A Novel Nanodrug Delivery System Based On Multifunctional Hollow Mesoporous Silica Nanoparticles Modified With Aptamer For The Target Co-Delivery Of Plasmid And Sorafenib And Its Application In Liver Cancer Therapy

Purpose:Cancer is still one of the most devastating diseases in the world,and the incidence and mortality of liver cancer are among the highest.Sorafine(S)is one of the effective drugs in the treatment of liver cancer,but there are still problems such as poor water solubility,low bioavailability and so on.Therefore,EGFR-sgRNA/Cas9 plasmid was designed and multifunctional targeted delivery system(SE@HM/P-aE)based hollow mesoporous silica nanoparticles modified with anti-EpCAM aptamer(aE)and PAMAM(P)was constructed for targeted co-delivery of sorafenib and plasmid to tumor cells in order to improve the sensitivity of tumor cells to sorafenib and effectively inhibit the growth of tumor.Method:First,synthesis conditions of HMSM were optimized and HMSN-COOH was prepared with suitable particle diameters.Then sorafenib was physically loaded into HMSN nanoparticles and P-aE was absorbed on the surface to constructe S@HIM/P-aE.The EGFR-sgRNA/Cas9 plasmid was constructed,then AO/EB staining and western-blot assay were used to verify its biological activities.EGFR-sgRNA/Cas9 plasmid was loaded into S@HM/P-aE by adsorption to form SE@HM/P-aE nanoparticles.TEM,DLS,AFM and IR were used to characterize SE@HM/P-aE.A standard curve of sorafenib was established by HPLC,the drug loading,entrapment efficiency and cumulative release of sorafenib were determined.The cytotoxicity of nanocarriers and nanodrugs on cells was detected by MTT assay.Finally,H22 cell transplantation model in Kunming mice was established to study tlhe anti-tumor effect in vivo.Hematoxylin-eosin(HE)staining was used to detect the side effects of nanodrugs.Immunohistochemistry(IHC)was used to explore mechanism that SE@HM/P-aE improved sensitivity of tumor cell to Sora.Results:SE@HM/P-aE nanoparticles are uniform in size(all below 200 nm)with pH response manner.The uptake assay indicated that NPs with the aptamer can significantly improve the uptake of the nanodrugs.In vitro biological experiments demonstrated that SE@HM/P-aE could enhance the synergistic inhibition of sorafenib and EGFR-sgRNA/Cas9 on tumor cell proliferation and promote cell apoptosis.In vivo experiments confirmed that SE@HM/P-aE had lower toxic and side effects,and could effectively inhibit tumor growth.Conclusion:A multifunctional targeted delivery system(SE@HM/P-aE)was successfully designed and constructed for co-delivery of sorafenib and EGFR-sgRNA/Cas9.SE@HM/P-aE could improve the water solubility and tumor-targeting property of sorafenib.SE@HM/P-aE could significantly increase the sensitivity of tumor cells to sorafenib.In vivo studies further confirmed the significant anti-tumor effect of SE@HM/P-aE,as well as low toxicity and side effects,and revealed the mechanism,providing new ideas for cancer treatment.