Targeting Ku protein for sensitizing of breast cancer cells to DNA-damage.
Targeting molecular components that are critically involved in the maintenance of genome stability is a promising approach for overcoming intrinsic tumor cell resistance to DNA-damaging treatments. In mammalian cells, the Ku-dependent non-homologous end-joining repair pathway is the predominant process for the repair of double-strand breaks (DSBs) in DNA. Previously, RNA aptamers were selected to efficiently block DNA-binding activity of the Ku protein in vitro. In the present study, we have tested the efficacy of RNA aptamers against the Ku protein as molecular sensitizer of MCF-7 breast carcinoma cells to DNA-damage. Toward this end, we established MCF-7 cell sublines stably expressing SC4 aptamer RNAs under the control of the human 7SL small nuclear RNA gene promoter. Vector-transfected (MCF/7SL) cells and cells stably expressing SC4 aptamers (MCF/SC4) were exposed to the anticancer drug etoposide and cellular responses to DNA-damage were evaluated. We found that the presence of RNA aptamers against Ku protein enhanced etoposide-induced growth inhibition of MCF-7 breast cancer. The SC4 aptamer-mediated sensitization of MCF-7 cells to the anticancer drug is attributable to an increased susceptibility of these cells to apoptosis. The observed effects cannot be accounted for by the differential expression levels of Ku protein in control and SC4 aptamer-expressing cells, but are rather due to augmented DNA binding-capacity of Ku protein, as demonstrated in in vitro studies. Thus, RNA aptamers against Ku protein show potential to sensitize MCF-7 breast carcinoma cells to DNA-damaging agents.
Zhang
Li
L
Department of Radiation Medicine, Lombardi Cancer Center, Georgetown University, Medical Center, Washington, DC 20057, USA.
Yoo
Sunghan
S
Dritschilo
Anatoly
A
Belyaev
Igor
I
Soldatenkov
Viatcheslav
V
D034961:Antigens, Nuclear / Q000737:chemistry* / Q000378:metabolism; D017209:Apoptosis; D015152:Blotting, Northern; D001943:Breast Neoplasms / Q000188:drug therapy / Q000473:pathology* / Q000532:radiotherapy*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002469:Cell Separation; D004249:DNA Damage; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D016521:Electrophoresis, Gel, Pulsed-Field; D005434:Flow Cytometry; D006801:Humans; D015151:Immunoblotting; D007700:Kinetics; D000072200:Ku Autoantigen; D012313:RNA / Q000737:chemistry; D011838:Radiation-Sensitizing Agents / Q000627:therapeutic use*; D013997:Time Factors
D034961:Antigens, Nuclear; D004268:DNA-Binding Proteins; D011838:Radiation-Sensitizing Agents; D012313:RNA; C000606509:Xrcc6 protein, human; D000072200:Ku Autoantigen