title: Induction of apoptosis in cancer cells through N-acetyl-l-leucine-modified polyethylenimine-mediated p53 gene delivery.
abstract: Herein, N-acetyl-L-leucine-modified polyethylenimine was successfully constructed through the EDC/NHS-mediated coupling reaction and employed as vectors to accomplish p53 gene delivery using HeLa (p53wt) and PC-3 cells (p53null) as models. Compared with PEI25K, the derivatives exhibited lower cytotoxicity, protein adsorption and hemolytic activity, together with satisfactory pDNA condensation capability and gene transfection efficiency. After p53 transfection, MTT analysis confirmed that the cell proliferation was inhibited. Flow cytometric analysis showed that the derivative-mediated p53 delivery could induce stronger early apoptosis than PEI25K and Lipofectamine(2000). Further, PC-3 cells showed higher sensitivity to the exogenous p53 transfection than HeLa cells. The mechanism for inducing apoptosis was determined to be up-regulation of p53 expression at both mRNA and protein levels using RT-PCR and western blotting analysis. Expression level and activity analysis of caspase-3, -8 and -9, and mitochondrial membrane potential measurement revealed that p53 transfection mediated by these derivatives facilitated early apoptosis of tumor cells via a mitochondria-dependent apoptosis pathway. Thus, the derivatives showed potential as biocompatible carriers for realizing effective tumor gene therapy.
authors: Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, School of Life Sciences, Jilin University, Changchun 130012, China.
authors: Zhiyuan
authors: Z
authors: Li
authors: Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, School of Life Sciences, Jilin University, Changchun 130012, China.
authors: Liu
authors: L
authors: Zhang
authors: Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, School of Life Sciences, Jilin University, Changchun 130012, China. Electronic address: quanshun@jlu.edu.cn.
authors: Quanshun
authors: Q
authors: Li
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D020169:Caspases / Q000096:biosynthesis; D045744:Cell Line, Tumor; D018014:Gene Transfer Techniques / Q000009:adverse effects; D016158:Genes, p53; D006367:HeLa Cells; D006801:Humans; D007930:Leucine / Q000009:adverse effects / Q000031:analogs & derivatives* / Q000737:chemistry; D053078:Membrane Potential, Mitochondrial; D011094:Polyethyleneimine / Q000009:adverse effects / Q000737:chemistry*; D014162:Transfection
chemical_list: D011094:Polyethyleneimine; D020169:Caspases; D007930:Leucine; C088117:acetylleucine
keywords: Apoptosis; Biocompatibility; N-acetyl-l-leucine; Polyethylenimine; Transfection; p53 Gene