Integrative Gene Expression Profiling Reveals G6PD-Mediated Resistance to RNA-Directed Nucleoside Analogues in B-Cell Neoplasms
Figure 4
RNAi-mediated G6PD suppression sensitizes resistant cell lines to both 8-NH2-Ado and 8-Cl-Ado.
(A) Resistant cell lines were transduced with lentiviral vectors encoding control, non-targeted or G6PD-targeted shRNAs. G6PD protein expression was analyzed by immunoblotting. Representative blots from three independent experiments are shown. (B) Densitometric quantification of G6PD expression from data in part (A) is shown and normalized to control shRNA-transduced cells. (C) Granta 519 cells were treated for 17 hours with the indicated concentrations of 8-NH2-Ado three days after transduction (n = 3). (D) U266 cell lines stably expressing distinct shRNAs were treated for 17 hours with the indicated concentrations of 8-NH2-Ado (n = 5). (E) Granta 519 cells were treated for 48 hours with the indicated concentrations of 8-Cl-Ado two days after transduction (n = 5). (F) U266 cell lines stably expressing distinct shRNAs were treated for 48 hours with the indicated concentrations of 8-Cl-Ado (n = 5). In parts (C)–(F), cell death was assessed solely by DAPI staining and data represent the increase in cell death relative to untreated cells (normalized to 0%). Data in parts (B)–(F) are means ± SEM. * P<.05 ** P<.01 *** P<.005.