Basic & Clinical Medicine ›› 2013, Vol. 33 ›› Issue (4): 418-422.

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Construction of CHD5 gene lentiviral vector and its expression in colorectal cancer cells

  

  • Received:2012-01-17 Revised:2012-06-12 Online:2013-04-05 Published:2013-03-15
  • Supported by:
    the National Natrual Science Foundation for young scholars of China

Abstract: Objective To construct the recombinant lentiviral vector containing human CHD5 gene and measure the expression level of CHD5 in LOVO cell. Methods The CHD5 fragment was amplified by PCR and cloned into the lentiviral vetor TREAutoR3.The lentiviral voter TREAutoR3 CHD5 was identified by restriction enzyme and DNA sequencing. Recombinant lentiviral was produced by 293FT cells following co-transfection of TREAutoR3 CHD5 with the packaging plasmids pCMV-VSV-G, pRSV-Rev and pMDLg-pRRE. Human Colorectal cancer LOVO cell were infected by the recombinant lventiviruse. The expression of CHD5 was confirmed by RT-PCR and Western blotting, and the proliferation of LOVO cells was evaluated by MTT assay. Results The recombinant lentiviral vecor carried the CHD5 gene was successfully constructed. RT-PCR and western blot analysis revealed that the CHD5 can be correct transcript and translated in human colorectal cancer LOVO cell which infected by the recombinant lentiviral lenti CHD5. After infected, the growth of LOVO cell is inhibited obviously. Conclusion It can be deliver target gene CHD5 to human colorectal cancer LOVO cell, and CHD5 can inhibited the proliferation of colorectal cancer LOVO cell.

Key words: CHD5, lentiviral vector, tumor

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