Experimental Study On Seversing Effect Of JSB-1 Conjugated ADM-loaded Immuno Nanoparticles On Multidrug Resistant Cell Line GRC-1 In Vitro

Objective: To prepare JSB-1 conjugated ADM-loaded immuno nanoparticle(JSB-1ADMNP), and examine the peculiar cytotoxicity of immuno nanoparticle ontargeting cells line GRC-1.To study on seversing effect of ADM-loaded immunonanoparticles with JSB-1 on multidrug resistant cells line GRC-1 in vitro.Methods:Determining the adriamycin(ADM) dosage of ADM-loadednanoparticle(ADMNP) by visible and ultraviolet spectroscopy. Making intoADM-loaded immuno nanoparticles by conjugating JSB-1 monoclonal antibodiesthrough glutaraldehyde, then the free JSB-1 monoclonal antibody molecules wasseparated from ADM-loaded immuno nanoparticles on a sephadex G-200minicolumn. The peculiar cytotoxicity of JSB-1ADMNP and ADMNP andreversion of MDR on GRC-1 cell line was studied by measuring the inhibition ofcell growth using the MTT assay, and the amount of intracellular ADMaccumulation was determined by flow cytometry.Results: The entrapping of ADM in ADM-loaded nanoparticle(ADMNP)is 0.78mg/ml. The nanoparticle were linked with JSB-1 monoclonalantibody(JSB-1ADMNP). Using GRC-1 cells as targeting cells and normalperipheral lymphocytes as control-group cells, we examined the peculiarcytotoxicity of JSB-1ADMNP on targeting cells and control-group cellsrespectively. The death rate of targeting cells after action for 96 hours is 49percent , whereas that of control-group cells is only 11 percent, from which weknow that the nanoparticle entrapping ADM bearing JSB-1 (JSB-1ADMNP) cankill their targeting cells peculiarly. The amount of intracellular ADM accumulationafter JSB-1ADMNP act on GRC-1 cells is 8.14 times than the ADM. Conclusions: JSB-1ADMNP are peculiar to their targeting cells and it canbeeffective to reverse multidrug resistance of the kidney cancer cells line GRC-1.JSB-1ADMNP showed an increased cytotoxicity towards GRC-1 cells comparedto ADMNP and ADM.