Acta Academiae Medicinae Sinica

Acta Academiae Medicinae Sinica

Acta Academiae Medicinae Sinica ›› 2014, Vol. 36 ›› Issue (4): 420-425.doi: 10.3881/j.issn.1000-503X.2014.04.013

• Original article • Previous Articles     Next Articles

Establishment and Functional Identification of A Stable Cell Line Overexpression Heme Oxygenase-1

YANG Gen-huan1,WU Wei2,LI Yan-chuan2,NI Leng1,WANG Zhan-qi1,SONG Xi-tao1,LIU Chang-wei1   

  1. 1Department of Vascular Surgery,PUMC Hospital,CAMS and PUMC,Beijing 100730,China; 2State Key Laboratory of Medical Molecular Biology,Institute of Basic Medical Sciences,CAMS and PUMC,Beijing 100005,China
  • Received:2014-03-07 Online:2014-08-31 Published:2014-08-31
  • Contact: LIU Chang-wei Tel:010-69152501,E-mail:liucw@vip.sina.com
  • Supported by:
    Supported by the China Postdoctoral Science Foundation (2009045513)

Abstract: Objective To establish a stable cell line overexpression heme oxygenase-1(HO-1) mediated by a modified lentivirus system and identify its function. Methods The HO-1 gene was amplified by polymerase chain reaction and cloned into the modified pLentiLox3.7 expression vectors. The recombinant plasmids were transfected into HEK293T cells and the HO-1 was detected by Western blot. The recombinant plasmids were transfected into HEK293T cells to produce the viruses,with the helping plasmids including plp1,plp2,and VSVG. HEK293T cells were infected by the viruses and the cells that can express HO-1 were identified by Western blot. The reactive oxygen species were detected in the HO-1-overexpression HEK293T cells and the normal cells after the adding of hydrogen peroxide. The same experiment was performed with human umbilical vein endothelial cells. Results The stable cell line that can overexpress HO-1 was established,which was verified by Western blot. The reactive oxygen species in the HO-1-overexpression HEK293T cells and human umbilical vein endothelial cells decreased obviously after exposure to hydrogen peroxide. Conclusions The lentivirus-carrying HO-1 was successfully packaged and the stable cell line overexpression HO-1 was established. HO-1 can play a protective role in the course of oxidative damage.

Key words: heme oxygenase-1, lentivirus, stable cell lines, oxidative damage

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