In vitro selected RNA aptamer recognizing @entity2041 induces ROS mediated apoptosis in the human XXXX cell line MCF 7.

@entity522 @entity0 @entity5 @entity2041 @entity2197 @entity12274
@entity2041 ( @entity2197 ) is an abundant natural @entity12274 with antioxidant properties. Under different conditions, it can play protective as well as pathogenic roles. The redox state of the cell has an important role in the induction of apoptosis. Elevated level of @entity2041 in @entity5 cells provides resistance to a number of chemotherapeutic drugs. Inhibition of @entity2041 synthesis sensitizes the cells for apoptosis and enhances the activity of chemotherapeutic drugs. We have selected @entity2197 -binding RNA aptamers by employing in vitro selection protocol SELEX. The Kd value of these aptamers with respect to @entity2197 were determined by surface plasmon resonance (SPR) analysis and isocratic affinity chromatography. Two aptamers GSHapt 8.17 (class-III) and @entity2197 apt 5.39 (class-IV) had Kd values of 4.18 and 4.89 x 10(-8) M, respectively and GSHapt class-I had a Kd value of 1.2 x 10(-6) M. CD spectra suggested conformational change in aptamers upon @entity2197 binding. Cultured @entity0 cells (MCF7) responded to expression of @entity2197 aptamers by accumulating ROS and undergoing morphological transition, nuclear condensation, and DNA fragmentation, with concurrent depletion of cellular @entity2197 and activation of @entity522 eventually leading to apoptosis. DTT and @entity522 inhibitor partially rescued aptamer induced apoptosis. These aptamers exhibit high specificity to @entity2197 over non specific competitor. The same aptamers did not induce apoptosis in 293T cells. The kinetic properties and pro-apoptotic effects suggest that @entity2041 -binding RNA aptamer could be developed into an effective anti- @entity5 chemotherapeutic agent.
