The Construction Of A Novel C-PC/CMC-CD59sp Nanoparticles And Its Targeted Antitumor Study For HeLa Cells

Objective:Carbocymethyl chitosan(CMC) loading C-phycocyanin(C-PC) with the lead of CD59 specific ligand peptide(CD59sp) nanoparticles(C-PC/CMC-CD59sp-NPs) were made. The characteristics and mechanism of targeted anti-tumor were explored in order to realize the target therapy of C-PC in He La cells.Methods:The targeting nanoparticles were synthesized by ionic-gelation method, and the optimal condition was selected out by orthogonal analysis. The properties of nanoparticles were observed by transmission electron microscopy(TEM) and dynamic light scattering(DLS) and Fourier Transform Infrared Spectrometer(FTIR). The effects of nanoparticles on the He La cells proliferation were assessed by MTT assay. The safety was studied by hemolysis assay and targeted uptake was observed using by the confocal microscopy. The expression quantities of cleaved caspase-3, cleaved PARP, Bcl-2 were determined by western blot and immunofluorescent staining. The cell cycle was detected by Flow Cytometry(FCM).Results : The preparing conditions of nanoparticles were optimized to make ideal C-PC/CMC-CD59sp-NPs. The optimal conditions were as follows : the quantity of CMC/C-PC =3:1, the concentration of CMC=2.0 mg/ml and Ca Cl2=1.0 mg/ml. The average size of the nanoparticles was about 200 nm with spherical morphology. The entrapment efficiency was about 65% and drug-loading capacity was 20%. The drug-loading nanoparticles showed some characteristics such as slow release in vitro and good hemocompatibility. Under the guide of CD59 sp, the targeting nanoparticles could arrive on surface of He La cells targetedly and efficiently, and they could obviously inhibit He La cells proliferation. The nanoparticles also induced more apoptosis by up-regulating cleaved caspase-3 and cleaved PARP proteins, and down-regulating of Bcl-2 proteins.The cell cycle was significantly arrested, mostly at G1 phase.Conclusion:The targeted C-PC/CMC-CD59 sp nanoparticles were successfully prepared,possessing the properties of targeting, slow release and safety. The antitumor effects of nanoparticles were realized by inhibiting He La cells proliferation, inducing cells apoptosis, and arresting cell cycle process at G1 phase. Our study provided a new idea for the research and development of marine drugs, so did a theoretical support for the targeted therapy of anticancer drug.