专业外语教学（吉林大学）140710-Supporting information.docVIP

Combination of doxorubicin-based chemotherapy and polyethylenimine/p53 gene therapy for the treatment of lung cancer using porous PLGA microparticles Xiaozheng Shi a,c, Chunjie Li a,c, Lingfei Zhang a,c, Sai Gao a,c, Haobo Han a,c, Jianxu Zhang a,c, Wei Shi a,b,c,*, Quanshun Li a,b,c,* a Key Laboratory for Molecular Enzymology and Engineering, the Ministry of Education, Jilin University, Changchun 130012, China b National Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun 130012, China c School of Life Sciences, Jilin University, Changchun 130012, China *Corresponding author. Tel.: +86-431 Fax: +86-431 E-mail address: quanshun@jlu.edu.cn (Q. Li); shiw@jlu.edu.cn (W. Shi). Materials and methods 1. Preparation and characterization of PEI25K/p53 nanoparticles PEI25K was dissolved in distilled water at a concentration of 1 mg/mL and adjusted to pH 7.4. The nanoparticles were prepared by gently mixing PEI25K and the plasmid p3XFLAG-CMV-p53 together at different mass ratios, and then incubated at room temperature for 30 min before use. The gel retardation assay was conducted to analyze the ability to condense the plasmid DNA, using 1% agarose gel electrophoresis in TAE buffer solution (90 V, 45 min). The mean particle size and zeta potential values were determined by dynamic light scattering (Malvern Zetasizer Nano ZS90, Malvern, UK) at room temperature. 2. Synergistic cytotoxicity of doxorubicin and p53 in A549 cells The A549 cells were seeded onto 96-well plates at a density of 1.0×104 cells per well, pre-incubated for 24 h in fresh serum-free DMEM medium, and further incubated with a mixture solution of plasmid p3XFLAG-CMV-p53 (final concentrations of 0-1000 ng/mL) using PEI25K as transfection agent and doxorubicin (0.1, 1 or 10 μg/mL). After 24 h, 20 μL of MTT solution (5 mg/mL in PBS) was added to each well, and the plate was incubated for an additional 4 h. Afterwards, the MTT solution was removed from each well, and 20