title: A universal gene carrier platform for treatment of human prostatic carcinoma by p53 transfection.
abstract: Our previous work showed that a charge-reversal layer-by-layer nanosystem, PEI/PAH-Cit/AuNP-CS, effectively facilitates cellular uptake of siRNA and enhances the silencing efficacy of MDR1 siRNA. Here, the plasmid loading capacity of this vehicle was examined using EGFP-N1, and the plasmid release profile was determined in response to pH changes. The cytotoxicity of the EGFP-N1/PEI/PAH-Cit/AuNP-CS complex against HeLa and 293T cells was almost negligible. PEI/PAH-Cit/AuNP-CS efficaciously delivered the plasmids EGFP-N1 (encoding green fluorescent protein) and pGL3.0 (encoding luciferase) into 293T and HeLa cells, thus verifying the universality of PEI/PAH-Cit/AuNP-CS as a gene carrier. The results of an inverted fluorescence microscopy, flow cytometry (FCM) and western blotting methods demonstrated that PC-3 prostate cancer cells treated with EGFP-p53/PEI/PAH-Cit/AuNP-CS expressed higher levels of GFP than cells treated with EGFP-p53/PEI. Furthermore, PC-3 cells treated with EGFP-p53/PEI/PAH-Cit/AuNP-CS showed reduced cellular viability and increased nuclear fragmentation, consistent with elevated p53 expression. Propidium iodide (PI) flow cytometric assays were conducted to demonstrate that EGFP-p53/PEI/PAH-Cit/AuNP-CS elevated the level of apoptosis in PC-3 cells. Western blotting and caspase activation studies revealed that EGFP-p53/PEI/PAH-Cit/AuNP-CS complexes may induce PC-3 apoptosis via the mitochondria-mediated signaling pathway by up-regulation of Bax, down-regulation of Bcl-2, and activation of caspase-3.
authors.affiliation: Beijing Printed Electronics Engineering Technology Research Center, Beijing Institute of Graphic Communication, Beijing 102600, PR China; CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety, National Center for Nanoscience and Technology, Beijing 100190, PR China.
authors.forename: Lu
authors.initials: L
authors.lastname: Han
authors.affiliation: CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety, National Center for Nanoscience and Technology, Beijing 100190, PR China; Laboratory of Molecular Iron Metabolism, College of Life Science, Hebei Normal University, Shijiazhuang 050016, PR China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhao
authors.affiliation: CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety, National Center for Nanoscience and Technology, Beijing 100190, PR China.
authors.forename: Juan
authors.initials: J
authors.lastname: Liu
authors.affiliation: Laboratory of Molecular Iron Metabolism, College of Life Science, Hebei Normal University, Shijiazhuang 050016, PR China.
authors.forename: Xiang-Lin
authors.initials: XL
authors.lastname: Duan
authors.affiliation: Beijing Printed Electronics Engineering Technology Research Center, Beijing Institute of Graphic Communication, Beijing 102600, PR China.
authors.forename: Lu-Hai
authors.initials: LH
authors.lastname: Li
authors.affiliation: Beijing Printed Electronics Engineering Technology Research Center, Beijing Institute of Graphic Communication, Beijing 102600, PR China.
authors.forename: Xian-Fu
authors.initials: XF
authors.lastname: Wei
authors.affiliation: Beijing Printed Electronics Engineering Technology Research Center, Beijing Institute of Graphic Communication, Beijing 102600, PR China; Department of Chemistry and Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical Biology (Ministry of Education), Tsinghua University, Beijing 100084, PR China. Electronic address: weiyen@tsinghua.edu.cn.
authors.forename: Yen
authors.initials: Y
authors.lastname: Wei
authors.affiliation: CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety, National Center for Nanoscience and Technology, Beijing 100190, PR China. Electronic address: liangxj@nanoctr.cn.
authors.forename: Xing-Jie
authors.initials: XJ
authors.lastname: Liang
mesh_terms: D049109:Cell Proliferation; D002470:Cell Survival; D048271:Chitosan / Q000737:chemistry; D002949:Citraconic Anhydrides / Q000737:chemistry; D006046:Gold / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008297:Male; D053768:Metal Nanoparticles / Q000737:chemistry / Q000648:ultrastructure; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000378:metabolism; D011073:Polyamines / Q000737:chemistry; D011094:Polyethyleneimine / Q000737:chemistry; D011471:Prostatic Neoplasms / Q000235:genetics* / Q000473:pathology / Q000628:therapy*; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: D002949:Citraconic Anhydrides; D011073:Polyamines; D016159:Tumor Suppressor Protein p53; D051028:bcl-2-Associated X Protein; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C063994:polyallylamine; D006046:Gold; D011094:Polyethyleneimine; D048271:Chitosan
keywords: EGFP-p53 plasmid; PEI/PAH-Cit/AuNP-CS nanoparticles; Prostate cancer; Universality