Inhibition Of 4T₁-Luc On Breast Cancer In Vitro Based On ABD-iTEP-PTX Nanoparticles

Objective: Immunogen-free elastin(ABD-ITEP)nanocarriers were constructed and self-assembled with modified paclitaxel(PTX-LEV-EMCH)to form nanoparticles.The effects of drugs on tumor cell proliferation inhibition were compared and evaluated in vitro.Methods: ABD-ITEP nanocarrier was prepared and identified according to the characteristics of elastin polypeptide,and then self-assembled with modified paclitaxel to form nano drug particles(ABD-ITEP PTX NPS)by chemical bond mercapto group link.Protein-binding nanomedics(HSA-ABD-ITEP-PTX NPS)were formed by the easy binding of protein domain(ABD)to human serum albumin(HSA).The particle size and Zeta potential of the constructed nano-drugs were tested by Nanobrook Omni.TEM images were obtained under a transmission electron microscope,and Nano Measurer randomly counted the particle size distribution of the nanoparticles in the TEM images.The encapsulation rate and drug loading rate were calculated by ultraviolet spectrophotometer.The cytotoxicity of HSA-ABD-ITEP-PTX NPS and ABD-ITEP was evaluated by CCK-8 cell activity assay.The in vitro release of drug-loaded nanoparticles was detected by simulating the difference between normal tissue environment and acidic tumor tissue environment.Results: ABD-ITEP-PTX NPS and HSA-ABD-ITEP-PTX NPS TEM images showed the typical spherical elliptical morphology of nanoparticle particles.The average particle size of ABD-ITEP-PTX NPS was 86.75 nm(PDI: 0.16)and the Zeta potential showed no charge;The average particle size of HSA-ABD-ITEP-PTX NPS was 115.76 nm(PDI: 0.17),and the Zeta potential showed a negative charge of-22.54±1.06 m V.The encapsulation rate and drug loading rate were 51.40 % and18.78 %,respectively.No cytotoxicity was observed in the prepared ABD-ITEP nanocarrier at 62.5 ～ 500 μg/m L.HSA-ABD-ITEP-PTX NPS significantly inhibited breast tumor 4T1-Luc cells,significantly higher than the control group of free paclitaxel.The IC50 was 9.6 μg/m L,which was higher than that of albumin-bound paclitaxel.ABD-ITEP-PTX NPS has a high release rate in relatively normal tissues in more acidic environments.Conclusion: A novel self-assembled paclitaxel nano drug(HSA-ABD-ITEP-PTX NPS)was successfully constructed using immunogen-free elastin polypeptide as carrier,and it effectively inhibited tumor cell proliferation in vitro.This study laid a foundation for the further study of HSA-ABD-ITEP-PTX NPS in small animal experiments.