Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (50): 9433-9436.doi: 10.3969/j.issn.1673-8225.2011.50.030

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Construction and identification of endothelial cell lines stably expressing human heat shock protein 22

Bao Hui-hui, Chen Qi, Wang Ling, Cheng Xiao-shu   

  1. Department of Cardiology, Second Affiliated Hospital of Nanchang University, Institute of Cardiovascular Disease of Nanchang University, Nanchang  330006, Jiangxi Province, China
  • Received:2011-05-21 Revised:2011-06-26 Online:2011-12-10 Published:2011-12-10
  • Contact: Cheng Xiao-shu, Doctor, Professor, Doctoral supervisor, Department of Cardiology, Second Affiliated Hospital of Nanchang University, Institute of Cardiovascular Disease of Nanchang University, Nanchang 330006, Jiangxi Province, China xiaoshumenfan@126.com
  • About author:Bao Hui-hui☆, Doctor, Attending physician, Lecturer, Department of Cardiology, Second Affiliated Hospital of Nanchang University, Institute of Cardiovascular Disease of Nanchang University, Nanchang 330006, Jiangxi Province, China huihui_bao@hotmail.com
  • Supported by:

    the National Natural Science Foundation for Distinguished Young Scholars, No. 30800467*; the Natural Science Foundation of Jiangxi Province, No. 0640089*

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Abstract:

BACKGROUND: Stress expression of heat shock protein 22 (HSP22) may protect endothelial cells against hypoxia/reoxygenation injury.
OBJECTIVE: To establish endothelial cell line stably expressing human HSP22 gene.
METHODS: The eukaryotic expression vector pEGFP-N1-HSP22 was identified by EcoRⅠ/kpnⅠdouble enzyme digestion. The recombinant plasmids were transfected into human umbilical vein endothelial cells (HUVECs) by lipofectamine. After screening culture by G418, a stably transfected cell line was established. The transcription and expression of the HSP22 gene were identified by fluorescence microscope, RT-PCR and western-blot assay.
RESULTS AND CONCLUSION: Agarose gel electrophoresis detection showed that HSP22 DNA segment was in accordance with the expected results after enzyme digestion. Green fluorescent protein expression was observed in each cell through the use of fluorescence microscope. HSP22 mRNA and protein expression were detected by RT-PCR and western blot analysis. The HUVEC monoclonal cells with a stable expression of HSP22 gene have been established successfully.

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