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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2014, Vol. 34 Issue (8): 7-13    DOI: 10.13523/j.cb.20140802
    
Construction of the Eukaryotic Expression Vector of PhLTP Gene and Its Primary Antitumor Effects in vitro and in vivo
ZHAO Jing, LV Hui, Jiayinaguli·ZHUMABAI, SUN Su-rong
Xinjiang University College of Life Science and Technology, Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, Urumqi 830046, China
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Abstract  

Objective: To construct an eukaryotic expressing plasmid of lipid transfer protein from Peganum harmala, and investigate its inhibitory effects on the growth of murine melanoma(B16) cells in vitro and in vivo. Methods: The eukaryotic expressing plasmid of PhLTP(pcDNA3.1-PhLTP) was constructed by DNA recombination technique. The recombination plasmid and empty vector were transfected into B16 cells, respectively. The effect of pcDNA3.1-PhLTP on the growth of B16 cells was measured by MTT assay. B16 cells were inoculated subcutaneously of C57BL/6J mice, which randomly divided into 4 groups. Tumor volume and weigh were measured, and the tumor inhibition rates were calculated. The change of the mouse spleen and liver were observed under microscope. The expressions of PhLTP, vascular endothelial growth factor (VEGF) and basic fibroblast growth factor(bFGF) in tumor tissues were detected by immunohistochemistry. Results: The proliferation of B16 cells were inhibited after transfection with pcDNA3.1-PhLTP for 72 h(P< 0.01). Growth rate of tumor in mice model was also significantly inhibited in pcDNA3.1-PhLTP therapy group as compared with the control group and empty vector group (P< 0.05). The tumor tissue cells had different levels of necrosis in recombinant plasmid group. The liver, lungs and other organs in mice haven't significant pathological damage. The PhLTP could be expressed in recombinant plasmid group. And the positive expression indexes of VEGF and bFGF in recombinant plasmid group were significantly lower than ones of VEGF and bFGF in control group and empty vector group (P< 0.01). Conclusion: The recombinant eukaryotic expression vector pcDNA3.1-PhLTP is constructed successfully, and pcDNA3.1-PhLTP can inhibit the growth of B16 cells in vitro and in vivo. It is suggestted that the recombinant plasmid pcDNA3.1-PhLTP may have potential applications in the treatment of melanoma.

Key words： The gene of recombinant Peganum harmala lipid transfer protein (rPhLTP)      Transfection Growth inhibition      Murine melanoma cells (B16)      Angiogenesis     
Received: 21 May 2014      Published: 25 August 2014
ZTFLH:  Q255  
  R730.54  
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ZHAO Jing, LV Hui, Jiayinaguli·ZHUMABAI, SUN Su-rong. Construction of the Eukaryotic Expression Vector of PhLTP Gene and Its Primary Antitumor Effects in vitro and in vivo. China Biotechnology, 2014, 34(8): 7-13.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20140802     OR     https://manu60.magtech.com.cn/biotech/Y2014/V34/I8/7


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