| Objective : C-PC/CMC-CD59 sp nanoparticles have been successfully constructed in which CMC served as a carrier,C-PC acted as an anti-tumor drug,CD59 sp functioned as a targeted molecule.C-PC was delivered to the surface of cervical cancer cells via the targeted drug delivery system.Furthermore,we would like to prove that the anti-tumor effect and mechanism of C-PC/CMC and C-PC/CMC-CD59 sp NPs on cervical cancer(HeLa and Si Ha)in vitro and in vivo.Methods:The C-PC/CMC-NPs and C-PC/CMC-CD59sp-NPs were synthesized by the ionic cross-linking method(Ca Cl2 acted as crosslinking agent).The size potential of nanoparticles and zeta potential of C-PC,CMC and nanoparticles were determined using Malvern Nano ZS.The experiment was divided into four groups: control group,C-PC,C-PC/CMC-NPs and C-PC/CMC-CD59sp-NPs treatment group.The cytotoxic effects of drugs on cervical cancer He La and Si Ha cells were evaluated by the CCK8 cell viability assay.The effects of different drugs on cell cycle progress,cellular apoptosis cervical cancer He La and Si Ha cells were detected by flow cytometry.The effect of the phycocyanin uptake rate was examined by flow cytometry.Prove that C-PC/CMC-CD59sp-NPs is targeted.The effects of C-PC,C-PC/CMC and C-PC/CMC-CD59sp-NPs nanoparticles on cell apoptosis of cervical cancer He La and Si Ha cells were examed by TUNEL assay.We constructed the cervical cancer cells tumour mice model in NU/NU nude mice.Two to three weeks of administration using subcutaneous injections around the tumor mass.We further observed the tumor volume and tumor size to understand the antitumor effect in vivo.Immunofluorescence analysis and Western blot were used to quantify Bcl-2 and cleaved caspase-3 expression in tumor tissues.Western blot was further used to quantify CDK4,Cyclin D1,p21 and MMP-2expression in tumour rat model in vivo.Results:Results showed the morphology of the nanoparticles was well distributed with a diameter distribution of 200±11.3 nm and zeta potential of-19.5±4.12 m V.The zeta potential of C-PC was-18.4 ± 4.29 m V and the zeta potential of CMC was-40.4 ± 4.36 m V.The inhibitory effect of C-PC/CMC-CD59sp-NPs on cell proliferation were significantly higher than C-PC and C-PC/CMC-NPs.And the cell cycle arrest at G0/G1 phase were also significantly higher than C-PC and C-PC/CMC-NPs.By TUNEL method,it was found that targeting nanospheres to promote apoptosis was significantly higher than that of C-PC and C-PC/CMC-NPs treatment groups.C-PC/CMC-CD59sp-NPs specifically bind to cervical cancer cells through the targeting of CD59 sp,and the uptake of C-PC is significantly higher than that of C-PC and C-PC/CMC-NPs,prove that C-PC/CMC-CD59sp-NPs is targeted.By Western blot experiment,our study further found that C-PC/CMC-CD59sp-NPs can regulate the cell cycle by up-regulating the expression of p21 and then down-regulating the expression of Cyclin D1 and CDK4 in cervical cancer.Via up-regulating the expression of caspase-3 and down-regulating the expression of bcl-2 in cervical cancer,C-PC/CMC-CD59sp-NPs can promote the apoptosis of tumor cells and inhibit the growth of tumor by down-regulating the expression of MMP-2.Conclusion:Targeting nano-drug C-PC/CMC-CD59 sp exerts antitumor activity and its antitumor activity is mainly achieved by inhibiting the proliferation of cervical cancer cells,promoting cell apoptosis,inhibiting cell cycle and preventing the metastasis of tumor cells.Our research provides new ideas for the research and development of marine drugs and provides theoretical support for the targeted therapy of anticancer drugs.Provide theoretical basis and experimental basis for future clinical treatment of cervical cancer. |
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