Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (15): 2737-2740.doi: 10.3969/j.issn.1673-8225.2012.15.017

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Construction of a microRNA expressing eukaryotic vector targeting vascular endothelial growth factor  

Xiong Jian-ning1, 2, Sun Jian2, Ou Ying-liang2, Jian Zhi-xiang2   

  1. 1Southern Medical University, Guangzhou  510080, Guangdong Province, China; 2Department of Hepatobiliary Surgery, Guangdong Provincial People’s Hospital, Guangzhou  510080, Guangdong Province, China
  • Received:2011-11-03 Revised:2012-03-08 Online:2012-04-08 Published:2012-04-08
  • Contact: author: Sun Jian, Doctor, Associate chief physician, Associate professor, Department of Hepatobiliary Surgery, Guangdong Provincial People’s Hospital, Guangzhou 510080, Guangdong Province, China akeyman@yahoo.com.cn
  • About author:Xiong Jian-ning★, Studying for master’s degree, Attending physician, Southern Medical University, Guangzhou 510080, Guangdong Province, China; Department of Hepatobiliary Surgery, Guangdong Provincial People’s Hospital, Guangzhou 510080, Guangdong Province, China neilxjn@163.com
  • Supported by:

    the Science and Technology Program of Guangdong Province, No. 2009B080701021*, 2010B080701021*; the Medical Science Research Foundation of Guangdong Province, No. A2010002*

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Abstract:

BACKGROUND: Studies have shown that the microRNA (miRNA) and its potential target gene play an important role in hepatocellular carcinoma, but the exact mechanism is still unclear.
OBJECTIVE: To construct a vascular endothelial growth factor (VEGF) miRNA expressing eukaryotic vector and to identify biological activity of VEGF miRNA transfected into HepG-2 cells.
METHODS: According to the sequence of VEGF mRNA, the VEGF miRNA was designed and synthesized, and then cloned into the pcDNA6.2-GW/EmGFP-miRNA vector and transfected into HepG-2 cell lines. The integrity of the insert fragment was detected using colony PCR and sequencing analysis. The biological activity of VEGF miRNA by way of real-time PCR and Western blot was determined. 
RESULTS AND CONCLUSION: Sequences of the inset fragment in the four miRNA expressing recombinants were correct. VEGF mRNA expression of the four miRNA recombinants were significantly decreased (P < 0.05), especially in the VEGF-miRNA-3 with an inhibitory rate of 87%. Four VEGF-targeting miRNA expressing recombinants were successfully constructed which can significantly inhibit VEGF gene expression in HepG2 cells.
Xiong JN, Sun J, Ou YL, Jian ZX. Construction of a microRNA expressing eukaryotic vector targeting vascular endothelial growth factor. Zhongguo Zuzhi Gongcheng Yanjiu. 2012;16(15): 2737-2740.     [http://www.crter.cn  http://en.zglckf.com]
 

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