Bcl-2 targeting siRNA expressed by a T7 vector system inhibits human tumor cell growth in vitro.

Abstract

A T7 promoter driven siRNA expression vector system (Bcl-2/T7) that targets Bcl-2 mRNA in MCF-7 human cancer cells was designed in the present study. In the presence of prebound T7 RNA polymerase, successful expression of Bcl-2 siRNA as well as its function was demonstrated via cell proliferation assays, Bcl-2 Elisa, and TUNEL assay. MCF-7 breast cancer cells transfected with Bcl-2/T7 show decreased levels of Bcl-2 expression at the protein level as well as decreased cell proliferation. Also, the number of apoptotic cells was increased in cells expressing Bcl-2 siRNA. Previous studies have shown that Bcl-2 levels are increased in a large number of different types of cancer. Therefore, the ability of Bcl-2/T7 to produce functional Bcl-2 siRNA in breast cancer cells suggests a potential role for this delivery system in cancer gene therapy.

• Authors: Lori Holle, Labri Hicks, Wendy Song, Eric Holle, Thomas Wagner, Xianzhong Yu
• Journal: International journal of oncology (Int J Oncol) Vol. 24 Issue 3 Pg. 615-21 (Mar 2004) ISSN: 1019-6439 [Print] Greece
• PMID: 14767546 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Luminescent Proteins
• Proto-Oncogene Proteins c-bcl-2
• RNA, Small Interfering
• Viral Proteins
• Green Fluorescent Proteins
• bacteriophage T7 RNA polymerase
• DNA-Directed RNA Polymerases

Topics

• Apoptosis
• Base Sequence
• Breast Neoplasms (therapy)
• Cell Division
• Cell Line, Tumor
• Cell Separation
• DNA-Directed RNA Polymerases (genetics)
• Enzyme-Linked Immunosorbent Assay
• Flow Cytometry
• Genetic Therapy (methods)
• Green Fluorescent Proteins
• HeLa Cells
• Humans
• In Situ Nick-End Labeling
• Luminescent Proteins (metabolism)
• Molecular Sequence Data
• Plasmids (metabolism)
• Proto-Oncogene Proteins c-bcl-2 (biosynthesis, genetics)
• RNA, Small Interfering (metabolism)
• Time Factors
• Transcription, Genetic
• Viral Proteins