RESUMEN
OBJECTIVE: To generate a sensitive tool for noninvasive monitoring of a therapeutic gene vasostatin. METHODS: We fused the bioluminescent reporter gene firefly luciferase to the therapeutic transgene vasostatin and ensured that these two proteins would not interrupt each other and kept their own natural character. RESULTS: We therefore examined clones of PC3 cells stably expressing fusion gene and positive controlfluc with bioluminescence. In vivo imaging of PC3-Fluc subcutaneous tumors showed that the mean tumor bioluminescence increased in animals over several weeks. CONCLUSION: Noninvasive monitoring facilitates the detection of gene expression in vivo and in vitro.
Asunto(s)
Calreticulina/metabolismo , Fragmentos de Péptidos/metabolismo , Animales , Calreticulina/genética , Línea Celular Tumoral , Técnicas de Transferencia de Gen , Genes Reporteros , Humanos , Luciferasas de Luciérnaga/genética , Luciferasas de Luciérnaga/metabolismo , Mediciones Luminiscentes , Trasplante de Neoplasias , Fragmentos de Péptidos/genética , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismoRESUMEN
OBJECTIVE: The synthesis, biodistribution, and animal imaging of 99mTc- hydrazinonicotinamide-folate (99mTc-HYNIC-Folate) were studied as a folate receptor-targeted tumor imaging agent. METHODS: HYNIC-Folate was synthesized by a muti-step reaction and radiolabeled with 99mTc using tricine and trisodium phenylphosphine-3, 3', 3"-trisulfonate (TPPTS) as coligands. The radiochemical purity and stability of 99mTc HYNIC-Folate was measured. The biodistributions of 99mTc-HYNIC-Folate in normal mice and tumor-bearing mice were detected. Whole-body gamma imaging was performed using an athymic mouse tumor xenograft model. RESULTS: The ligand HYNIC-Folate was successfully synthesized and characterized by hydrogen nuclear magnetic resonance (1HNMR) and mass spectrometry (MS). The radiochemical purity of 99mTc-HYNIC-Folate was 96% under optimal conditions. Data from gamma scintigraphy and the biodistribution in tumor-bearing mice showed that 99mTc-HYNIC-Folate predominantly accumulated in tumor, its uptake rate per gram tissue alpham was 5. 620+/- 0. 753. The uptakes of 99mTc-HYNIC-Folate in the other non-target tissues were very low, except it was high in the kidneys ( am was 41. 959 +/-6. 759) . CONCLUSION: 99mTc-HYNIC-Folate has the potential to be used as a noninvasive radiodiagnostic imaging agent for the detection of folate receptor-positive human cancers.
Asunto(s)
Compuestos de Organotecnecio/síntesis química , Radiofármacos/síntesis química , Animales , Femenino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos ICR , Neoplasias Experimentales/diagnóstico por imagen , Compuestos de Organotecnecio/farmacocinética , Cintigrafía , Radiofármacos/farmacocinética , Distribución TisularRESUMEN
OBJECTIVE: To express and purify the recombinant human pigment epithelium-derived factor (PEDF) which inhibits the proliferation of the endothelium cells from blood vessel in E.coli. METHODS: PEDF gene was inserted into the prokaryotic expression vector pGEX-4T-2. The recombinant protein PEDF was expressed in E.coli BL-21, and purified by the GST Sepharose 4B affinity column. The recombinant human PEDF protein was identified by Western blot and mass spectrum. The biological activity of the recombinant human PEDF protein was measured by using MTT. RESULTS: The 46 kDa recombinant human PEDF protein was obtained. It significantly inhibited the proliferation of the human umbilical vein cell line HUVEC. CONCLUSION: The recombinant human PEDF with anti-angiogenesis activity protein may be successfully purify through prokaryotic expression.