| Choriocarcinoma referred to as choriocarcinoma,which is a kind of highly malignant tumor.It is secondary to hydatidiform mole,abortion or term delivery,and a few can occur after ectopic pregnancy.Treatment of choriocarcinoma with systemic chemotherapy,supplemented by surgery with huge side affects.Therefore,the development of targeted chemotherapy drugs can reduce the dose and treatment time in drug treatment of choriocarcinoma.It will greatly reduce the side effects of drugs on other organs and tissues,and has important economic and social benefits to improve the life quality of patients and reduce the occurrence of harmful complications.Prodigiosin(PG)has antitumor,anti bacterial,anti fungal,anti malarial and immunosuppressive activities.The research group screened a prodigiosin producing strain named HDZK-BYSB107 from the natural anti termite Chamaecyparis lawsoniana.Previous experimental studies showed that the strain HDZK-BYSB107 has the effect of inhibiting proliferation and inducing apoptosis of many kinds of cancer cells,but the toxicity to normal cells is very small.The subject combination unit had been obtained choriocarcinoma cell targeting marker peptides VAR2 CSA.The Poly(lysine)dendrimer(DGL)has good biocompatibility and biodegradability with no toxic side effects.It has been widely used as a drug delivery carrier in the field of biomedicine.Therefore,in this study,strain HDZK-BYSB107 was used as model drug and DGL was served as a model drug delivery carrier for the preparation of targeting peptides VAR2 CSA choriocarcinoma endophyte prodigiosin DGL nano medicament.In order to achieve the purpose of targeted therapy of choriocarcinoma;at the same time,this study explored the DGL-PEG-CSA/PG NPs anticancer mechanism.The optimum preparation conditions of DGL-PEG/PG NPs,DGL-PEG-CSA/PG NPs and DGL-PEG-SCR/PG NPs were determined.At the same time,the biological safety,drug release in vitro and in vivo pesticide effect were evaluated.Test results show that:1.Purification and identification of strain HDZK-BYSB107 prodigiosin(PG),and the anti choriocarcinoma test results showed that the strain HDZK-BYSB107 PG has strong killing effect on the choriocarcinoma cell line(JEG3)and has minimal side effects or no obvious growth inhibition on human renal epithelial cell line(293T).2.The resulting DGL,DGL-PEG-CSA NPs and DGL-PEG-CSA/PG NPs with round shape,smooth surface suitable particle size and good dispersion.The average particle size was 70.68±11.26 nm,101.00±16.49 nm,786.96±37.93 nm,Zeta potential were?(10.32±0.68)m V,-(5.54±0.46)m V and-(6.94 + 0.65)m V respectively,the encapsulation rate of DGL-PEG-CSA/PG NPs were(89.39 + 1.83)% and drug loading capcity were(41.36 + 0.87)% respectively.3.The results of in vitro cell uptake test showed that the peptide VAR2 CSA was successfully attached to DGL and maintained a good biological activity,and could be targeted to JEG3.4.The results of cytotoxicity test showed that the survival rates of 293 T cells treated with DGL(5?g/ml,10?g/ml,20?g/ml,50?g/ml and 100?g/ml)were all very high.When the concentration of DGL were 100?g/ml and the survival rate of 293 T cells were(81.62±0.83)%,which indicates that DGL has good biocompatibility and no obvious side effects.5.The results of in vitro release test showed that DGL-PEG-CSA/PG NPs had a slow release property in p H=7.4,and the release amount of PG was higher than that of p H=7.4 at p H=5.3.It can be circulating well in the blood.6.The in vivo efficacy test results showed that DGL-PEG-CSA/PG NPs treated group has the minimum tumor weight(0.39±0.03 g)and the tumor volume was always less than 200 cm3 until 18 d.The survival curve showed that the DGL-PEG-CSA/PG NPs treated JEG3 beared nude mice has the highest survival level and with no side effect compared to other groups.We can infer that DGL-PEG-CSA/PG NPs has the best treatment effect.Additionally,this results showed that the peptide VAR2 CSA can specifically target JEG3 cells.7.The anticancer mechanism study showed that DGL-PEG-CSA/PG NPs can induce the expression of caspase-3,caspase-9,bax,P53 and AIF,PARP1 proteins up-regulated,while the expression of Bcl-2,XIAP,c IAP1 and c IAP2 proteins were down-regulated.The expression of caspase-3 and P53 proteins in JEG3 cells treated with caspase-3 inhibitor and P53 inhibitor did not significantly up-regulated.In this study,we successfully constructed a delivery system with HDZK-BYSB107 PG as a model drug and the poly(lysine)dendrimer was used as the drug carrier,which can not only remain the activity of polypeptide VAR2 CSA,but also realize the purpose of drug slowly release in vivo.This method achieves the purpose of safe,efficient,fast,low toxic and without side effects in the targeting treatment of choriocarcinoma.Therefore,the study of choriocarcinoma targeted therapy provides a new way of thinking and provide a strong theoretical basis for the clinical application of endophytic bacteria HDZK-BYSB107 prodigiosin. |
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