| ObjectiveIn this paper,we prepared Herceptin-functionalized SK-BR-3 cell membrane-encapsulated paclitaxel(PTX)nanocrystals(HCNCs)for targeted therapy of HER2-positive breast cancer and investigated its antitumor effect.MethodsPaclitaxel nanocrystals(NCs)were prepared by a combination of anti-solvent precipitation and ultrasonic fragmentation.Cell membranes(CMs)were extracted from SK-BR-3 cells,and the amino groups on the cell membrane surface proteins were activated and linked with Herceptin(HCT)through amide bonds to obtain HCT-CMs.CMs and HCT-CMs were coated on NCs by ultrasonic incubation to obtain SK-BR-3 cell membrane-encapsulated paclitaxel nanocrystals(CNCs)and Herceptin-functionalized SK-BR-3 cell membrane-encapsulated paclitaxel nanocrystals(HCNCs).The HCNCs were characterized by transmission electron microscopy(TEM),X-ray diffraction(XRD),differential scanning calorimetry(DSC),sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE),in vitro release experiments,etc.The uptake of HCNCs by SK-BR-3 cells was investigated by cell uptake assay and flow cytometry.The targeting effect of HCNCs on HER2-positive breast cancer was verified by in vivo imaging experiments.The effects of HCNCs on SK-BR-3 cells were investigated by MTT assay,live/dead cell staining,flow cytometry and other apoptosis tests.The SK-BR-3 cell suspension was injected into the right breast fat pad of nude mice to establish a model,and the tumor inhibitory effect of HCNCs was studied in vivo.ResultsThe results of transmission electron microscopy(TEM)showed that the HCNCs had a cubic shape with a particle size of about 220 nm.The results of DSC and XRD indicated that PTX existed in the HCNCs in an amorphous state.In vitro release experiments showed that HCNCs had sustained release of PTX.Cell uptake experiments showed that the uptake ability of HCNCs by SK-BR-3 cells was significantly better than that of NCs and CNCs.In vivo imaging experiments demonstrated that HCNCs significantly accumulated in tumor tissues relative to NCs and CNCs,further confirming the good tumor targeting ability of HCNCs.Apoptosis experiments showed that HCNCs could significantly inhibit the proliferation of SK-BR-3 cells.In vivo tumor inhibition experiments demonstrated that HCNCs significantly inhibited tumor growth.ConclusionsIn this study,HCNCs were successfully prepared for the treatment of HER2-positive breast cancer.HCNCs have the advantages of dual targeting ability,long circulation time in vivo,less side effects,and high bioavailability.Therefore,HCNCs have good application prospects in the treatment of HER2-positive breast cancer. |
