Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (20): 3670-3674.doi: 10.3969/j.issn.1673-8225.2011.20.015

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Construction of lentiviral vector of endothelial lipase and expression in infected HepG2 cells

Xu Bo, Deng Yu-bin   

  1. Department of Pathology and Pathophysiology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou  510080, Guangdong Province, China
  • Received:2010-12-29 Revised:2011-03-23 Online:2011-05-14 Published:2011-05-14
  • Contact: Deng Yu-bin, Doctor, Professor, Department of Pathology and Pathophysiology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, Guangdong Province, China dengyub@mail.sysu.edu.cn
  • About author:Xu Bo★, Studying for master’s degree, Department of Pathology and Pathophysiology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, Guangdong Province, China aquila851@yahoo. com.cn
  • Supported by:

    the Doctoral Foundation of Ministry of Education of China, No.20090171110048*

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Abstract:

BACKGROUND: Endothelial lipase expresses mainly in endothelial cells, and plays an important role in atherosclerosis and other diseases. But the mechanism is not very clear.
OBJECTIVE: To construct a lentiviral vector carrying rat endothelial lipase (EL) gene to transfect HepG2 cells.
METHODS: 1 482 bp DNA fragment of rat EL was amplified by PCR, cleaved with SpeⅠ, EcoRⅠand cloned into the lentiviral vector PRRL.sin.CMV.eGFP to construct PRRL.sin.CMV.EL-eGFP vector. 293FT cells were co-transfected with recombined PRRL.sin.CMV.EL-eGFP vector and packaging plasmids to produce PRRL.sin.CMV.EL-eGFP lentiviral viruses. Viruses were used to infect HepG2 cells. RT-PCR assay and western blot were used respectively to evaluate the mRNA and protein expressions of EL in the transfected HepG2 cells.
RESULTS AND CONCLUSION: The recombinant inducible lentiviral vector containing EL gene was successfully constructed. The lentiviruses produced by 293FT were efficient to transfect HepG2 cells. The mRNA and protein expressions of EL in the infected HepG2 were obviously up-regulated. It is indicated that EL expression in HepG2 cells can be effectively and stably up-regulated by infecting with the recombined PRRL.sin.CMV.EL-eGFP lentivirus.

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