Knockdown strategies for the study of proprotein convertases and proliferation in prostate cancer cells.

Abstract

Gene silencing strategies targeting mRNA are suitable methods to validate the functions of specific genes. In this chapter, we sought to compare two knockdown strategies for the study of proprotein convertases and proliferation in prostate cancer cells. We used both SOFA-HDV ribozyme and lentiviral-mediated shRNA delivery system to reduce PACE4 mRNA levels and validate its implication in the proliferation of DU145 prostate cancer cells. The cellular effects of PACE4 knockdown were assessed (1) in vitro using two tetrazolium salts (MTT and XTT assays) and (2) in vivo using a tumor xenograft approach in immunodeficient mice (Nu/Nu). Our results confirm the unique role of the proprotein convertase PACE4 in prostate cancer cell proliferation while demonstrating advantages and disadvantages of each approach. Achieving target validation in an effective manner is critical, as further development using a drug development approach is highly laborious and requires enormous resources.

• Authors: François D'Anjou, Frédéric Couture, Roxane Desjardins, Robert Day
• Journal: Methods in molecular biology (Clifton, N.J.) (Methods Mol Biol) Vol. 1103 Pg. 67-82 ( 2014) ISSN: 1940-6029 [Electronic] United States
• PMID: 24318887 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• RNA, Catalytic
• RNA, Messenger
• RNA, Small Interfering
• PCSK6 protein, human
• Proprotein Convertases
• Serine Endopeptidases

Topics

• Animals
• Cell Line, Tumor
• Cell Proliferation
• Gene Knockdown Techniques
• Gene Silencing
• Humans
• Male
• Mice
• Molecular Biology (methods)
• Proprotein Convertases (genetics, metabolism)
• Prostatic Neoplasms (genetics, metabolism, pathology)
• RNA, Catalytic (genetics)
• RNA, Messenger (genetics)
• RNA, Small Interfering (genetics)
• Serine Endopeptidases (genetics, metabolism)