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Article

Silencing of the human SET gene in vitro with lentivirus-mediated RNA interference

  • Authors:
    • Yanfeng Liu
    • Pengcheng He
    • Mei Zhang
    • Lili Shi
    • Huachao Zhu
    • Yuan Wang
    • Jing Zhao
  • View Affiliations / Copyright

    Affiliations: Department of Hematology, The First Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an 710061, P.R. China
  • Pages: 843-847
    |
    Published online on: January 15, 2013
       https://doi.org/10.3892/mmr.2013.1275
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Abstract

In our previous study, SET was identified as one of the differentially expressed proteins that was associated with tetra-arsenic tetra-sulfide (As4S4)-induced NB4-R1 [retinoic acid-resistant acute promyelocytic leukemia (APL) cell line] apoptosis. However, the mechanism through which SET regulates pathways during this process remains unclear. The aim of this study was to construct lentivirus-mediated short hairpin RNA (shRNA) against SET and investigate the effect of SET on As4S4-induced retinoic acid-resistant APL cell apoptosis. In the present study, 4 different oligonucleotides targeting the human SET gene were synthesized and cloned into the eukaryotic expression plasmid pGCSIL-GFP. The recombinant vectors were introduced into NB4-R1 cells. The silencing efficiency was measured by real-time quantitative PCR (RT-qPCR) and western blotting. Our results showed that the 4 recombinant RNA interference (RNAi) vectors were constructed successfully. Fluorescence microscopy demonstrated that infection efficiency ranged from 70 to 90%. Infection with the 4 different RNAi vectors significantly knocked down the expression of SET by 52.8, 69.1, 48.9 and 90.3% at the mRNA level, and 92.5, 96.3, 91.7 and 98.4% at the protein level, respectively. We attempt to clarify the mechanism of As4S4 treatment on retinoic acid-resistant APL.
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Copy and paste a formatted citation
Spandidos Publications style
Liu Y, He P, Zhang M, Shi L, Zhu H, Wang Y and Zhao J: Silencing of the human SET gene in vitro with lentivirus-mediated RNA interference. Mol Med Rep 7: 843-847, 2013.
APA
Liu, Y., He, P., Zhang, M., Shi, L., Zhu, H., Wang, Y., & Zhao, J. (2013). Silencing of the human SET gene in vitro with lentivirus-mediated RNA interference. Molecular Medicine Reports, 7, 843-847. https://doi.org/10.3892/mmr.2013.1275
MLA
Liu, Y., He, P., Zhang, M., Shi, L., Zhu, H., Wang, Y., Zhao, J."Silencing of the human SET gene in vitro with lentivirus-mediated RNA interference". Molecular Medicine Reports 7.3 (2013): 843-847.
Chicago
Liu, Y., He, P., Zhang, M., Shi, L., Zhu, H., Wang, Y., Zhao, J."Silencing of the human SET gene in vitro with lentivirus-mediated RNA interference". Molecular Medicine Reports 7, no. 3 (2013): 843-847. https://doi.org/10.3892/mmr.2013.1275
Copy and paste a formatted citation
x
Spandidos Publications style
Liu Y, He P, Zhang M, Shi L, Zhu H, Wang Y and Zhao J: Silencing of the human SET gene in vitro with lentivirus-mediated RNA interference. Mol Med Rep 7: 843-847, 2013.
APA
Liu, Y., He, P., Zhang, M., Shi, L., Zhu, H., Wang, Y., & Zhao, J. (2013). Silencing of the human SET gene in vitro with lentivirus-mediated RNA interference. Molecular Medicine Reports, 7, 843-847. https://doi.org/10.3892/mmr.2013.1275
MLA
Liu, Y., He, P., Zhang, M., Shi, L., Zhu, H., Wang, Y., Zhao, J."Silencing of the human SET gene in vitro with lentivirus-mediated RNA interference". Molecular Medicine Reports 7.3 (2013): 843-847.
Chicago
Liu, Y., He, P., Zhang, M., Shi, L., Zhu, H., Wang, Y., Zhao, J."Silencing of the human SET gene in vitro with lentivirus-mediated RNA interference". Molecular Medicine Reports 7, no. 3 (2013): 843-847. https://doi.org/10.3892/mmr.2013.1275
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