Bcl-2 targeting siRNA expressed by a T7 vector system inhibits human tumor cell growth in vitro.
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.
Holle
Lori
L
Oncology Research Institute of the Greenville Hospital System, Greenville, SC 29605, USA.
Hicks
Labri
L
Song
Wendy
W
Holle
Eric
E
Wagner
Thomas
T
Yu
Xianzhong
X
D017209:Apoptosis; D001483:Base Sequence; D001943:Breast Neoplasms / Q000628:therapy*; D002455:Cell Division; D045744:Cell Line, Tumor; D002469:Cell Separation; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D015316:Genetic Therapy / Q000379:methods*; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D020287:In Situ Nick-End Labeling; D008164:Luminescent Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D010957:Plasmids / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis* / Q000235:genetics*; D034741:RNA, Small Interfering / Q000378:metabolism*; D013997:Time Factors; D014158:Transcription, Genetic; D014764:Viral Proteins
D008164:Luminescent Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D034741:RNA, Small Interfering; D014764:Viral Proteins; D049452:Green Fluorescent Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases