| Nowadays gene therapy is applied extensively in the field of tumors as well as disease of monogenic inheritance. Retinoblastoma, which is a typical malignant tumor caused by tumor suppressor gene inactivation, has become the hot spot of investigation in oapy, opthalmectomy, etc. With the development of molecular biology and techniques of gene engineering, gene therapy of RB has become the focus of tumor investigation. Comprehensive studies are carried on about suicide-gene therapy and exciting outcomes have been obtained, while evaluation of other schemes of gene therapy is still at initial stage. Establishment of an ideal animal model of RB that is easy to observe, more similar to clinical manifestations is vital for understand of tumor's biologphthalmology, oncology, medical genetics, etc. Presently at different developmental stages of RB, series of methods of therapy are carried on including cryotherapy, photocoagulation, thermotherapy, charged-particle radiotherapy, plaque brachytherapy, chemotherical behavior and evaluation of therapeutic effects of radiotherapy, chemotherapy and so on. Moreover, this novel model will play a significant role in gene therapy of RB.With the gradual development of study on tumor rational, more and more attention is focused on anti-angiogenesis strategy. VEGF and VEGFR are regarded as an important target for biological anti-tumor therapy. Interruption of combination of VEGF and its receptors is an effective approach to inhibit angiogenesis. At present, study on soluble VEGFR is still at the pre-clinical phase. Therefore, strategy of anti-angiogenesis aimed at soluble VEGFR by highly effective gene transfer techniques is becoming a big focal point for gene therapy. Gene transfer is an vital approach for the therapy of inherited diseases and tumors in the future. However, the available techniques of gene transfer display different kinds of deficiency. Compared with the commonly used vectors, plasmid has the advantages of convenient, economic, inferior antigenicity, applicable repeatedly, etc. Electroporation is the most effective non-viral technique for exogenous gene transfer, and it has been applied extensively due to its convenience and high efficiency. In our study, we attempt to establish thenovel orthotopic models of RB and malignant choroidal melanoma and explore the role of VEGF/VEGFR in the growth of tumors. Moreover, we investigate the effects and feasibility of gene therapy of RB aiming at VEGF/VEGFR pathway by electroporation.Methods1. Establishment of a novel orthotopic retinoblastoma model expressing green fluorescent protein. 2l HXO-Rb44-GFP cell suspension (density as 4.5 X 108/ml~ 5.5 108/ml ) were injected into the subretinal cavity of 30 nude mice (60 eyes) under binocular operating microscope. The growth of the transplanted RB was observed in vivo under the fluorescence stereomicroscope. The nude mice were killed at different time post-operation to investigate the metastasis of the tumor to optic nerve, brain , and other organs including lung, liver, spleen and kidney. Moreover, pathological detection and immunohistochemical staining of GFP were carried out.2. Establishment of a novel orthotopic model of malignant choroidal melanoma expressing green fluorescent protein. 21 OCM-1-GFP cell suspension (density as 4.5 107/ml~5.5 107/ml) was injected into the subretinal cavity of right eye of 40 nude mice (40 eyes) under binocular operating microscope. The growth of the transplanted malignant choroidal melanoma was observed in vivo under the fluorescence stereomicroscope. The nude mice were killed at intraocular phase, extraocular phase, and failure phase respectively to detect the metastasis of the tumor to optic nerve, brain and other organs including lung, liver, kidney and spleen. Moreover, pathological detection and immunohistochemical staining of GFP was carried out.3. Study on the difference of biological behavior between transplanted RB and malignant choroidal melanoma. Tumor cell lines including HXO-Rb44-GFP HXO-Rb44 OC...
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