Journal: Cancer cell
Article Title: Leveraging an NQO1 Bioactivatable Drug for Tumor-Selective Use of Poly(ADP-ribose) Polymerase (PARP) Inhibitors
Figure Lengend Snippet: PARP inhibition and β-lapachone synergy is NQO1-dependent and broadly applied to various types of NQO1 over-expressing cancers (A,B) Polymorphic *2 H596 NSCLC cells corrected for NQO1 expression (A) were pretreated with Rucaparib (15 µM, 2 hr) and then exposed or not to Rucaparib (15 µM) + β-lap (0–4 µM) for 2 hr. Cells were also exposed to β-lap, ± dicoumarol (DIC, 50 µM) for 2 hr and survival assessed. Genetically matched NQO1-deficient H596 NSCLC cells (B) were treated as in (A) , and survival assessed. (C,D) NQO1 + MiaPaCa2 PDA cells (C) were pretreated with Rucaparib (15 µM, 2 hr), then exposed or not to Rucaparib (15 µM) + β-lap (0–3 µM or 0–6 µM for D , respectively) for 2 hr, ± dicoumarol (DIC, 50 µM). Drugs were removed and survival assessed. Stable shRNA- NQO1 down MiaPaCa2 (clone 17–7) vs shSCR MiaPaCa2 cells (D) were treated as in (C) , but without dicoumarol and assessed for survival. (E,F) NQO1 + Suit2 (S2-013) PDA cells (E) harboring a CMV-NQO1 over-expression vector (see Western, inset) were pretreated with Rucaparib (15 µM) and then exposed or not to β-lap (0–4 µM), ± dicoumarol (DIC, 50 µM) for 2 hr. Genetically matched, NQO1 − *2 polymorphic S2-013 chemo- and radio-resistant PDA cells (F) expressing shSCR were treated as in (E) and survival assessed. See inset (E) for NQO1 expression. (G,H) MDA-MB-231 *2 polymorphic TNBC cells corrected for NQO1 expression (G) were pretreated with Rucaparib (15 µM, 2 hr), + β-lap (0–2.5 µM), ± dicoumarol (DIC, 50 µM) for 2 hr. Drugs were removed and survival assessed. shSCR NQO1 − MDA-MB-231 TNBC cells (H) lacking NQO1 expression were treated as in (G) and assessed for survival. ( A–H ). Synergy values (η=0.452, p value=0.0003) were reported based on multiple dose-responses, or on comparative p values indicated. All error bars are means of six replicates from three independent experiments; means ± SEM. ***, p
Article Snippet: We show that the NAD(P)H:quinone oxidoreductase 1 (NQO1) bioactivatable drug, β-lapachone (ARQ761, ArQule, Woburn, MA, in clinical form), capitalizes on elevated NQO1:CAT ratios in recalcitrant pancreatic, non-small cell lung (NSCLC) and breast cancers to elicit tumor-selective programmed necrosis.
Techniques: Inhibition, Expressing, shRNA, Over Expression, Plasmid Preparation, Western Blot, Multiple Displacement Amplification