Preparation And Antitumor Effect Of Graphene Nano Drug-loading System

Graphene(GR)is a two-dimensional atomic crystal composed of carbon atoms mixed in the form of sp2.It has a large surface area,good biocompatibility,easy solubility,high drug carrying capacity,and easy cell membrane penetration.characteristic.In recent years,due to the trend of cancer patients increasing year by year,it is urgent to solve the strong side effects of radiotherapy and chemotherapy drugs in the treatment of cancer.For this reason,nano graphene oxide(NGO)was prepared by using graphene as a carrier.The diol(PEG)and polyethyleneimine(bPEI)modified nano-graphene oxide loaded anticancer drug cisplatin(CDDP)explored the cellular targeting and antitumor effects of nano drug carriers.The specific research scheme is as follows:(1)Graphene oxide(GO)was prepared by chemical oxidation-reduction method.Single-layer nano-graphene oxide(NGO)was obtained by ultrasonic fragmentation and peeling.The nano-flakes of graphene oxide were characterized by scanning electron microscopy(SEM).By modifying NGO with polyethylene glycol(PEG),graphene oxide nanoparticles with excellent water solubility and biocompatibility were obtained,which were loaded with cisplatin(CDDP)through covalent bonds.The ultraviolet-visible(UV-Vis)and Fourier-infrared spectroscopy(FTIR)of NGO-PEG-CDDP further indicate that cisplatin(CDDP)has been successfully loaded on graphene oxide nanosheets.By calculating the loading rate of NGO-PEG on CDDP,42.4% of CDDP was combined with NGO-PEG.(2)Under certain conditions,branched polyethyleneimine(bPEI)was grafted onto the graphene oxide nanocomposite to form NGO-PEG-bPEI-CDDP.The NGO-success was successfully prepared by UV-Vis and FTIR.The PEG-bPEI-CDDP nano drug-loading system measured the drug loading rate of CDDP by NGO-PEG-bPEI to 34.6%.(3)The antibodies in the serum of different tumor patients were separated and extracted by low-temperature ethanol method.The tumor-specific antibodies isolated and extracted by ELISA were highly specific.The results of polyacrylamide gel electrophoresis(SDS-PAGE)demonstrated that the NGO-PEG-bPEI-CDDP nano drug-loading system strongly adsorbed the extracted tumor antibody protein,and NGO-PEG-bPEI-CDDP-Antibody nano drug was obtained system.(4)MTT targeting experiments,enzyme-linked immunosorbent assay(ELISA)and immunofluorescence labeling assays were used to detect the targeting effect of the prepared NGO-PEG-bPEI-CDDP-Antibody nano drug-loading system on tumor cells,as follows:(1)MTT targeting experiment and enzyme-linked immunosorbent assay(ELISA)demonstrated that The NGO-PEG-bPEI-CDDP-Antibody nano-drug delivery system combined with antibodies showed a targeting effect on human nasopharyngeal carcinoma cells(CNE-1)and human oral squamous cell carcinoma cells(KB),and the targeting effect on CNE-1 cells was stronger than that on KB cells.The NGO-PEG-bPEI-CDDP-Antibody nano-drug delivery system linked with specific antibodies extracted from nasopharyngeal carcinoma serum was effective for KB cells and CNE-1 cells.The nano-drug delivery system of NGO-PEG-bPEI-CDDP-Antibody,which links specific antibodies extracted from tongue cancer serum,has the strongest targeting effect,followed by the nano-drug delivery system of NGO-PEG-bPEI-CDDP-Antibody,which links specific antibodies extracted from hypopharyngeal cancer serum.(2)Immunofluorescence labeling experiments observed NGO-PEG-bPEI-CDDP-Antibody nano drug delivery system It can be enriched on the cell membrane surface of KB cells and CNE-1 cells,showing green fluorescence,but has no targeting effect on BV-2 cells,and there is no green fluorescence on the cell membrane surface.Moreover,the targeting effect of NGO-PEG-bPEI-CDDP nano drug-loading system on CNE-1 cells is stronger than that of KB cells.The experiment further confirmed that NGO-PEG-bPEI-CDDP-Antibody nano drug-loading system for tumor cells Targeted,no targeting to normal cells.(5)Study on the killing effect of NGO-PEG-bPEI-CDDP-Antibody nano drug-loading system on tumor cells:(1)The killing effect of NGO-PEG-bPEI-CDDP-Antibody nano drug-loading system on KB and CNE-1 cells at the same dose concentration and the same action time was better than that of NGO-PEG-CDDP and NGO-PEG-bPEI-CDDP.The killing effect is better,and the half-lethal dose of the cells is lower.This is because after the tumor-specific antibody is linked,the NGO-PEG-bPEI-CDDP-Antibody nano drug-loading system can directly bind to the antigenic polypeptide secreted on the surface of the tumor cell,and the killing time is shorter and the effect is better.(2)NGO-PEG-bPEI-CDDP-Antibody nano drug-loading system as a carrier of anti-cancer drug CDDP can effectively load CDDP,NGO-PEG-bPEI-CDDP-Antibody nano drug-loading system has multiple killing effects on cancer cells,Reducing the dose of CDDP can also enhance the killing effect on cancer cells.At the same dose concentration and the same action time,the killing effect of NGO-PEG-bPEI-CDDP-Antibody nano drug-loading system on KB and CNE-1 cells was better than that of NGO and CDDP alone.This is because the modification of PEG and bPEI increases the killing effect of CDDP,and the toxicity of NGO itself also increases the antitumor effect of the drug-loading system.