JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES) ›› 2017, Vol. 55 ›› Issue (7): 43-48.doi: 10.6040/j.issn.1671-7554.0.2017.086

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Development and evaluation of chitosan gene nanoparticles loaded with recombinant plasmid pVAX1/Tat PTD-endostatin

ZHANG Xinke1, CHEN Hongyuan2, YANG Ying1, LI Yan1, WANG Fengshan1   

  1. 1. School of Pharmaceutical Sciences, Shandong University, Jinan 250012, Shandong, China;
    2. Department of General Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong, China
  • Received:2017-01-20 Online:2017-07-10 Published:2017-07-10

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Abstract: Objective To develop chitosan gene nanoparticles loaded with recombinant plasmid which contains endostatin(Es)gene and evaluate its in vitro inhibitory effects on HUVEC cells. Methods pVAX1 was taken as the expression vector. PCR combined with double restriction enzyme digest method was used to construct recombinant plasmids pVAX1/Tat PTD-Es. Chitosan nanoparticles loaded with plasmids were prepared by ion cross-linking method. Agarose gel electrophoresis experiment was employed to screen the best prescription of nanoparticles. The shape, Zeta potential and size distribution of nanoparticles were evaluated. After nanoparticles were transferred into HUVEC cells, ELISA method was used to detect the expression of Tat PTD-Es, and CCK-8 was explored to determine the inhibitory effect on HUVEC. In ELISA and CCK-8 experiments, LipofectamineTM2000 loaded with pVAX1/Tat PTD-Es was used as positive control. Results The recombinant plasmid pVAX1/Tat PTD-Es was constructed successfully. The Zeta potential of chitosan nanoparticles loaded with recombinant plasmid was(14.3±1.6)mV, the size distribution was(145±12)nm. The concentration of Tat PTD-Es in HUVEC supernatant was(182.5±16.3)ng/mL. CCK-8 results showed that chitosan nanoparticles loaded with pVAX1/Tat PTD-Es had inhibitory effects on HUVEC cells and the inhibitory rate 山 东 大 学 学 报 (医 学 版)55卷7期 -张新科,等.载pVAX1/Tat PTD-endostatin壳聚糖纳米基因载体的构建与体外活性评价 \=-in 72 h was(53.4±5.4)%(Z=-2.611, P=0.008). Conclusion Chitosan nanoparticles loaded with recombinant plasmid pVAX1/Tat PTD-Es can be transferred into HUVEC cells, and have distinct inhibitory effects on the proliferation of HUVEC cells.

Key words: Tat PTD-endostatin, Gene nanoparticle, Chitosan, Gene therapy

CLC Number: 

  • R318.08
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