Targeted Inhibition Of Hepatocellular Carcinoma By Membrane Loaded Natural Product Hypocrellin B Nanovesicles And Its Mechanism

Hepatocellular carcinoma(HCC)is one of the most common malignancies in the world,with high morbidity and mortality.At present,there are mainly radical resection,liver transplantation,radiotherapy and chemotherapy in the clinical treatment of HCC,but the clinical treatment effect is not satisfactory.Existing treatment methods mainly have defects such as large surgical trauma,easy to develop tumor resistance,and poor prognosis.New therapies for hepatocellular carcinoma are urgently needed.Photodynamic therapy(PDT),with its unique advantages of minimally invasive,good selectivity,high safety and unique efficacy,has been used as a new therapeutic method for tumor treatment.The process of PDT treatment of tumor mainly depends on the specific wavelength of photosensitizer,which reacts with the oxygen in the tumor site to produce a large number of reactive oxygen species(ROS).As a natural product,Hypocrellin B has excellent photosensitive effect and can be used as a photosensitive agent for PDT treatment of many diseases,especially for tumor treatment.Due to its poor water solubility,fast metabolism in vivo and no specific tissue distribution,these characteristics have become the main obstacles for the application of HB as a photosensitizer in PDT.With the development of nanotechnology,nano-drug delivery system can improve drug solubility,improve drug stability,increase drug targeting,reduce drug toxicity and side effects and improve the adverse effects of photosensitizers.Systematic administration of nanomedicine delivery agents may activate the rapid elimination of autoimmunity,and the uncleared nanomedicine is difficult to penetrate various biological barriers,leading to little clinical benefit of this system.Endogenous cells have the advantages of avoiding immune clearance,strong biocompatibility and long blood circulation time.According to the advantages of endogenous cells,encapsulating nanoparticles in cell membranes can make up for the deficiency of immune clearance in the delivery system of nano-drugs,thus improving the biological distribution of drugs,increasing the circulation time of drugs,and thus increasing the therapeutic effect of drugs.Based on the relationship between tumor pathogenesis and endogenous cells,this paper aims to develop bionic endogenous cells with photosensitizer nanoparticles loaded on cell membranes,so as to enhance the anti-tumor therapeutic effect of photosensitizer synergistic PDT.The main content of this study is divided into the following parts:1.Tumor-targeted ligand modification of homologous hepatocellular carcinoma cell membrane with arthromycin nanoparticles to enhance the anti-hepatocellular carcinoma effect of PDT.First,homologous liver cancer cells were collected by membrane extraction method to collect hepatoma cell membrane(CCCM),and tumor-targeting ligand TF was modified into homologous liver cancer cell membrane(TF-CCCM)by maleamide biochemical reaction.Ethylene nanoparticles(HB NPS)were prepared by double emulsion method.TF-CCCM-HB NPS was prepared by extrusion method by mixing the two in a certain proportion.TEM observation showed that the nanoparticles had regular morphology and a membrane of about 10nm around HB NPS.TF-CCCM-HB NPS showed good stability in vitro and retained the protein components of homologous liver cancer cell membrane.TF-CCCM-HB NPS has strong targeting ability and phagocytosis in homologous hepatoma cells,and inhibits the growth of tumor cells in a dose-dependent manner.TF-CCCM-HB NPS synergistic with PDT can promote the production of ROS in cancer cells and reduce mitochondrial membrane potential.TF-CCCM-HB NPS has strong tumor targeting and retention effect in mice with hepatocellular carcinoma.TF-CCCM-HB NPS combined with PDT can achieve partial tumor ablation in mice,prolong the survival time of tumor-bearing mice,and inhibit tumor metastasis and recurrence.TF-CCCM-HB NPS has good biosafety,indicating that TF-CCCM can be used as a targeted delivery system of photosensitizer drugs to enhance the efficacy of PDT in the treatment of tumors.2.Targeting enhancement of PDT against hepatocellular carcinoma by neutrophil membrane-loaded arthromycin nanoparticles and its mechanism.Mice were intraperitoneally injected with LPS to replicate the classic model of mouse inflammation,and whole blood was collected to isolate neutrophils from peripheral blood of mice.Flow cytometry was used to detect the receptors on the surface of neutrophils,and the neutrophils were confirmed to be activated and the neutrophils were collected.Ethyl nanoparticles were prepared by double emulsion method.NM-HB NPS was prepared by extrusion method by mixing the two in a certain proportion.The structure of NM-HB NPS was determined by TEM,UV-Vis and FL,and the preparation parameters were optimized.In vivo and in vitro experiments have shown that nm-HB NPS has good biosecety and strong tumor targeting ability,which can increase the retention time of tumor site,inhibit tumor growth of hepatocellular carcinoma tumor-bearing mice,and improve the survival time of mice.In addition,Western blotting and RT-PCR were used to confirm the anti-liver cancer effect of NM-HB NPS synergistic with PDT in regulating JunB/ROS/mitochondrial apoptosis signaling pathway.