ABSTRACT
Nanoparticular drug delivery system (NDDS) has great potential for enhancing the efficacy of traditional chemotherapeutic drugs. However, it is still a great challenge to fabricate a biocompatible NDDS with simple structure capable of optimizing therapeutic eï¬cacy, such as high tumor accumulation, suitable drug release profile (e.g. no premature drug leakage in normal physiological conditions while having a rapid release in cancer cells), low immunogenicity, as well as good biocompatibility. In this work, a simple core/shell structured nanoparticle was fabricated for prostate cancer treatment, in which a mesoporous silica nanoparticle core was applied as a container to high-efficiently encapsulate drugs (doxorubicin, DOX), CaCO3 interlayer was designed to act as sheddable pH-sensitive gatekeepers for controlling drug release, and cancer cell membrane wrapped outlayer could improve the colloid stability and tumor accumulation capacity. In vitro cell experiments demonstrated that the as-prepared nanovehicles (denoted as DOX/MSN@CaCO3@CM) could be efficiently uptaken by LNCaP-AI prostate cancer cells and even exhibited a better anti-tumor efficiency than free DOX. In addition, Live/Dead cell detection and apoptosis experiment demonstrated that MSN/DOX@CaCO3@CM could effectively induce apoptosis-related death in prostate cancer cells. In vivo antitumor results demonstrated that DOX/MSN@CaCO3@CM administration could remarkably suppress the tumor growth. Compared with other tedious approaches to optimize the therapeutic eï¬cacy, this study provides an effective drug targeting system only using naturally biomaterials for the treatment of prostate cancer, which might have great potential in clinic usage.
Subject(s)
Cell Membrane/metabolism , Doxorubicin/pharmacology , Drug Delivery Systems , Nanoparticles/administration & dosage , Prostatic Neoplasms/drug therapy , Silicon Dioxide/chemistry , Animals , Antibiotics, Antineoplastic/pharmacology , Apoptosis , Cell Proliferation , Drug Liberation , Humans , Hydrogen-Ion Concentration , Male , Mice , Mice, Inbred BALB C , Mice, Nude , Nanoparticles/chemistry , Porosity , Prostatic Neoplasms/enzymology , Prostatic Neoplasms/pathology , Tumor Cells, Cultured , Xenograft Model Antitumor AssaysABSTRACT
OBJECTIVE: To Construct the prokaryotic expression vector of the fusion gene IFN-alpha1b/CSP II. METHODS: IFN-alpha1b was amplified from the human genomic DNA by PCR and cloned into prokaryotic expression vector pGEX-4T-1. The recombinant plasmid pGEX-4T-1/IFN-alpha1b was constructed. Circumsporozoite protein II (CSP II) was amplified from the Plasmodium falciparum genomic DNA by PCR and was cloned into the prokaryotic expression vector pGEX-4T-1. The recombinant plasmid pGEX-4T-1/CSPII was constructed. IFN-alpha1b was cut from the recombinant plasmid pGEX-4T-1/IFN-alpha1b digested with BamH I and EcoR I and ligated with the recombinant plasmid pGEX-4T-1/CSP II also digested with BamH I and EcoR I. The recombinant prokaryotic plasmid pGEX-4T-1/IFN-alpha1b/CSP II was constructed. The fusion gene IFN-alpha1b/ CSP II was expressed in E. coli by IPTG. RESULTS: The prokaryotic expression vector pGEX-4T-1/IFN-alpha1b, pGEX-4T-1/CSP II and pGEX-4T-1/IFN-alpha1b/CSP II were identified by PCR, enzyme digestion and gene sequencing. The expressed fusion protein/IFN-alpha1b/CSP I in E. coli was identified by SDS-PAGE and Western blot. CONCLUSION: The prokaryotic expression vector of the fusion gene IFN-alpha1b/CSP II was successfully constructed, which was then expressed in E. coli.
Subject(s)
Genetic Vectors , Interferon-alpha/biosynthesis , Protozoan Proteins/biosynthesis , Recombinant Fusion Proteins/biosynthesis , Animals , Base Sequence , Cloning, Molecular , Escherichia coli/genetics , Escherichia coli/metabolism , Gene Expression , Interferon-alpha/genetics , Molecular Sequence Data , Plasmids/genetics , Polymerase Chain Reaction , Protozoan Proteins/geneticsABSTRACT
OBJECTIVE: To establish the computer-based video-tracking conditioned place preference (CPP) system in mice. METHODS: The CPP system was composed of computer, camera, soundproof box, shuttle box and analytical software. The results of morphine-induced conditioned place preference were used to evaluate the experiment system. And the effect of morphine-induced locomotor activity in drug-paired compartment was studied in mice. RESULTS: Low (1 mg/kg, i.p.), moderate (3 mg/kg, 5 mg/kg, i.p.) and high (10 mg/kg, i.p.) dose of morphine significantly prolonged the time mice spent in drug-paired compartment compared with saline, but there was no dose-response relation. Moderate and high dose of morphine significantly enhanced locomotor activity, among which 5 mg/kg and 10 mg/kg morphine induced behavior sensitization in drug-paired compartment during the conditioning sessions. CONCLUSION: The computer-based video-tracking conditioned place preference experiment system in mice established successfully is reliable and stable.
