Treatment Of Pancreatic Cancer With Doxorubincin NGR-PEG-PE Nanoparticles

Objective:Pancreatic cancer is the one of the most difficult to overcome cancer for the medical profession,the molecular targeted therapy is the most popular of the current research.In our research,Aminopeptidase N as a target,we produced NGR peptide modified Doxorubicin-PEG-PE micelles.And assay of its drug loading,encapsulation efficiency and nano particle diameter.We also study the inhibition of the human pancreatic cancer cells SW1990 and NGR head-modified Doxorubicin micelles(DOX-PEG-PE)for therapeutic effect of pancreatic cancer in nude mouse.Method:1.To prepare Doxorubicin-PEG-PE micelles:by PEG2000-DSPE and Mal-PEG3500-DSPE,film dispersion method to envelop Doxorubicin,hydration to micelles.2.Use of polymerization to connect NGR peptide and Doxorubicin micelles(DOX-PEG-PE).To study the micelles size,drug loading,encapsulation efficiency by laser particle size analyzer and high performance liquid chromatography.3.In vitro test:(1)First,transfected human pancreatic cancer cells SW1990 with the Luciferase transgenic method.The cell inhibition ratios assay(MTT assay)that was measured the same concentrations and the same time of Doxorubincin?Doxorubicin-PEG-PE and NGR peptide-modified doxorubicin micelle(NGR-DOX-PEG-PE).(2)By Flow cytometry and Confocal to evaluation the ability of get into the cell among Doxorubincin?Doxorubicin-PEG-PE and NGR peptide-modified doxorubicin micelle.4.Establishment of orthotopic model of pancreatic carcinoma:Use of amobarbital anesthesia by tail vein injection method,opened 1 cm's cut by the side of left upper quadrant of rectum,carefully separated stamoch and spleen and exposed pancreas,then used insulin syringes to injected cell suspension of a concentration of 5×106 ?/ml at a portal into the head of the pancreas near the spleen.And then,put pancreas back to the abdominal cavity,and stitched skin with 6-0 surgical suture.The nude mice were no death and wound infection happened,then the pancreatic cancer model was completed.5.In vivo anticancer assay:Established orthotopic model of pancreatic cancer were randomly divided into four groups,including saline group?Doxorubicin group?Doxorubicin-PEG-PE group and NGR peptide-modified doxorubicin micelle group.We gave corresponding drug to each group every three days.The nude mice were sacrificed after injecting drug three times,and then observation the sizes of tumor scales.Results:The drug loading and encapsulation efficiency were 10.98%and 97.62%,micelle particle size of 15-20nm.The results of in vitro test:NGR peptide-modified doxorubicin micelle was significantly stronger than the rest of the group of the ability to enter cells.The study of in vivo anticancer assay:NGR peptide-modified doxorubicin micelle was smallest than the rest of the group of the tumor volumes(P<0.05).Conclusion:Aminopeptidase N as a target,NGR peptide-modified doxorubicin micelle have the best inhibition of the growth of panceatic cancer,and the side effects are smallest.Provides a broader prospect for molecular targeted therapy of pancreatic cancer.