixe-Zebra/genética , Peixe-Zebra/metabolismo , Proteínas de Peixe-Zebra/genética , Proteínas de Peixe-Zebra/metabolismoRESUMO
BACKGROUND: Based on the background of minimally invasive surgery and applications of medical robots, a vascular interventional robotic system has been developed that can be used in the field of vascular intervention. METHODS: The robotic system comprises a propulsion system, an image navigation system and a virtual surgery training system. Integration of the three systems constitutes a vascular intervention prototype robotic system used to carry out in vitro vascular intervention and animal experiments. RESULTS: On a transparent glass vascular model, a catheter was shown to enter an arbitrary branch of the vascular model with catheter motion meeting the requirements of clinical vascular intervention surgery (VIS); i.e. error band of catheter motion < 0.5 mm. In the animal experiments, 1.33-2.00 mm (4F-6F) diameter catheters were selectively inserted successfully into predefined targets in the animal, such as the renal, cardiovascular and cerebrovascular artery. Compared with conventional manual surgery, the time for robotic surgery is a little longer. There were no operative complications in the animal experiments. CONCLUSIONS: These simulation and animal study results demonstrate that this vascular interventional robotic system allows doctors to perform angiography remotely and prevents them from radiation exposure. The system may be the basis for further clinical applications of vascular intervention.