Anti-Angiogenic Activity of Naked Plasmid Encoding Targeting Recombinant Platelet Factor 4

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Abstract

To construct an recombinant eukaryotic expression vector of pcDNA3.1(+)-PF4^47-70-RGD, detect expression of targeting recombinant PF4 in eukaryotic cell and explore its biologic activity in vitro. A targeting recombinant PF4(platelet factor-4) eukaryotic expression vector was designed and constructed by cloning PF4^47-70-RGD cDNA into the pcDNA3.1(+) vector, and was transferred by lipofectamine into CHO cell line. Then G418 was used to select table-transfected CHO cell line with PF4^47-70-RGD gene. The mRNA and protein expression level of recombinant peptides were analyzed via RT-PCR and Western blot. MTT assay was used to observe the growth inhibition effect of expressing products of table-transfected CHO cell line on HUVEC(Human Umbilical Vein Endothelial Cells). The recombinant eukaryotic expression vector of pcDNA3.1(+)-PF4^47-70-RGD is successfully constructed and stabilized expressed CHO cell lines is acquired. Target gene expression is detected both at the genetic and protein level. MTT assay shows that growth inhibitory rate of expressing products of table-transfected CHO cell to HUVEC is 50.8%. Compared with the control group, the growth and tubular formation of HUVEC are significantly inhibited in the experimental group. The targeting recombinant PF4 peptides have antiangiogenic activity, and they inhibit the proliferation of HUVEC in vitro.

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	Articles by authors
	Wang Tingyue
	Wang Zhe
	Huang He

Keywords： platelet factor four； RGD peptide； anti-angiogenesis； cancer therapy； gene therapy

Received 2015-05-06;

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Wang Tingyue, Wang Zhe, Huang He.Anti-Angiogenic Activity of Naked Plasmid Encoding Targeting Recombinant Platelet Factor 4[J] Chemcial Industry and Engineering, 2017,V34(3): 88-94

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