Inhibitory Effect Of New ABD-iTEP-（G₆C）₈-Sal Nanoparticles On Breast Cancer Stem Cells In Vitro

Objective:To construct a novel non-immunogenic elastic polypeptide ABD-iTEP-（G₆C）₈nanoparticle system and carry the anti-tumor stem cell drug salinomycin to inhibit breast cancer in vitro with albumin paclitaxel.Methods:Firstly,a method for the determination of halomycin was established by ultraviolet spectrophotometry.Secondly,using artificial synthesis technology to design and synthesize c DNA transfection,transcription and translation to obtain albumin binding non-immunogenic elastic polypeptides:ABD-iTEP-（G₆C）₈,the hydrophilic and non-immunogenicity elastic peptide ABD-iTEP-（G₆C）₈nanoparticles were purified by polypeptide phase transition technique,and then the hydrophobic MPBH-Sal was bonded by hyzone bond coupling to form nanoparticles by self-assembly,and the nanoparticles ABD-iTEP-（G₆C）₈-Sal was obtained.And to optimize the production.The particle size and Zeta potential were measured by Nanobrook Omni,and the morphology of nanoparticles was obtained by transmission electron microscopy.The release regularity of ABD-iTEP-（G₆C）₈-Sal was obtained by in vitro release experiments.The affinity between HSA and ABD-iTEP-（G₆C）₈-Sal was determined by polyacrylamide gel electrophoresis.The in vitro antitumor activity of ABD-iTEP-（G₆C）₈-Sal and salinomycin were studied by cell proliferation and pellet formation experiments.Results:The standard curve for the determination of salinomycin was Y=0.07212×X+0.01093,and the concentration of MPBH-Sal showed a good correlation in the range of（0-20）μg/ml（R²=0.9996）.The nano-drug ABD-iTEP-（G₆C）₈-Sal has a particle size of about 89 nm and a Zeta potential of-10m V.By simulating tumor and normal tissue environment and using in vitro release experiment,it is concluded that the release rate of the compound can reach more than 80%in acid condition and less than 20%in neutral environment.According to polyacrylamide gel electrophoresis,HSA and ABD-iTEP-（G₆C）₈-Sal alwere fully combined when the molar ratio was 1:2.The anti-tumor activity invitro showed that ABD-iTEP-（G₆C）₈-Sal had stronger toxic effect on breast cancer microsphere cells and could obviously inhibit the growth of breast cancermicrosphere cells.Combined with Nab-PTX,it is proved that the combination of the two drugs has higher anti-tumor effect than the single use of the two drugs.Conclusion:The nanoparticle ABD-iTEP-（G₆C）₈-Sal was successfully synthesized,and the 4T1-Luc breast cancer cell model was established.The results showed that ABD-iTEP-（G₆C）₈-Sal had better targeted killing ability and higher biological safety.Combined with Nab-PTX,the killing effect on breast cancer cells was further enhanced.