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CD59 silencing via retrovirus-mediated RNA interference enhanced complement-mediated cell damage in ovary cancer

Xue Xiang Shi1, Bei Zhang, Jin Lin Zang

  • 1Department of Immunology, Medical College of Qingdao University, Qingdao 266071, China.

Cellular & Molecular Immunology
|March 4, 2009
PubMed

Insights

Silencing CD59 (a complement inhibitor) in ovary cancer cells enhances complement-mediated damage and inhibits tumor growth. This suggests CD59 is a potential target for ovarian cancer gene therapy.

Area of Science:

  • Oncology
  • Immunology
  • Molecular Biology

Background:

  • CD59, a membrane complement regulatory protein (mCRP), inhibits complement-mediated cell lysis.
  • Over-expression of CD59 is observed in solid tumors, including ovarian cancer.

Purpose of the Study:

  • To construct a recombinant retrovirus encoding shRNA targeting human CD59.
  • To investigate the effect of decreased CD59 expression on ovarian cancer tumorigenesis using A2780 cells.

Main Methods:

  • Construction and identification of siCD59 and siCD59-C retroviruses via PCR, restriction enzyme analysis, and DNA sequencing.
  • Confirmation of retroviral infection in A2780 cells using Western blotting.
  • Assessment of cell viability, damage, caspase-3 activation, and apoptosis (Hoechst staining) after serum incubation.
  • Evaluation of tumor growth and CD59 protein expression in vivo using nude mouse xenografts.

Main Results:

  • Retroviral vectors siCD59 and siCD59-C were successfully constructed and verified.
  • siCD59 efficiently infected A2780 cells, leading to decreased cell viability and increased cell damage upon complement exposure.
  • CD59 silencing induced caspase-3 activation and apoptosis in ovarian cancer cells, and significantly reduced tumor weight in vivo.
  • Reduced CD59 protein expression was confirmed in tumor tissues from the siCD59 group.

Conclusions:

  • Retrovirus-mediated RNA interference (RNAi) targeting CD59 enhances complement-mediated cytotoxicity in ovarian cancer cells.
  • CD59 silencing inhibits ovarian cancer growth, indicating its potential as a therapeutic target for gene therapy.

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