Construction of a Differentiated Embryo Chondrocyte 1 Lentiviral Expression Vector and Establishment of its Stably Transfected HGC27 Cell Line

  • Rui Hu
  • Yun-Shan Wang
  • Yi Kong
  • Pin Li
  • Yan Zheng
  • Xiao-Li Ma
  • Yan-Fei Jia
Conference paper
Part of the Lecture Notes in Electrical Engineering book series (LNEE, volume 269)

Abstract

Human differentiated embryo chondrocyte 1 (DEC1), has been suggested to play key roles in immune regulation, cell differentiation, proliferation and apoptosis, circadian rhythms, hypoxia response and carcinogenesis. However, the role of DEC1 in gastric cancer have not been well established. Lentiviral vectors are widely used for the stable expression of genes and currently under development for clinical use in gene therapy. Therefore we intended to construct a lentiviral DEC1 expression vector and then establish a gastric cancer cell line with stable expression of DEC1. The coding sequence of gene was amplified using PCR and cloned into pGV218 vector. 293T cells were transfected using Lipofectamine 2000 and packaged for the recombinant lentivirus particles. When the cloned sequence was identified to be right, the recombinant lentivirus particles were amplified in a large quantity. The titer of virus was determined by real-time PCR. Subsequently, we collected the lentivirus venom to infect the HGC27 cells and establish a stable, overexpressed cell line named GFP/DEC1-HGC27. This study will provide a new cell model for further study of the role of DEC 1in the pathogenesis of gastric cancer.

Keywords

DEC1 Lentiviral expression vector Gastric cancer HGC27 

Notes

Acknowledgments

This work was financially supported by National Natural Science Foundation of China (NSFC No. 81000869 and NSFC No. 81272588), 973 Project grant (2012CB966503 and 2012CB966504).

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Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Rui Hu
    • 1
  • Yun-Shan Wang
    • 1
    • 2
  • Yi Kong
    • 1
  • Pin Li
    • 1
  • Yan Zheng
    • 2
  • Xiao-Li Ma
    • 2
  • Yan-Fei Jia
    • 2
  1. 1.Shandong University School of MedicineJinanChina
  2. 2.Central Laboratory, Jinan Central HospitalAffiliated to Shandong UniversityJinanChina

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