Chemoenzymatic Synthesis Of Cholesterol-g-Poly(amine-co-ester) Amphiphilic Copolymer As A Carrier For P53 Delivery

In all over the world,the incidence of cancer and mortality have shown an increasing trend year by year,making it a threat to human health and an urgent need to resolve the malignant disease.In recent years,the use of gene therapy to deliver specific genes and promote the expression of desired therapeutic protein has become the focus of attention in the study of tumor diseases,and it is expected to provide a new safe and effective strategy for future cancer treatment.Based on the above considerations,the lipase-catalyzed polymerization of N-methyldiethanolamine,diethyl sebacate and ?-pentadecanolide was performed to construct a cationic poly(amine-co-ester),and a hydrophobic N-(2-bromoethyl)-carbamoyl cholesterol was then grafted onto its main chain through a quaternization reaction to prepare the amphiphilic copolymer Chol-g-PMSC-PPDL.It has been verified that the novel carrier can well assemble with DNA to form a stable nanocomposite.Due to the introduction of hydrophobic fragment,the carrier showed the characteristics of facilitating cellular uptake and efficiently improving the transfection efficiency of p53 gene.Then the inhibiton of cell proliferation,migration and invasion of tumor cells after p53 transfection was evaluated using human cervical cancer cell line HeLa(p53 wild-type)and human prostate cancer cell line PC-3(p53 deletion type)as models.After 48 h,these two cells showed obvious inhibition of cell proliferation owing to the expression of therapeutic p53 protein.Meanwhile,the Chol-g-PMSC-PPDL/p53 transfection could induce the cell apoptosis and cell cycle arrest of HeLa and PC-3 cells.In addition,the carrier-mediated p53 delivery could suppress the migration and invasion of tumor cells and thus effectively prevent the spreading of cancer cells.Finally,the molecular meahnism analysis indicated that the expression levels of pro-caspase-3 and pro-caspase-9 have been detected to de dcreased after p53 transfection indicating the activation of these two apoptosis-associated proteins.Additionally,the decreased mitochondrial membrane potential after p53 transfection demonstrated that the carrier-mediated p53 delivery could activate the cell apoptosis in the mitochondria dependent apoptosis signaling pathway.In summary,cholesterol-g-poly(amine-co-ester)could be successfully synthesized in a chemoenzymatic route and used as a carrier for p53 gene delivery.Through the obvious expression of exogeneous therapetical protein,the inhibition and killing efficacy could be clearly observed.Overall,this study not only provided a green route for constructing biocompatible poly(amine-co-ester)derivatives and also facilitating the basic and application of gene therapy through the precise delivery of therapeutic genes.