Knockdown of prion protein (PrP) by RNA interference weakens the protective activity of wild-type PrP against copper ion and antagonizes the cytotoxicity of fCJD-associated PrP mutants in cultured cells

  • Authors:
    • Zhao-Yun Wang
    • Chan Tian
    • Yuan-Yuan Jing
    • Han-Shi Gong
    • Yan Guo
    • Qi Shi
    • Cao Chen
    • Shu-Ying Zhu
    • Xiao-Ping Dong
  • View Affiliations

  • Published online on: May 2, 2011     https://doi.org/10.3892/ijmm.2011.688
  • Pages: 413-421
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Abstract

Development of the pathogenesis of transmissible spongiform encephalopathies (TSEs) requires the presence of both the normal host prion protein (PrPC) and the abnormal pathological proteinase-K resistant isoform (PrPSc). Reduction of PrPC levels has been shown to extend survival time after prion infection. In this report, based on analysis of the known sequences of human PrP, we constructed two small interfering RNA (siRNA) duplexes targeting the segments of amino acids (aa) 108-114 (Ri2) and aa 171-177 (Ri3). Western blot analysis results revealed that these PrP-specific siRNAs could effectively knock down the levels of either endogenous PrP in human neuroblastoma SHSY-5Y cells or recombinant PrP transfected with the plasmid expressing the full-length human PrP in human embryonic kidney (HEK) 293T cells. Meanwhile, the two siRNAs also showed a significant effect on the reduction of the expression of the PrP-PG9 and PrP-PG12 familial Creutzfeldt-Jakob disease (CJD)-associated PrP mutants with four and seven extra octarepeats, in the cells transfected with the respective expression plasmids. MTT tests identified that knockdown of wild-type PrP by Ri2 and Ri3 did not change the cell growth capacities, but significantly decreased the cell resistances against the challenge of Cu2+. Co-expression of Ri2 and Ri3 partially antagonized the cytotoxicity caused by expressing PrP-PG9 and PrP-PG12 in the two cell lines. Moreover, the rescuing effectiveness of PrP siRNAs was time-related, with the more efficient antagonism of the cytotoxicity of fCJD-associated PrP mutants occurring at the early stages after transfection. The data shown here provide useful clues for seeking potential therapeutic tools for prion diseases.

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September 2011
Volume 28 Issue 3

Print ISSN: 1107-3756
Online ISSN:1791-244X

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Spandidos Publications style
Wang Z, Tian C, Jing Y, Gong H, Guo Y, Shi Q, Chen C, Zhu S and Dong X: Knockdown of prion protein (PrP) by RNA interference weakens the protective activity of wild-type PrP against copper ion and antagonizes the cytotoxicity of fCJD-associated PrP mutants in cultured cells. Int J Mol Med 28: 413-421, 2011
APA
Wang, Z., Tian, C., Jing, Y., Gong, H., Guo, Y., Shi, Q. ... Dong, X. (2011). Knockdown of prion protein (PrP) by RNA interference weakens the protective activity of wild-type PrP against copper ion and antagonizes the cytotoxicity of fCJD-associated PrP mutants in cultured cells. International Journal of Molecular Medicine, 28, 413-421. https://doi.org/10.3892/ijmm.2011.688
MLA
Wang, Z., Tian, C., Jing, Y., Gong, H., Guo, Y., Shi, Q., Chen, C., Zhu, S., Dong, X."Knockdown of prion protein (PrP) by RNA interference weakens the protective activity of wild-type PrP against copper ion and antagonizes the cytotoxicity of fCJD-associated PrP mutants in cultured cells". International Journal of Molecular Medicine 28.3 (2011): 413-421.
Chicago
Wang, Z., Tian, C., Jing, Y., Gong, H., Guo, Y., Shi, Q., Chen, C., Zhu, S., Dong, X."Knockdown of prion protein (PrP) by RNA interference weakens the protective activity of wild-type PrP against copper ion and antagonizes the cytotoxicity of fCJD-associated PrP mutants in cultured cells". International Journal of Molecular Medicine 28, no. 3 (2011): 413-421. https://doi.org/10.3892/ijmm.2011.688