H2A-functional Nanoparticles For Bortezomib/P53 Delivery And Apoptosis Induction

Malignant tumor is a major disease threatening global human health,and the number of new cases and deaths is increasing year by year due to environmental,population and lifestyle factors.It is estimated that the number of global cancer patients will increase from 17 million in 2018 to 26 million by 2040.More than 50%of the tumor patients had p53 mutations in their tissue samples.Therefore,in recent years,the design and preparation of drug/p53 gene co-delivery vector with real-time imaging function has become the focus of people's attention,which is expected to provide new safe and effective technical means for future tumor treatment.Based on the above considerations,in this paper,the uperconversion nanoparticle NaYF₄:Yb³⁺Er³⁺?UCNPs?was synthesized by high temperature solvent heat method,and then Tetraethyl orthosilide?TEOS?was wrapped to the surface of UCNPs nanoparticle by Stober method to form upconversion nanoparticle?UCNPs@mSiO₂?with uniform size and mesoporous silica.Finally,UCNPs@mSiO₂ was functionalized on the surface of histone H2A through APTES amino modification and EDC/NHS coupling reaction,and the organic-inorganic hybrid-upconversion nano-delivery vector UCNPs@mSiO₂-H2A was prepared.This vector can have a good bortezomib capacity and at the same time,due to the introduction of cationic histone H2A,it shows the characteristics that are conducive to cell uptake and effectively promotes the transfection of p53 gene.We used HeLa?wt-p53?and NCI-H1299?p53 null?as cell models to systematically evaluate the molecular mechanism of the vector mediating the co-delivery of bortezomib/p53 gene and inducing the apoptosis of tumor cells.After 48 hours of transfection,the apoptosis of NCI-H1299 cells was effectively enhanced due to the large expression of p53 gene.At the same time,the nano-carrier complex?UCNPs?BTZ?@mSiO₂-H2A/p53?can enhance the sensitivity of NCI-H1299 cells to the protease inhibitor bortezomib and effectively enhance the effect of inducing tumor cell apoptosis through the expression of exogenous genes.Finally,molecular mechanism studies showed that after UCNPs@mSiO₂-H2A mediated p53 transfection,the expression levels of pro-capspase 3 and pro-caspase 9proteins decreased,and the activities of caspase 3 and caspase 9 were enhanced,indicating that the vector system enhanced the apoptosis of tumor cells through the activation of caspase 3 and caspase 9.At the same time,due to the decreased Bcl-2/Bax protein expression ratio and the down-regulation of mitochondrial membrane potential,it was proved that the carrier mediated p53 delivery could activate the mitochondrial apoptosis pathway and thus effectively inhibit the proliferation of tumor cells.In summary,the organic-inorganic hybrid complex synthesized by functional modification of histone H2A as a new carrier can effectively achieve the co-delivery of drugs and genes,through the expression of the target gene in tumor cells,therapeutic proteins are produced,which effectively improve the sensitivity of the target gene to proteasome inhibitors and greatly enhance the inhibition and killing ability of malignant tumor cells,thus improving the therapeutic effect.Overall,this research not only provides a new design for inorganic up-conversion nanoparticles in the field of gene therapy,but also the carrier can achieve the co-delivery of drugs and genes,laying a foundation for the research and clinical application of gene therapy for malignant tumors good foundation.