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Supplemental Figures 1-14 from Depleting Tumor-NQO1 Potentiates Anoikis and Inhibits Growth of NSCLC

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posted on 2023-04-03, 16:27 authored by Brian Madajewski, Michael A. Boatman, Gaurab Chakrabarti, David A. Boothman, Erik A. Bey

Supplemental Figure 1: Tumor-NQO1 overexpression leads to poor prognosis in lung cancer patients. Supplemental Figure 2: shNQO1-B knockdown of NQO1 leads to decreased growth in soft agar. Supplemental Figure 3: shNQO1-B knockdown of NQO1 in H292 cells leads to decreased growth in soft agar. Supplemental Figure 4: Dicoumarol inhibits growth of A549 cells in soft agar. Supplemental Figure 5: Mac220 inhibits growth of A549 cells in soft agar. Supplemental Figure 6: Stable overexpression of NQO1 in H596 (NQO1 null) lung cancer cells causes increased growth in soft agar. Supplemental Figure 7: Transient knockdown of NQO1 in H596 LPC-NQO1 cells causes decreased growth in soft agar. Supplemental Figure 8: shNQO1-B knockdown of NQO1 in A549 cells decreases cell invasion. Supplemental Figure 9: Stable overexpression of NQO1 in H596 NQO1 null cells causes increased invasion. Supplementary Figure 10: Transient knockdown of NQO1 using siRNA inhibits invasion of HCC1171 cells. Supplemental Figure 11: Stable knockdown of NQO1 in A549 cells causes loss of PARP- 1 protein expression. Supplemental Figure 12: Knockdown of NQO1 reduces ALDH (high) activity in Pancreas cancer cells. Supplemental Figure 13: Knockdown of NQO1 reduces ALDH (high) activity in Prostate cancer cells. Supplemental Figure 14: NQO1 knockdown does not inhibit short-term viability our long-term survival of non-transformed human bronchial epithelial cells.

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ARTICLE ABSTRACT

The fundamental role that NAD(P)H/quinone oxidoreductase 1 (NQO1) plays, in normal cells, as a cytoprotective enzyme guarding against stress induced by reactive oxygen species (ROS) is well documented. However, what is not known is whether the observed overexpression of NQO1 in neoplastic cells contributes to their survival. The current study discovered that depleting NQO1 expression in A549 and H292 lung adenocarcinoma cells caused an increase in ROS formation, inhibited anchorage-independent growth, increased anoikis sensitization, and decreased three-dimensional tumor spheroid invasion. These in vivo data further implicate tumor-NQO1 expression in a protumor survival role, because its depletion suppressed cell proliferation and decreased lung tumor xenograft growth. Finally, these data reveal an exploitable link between tumor-NQO1 expression and the survival of lung tumors because NQO1 depletion significantly decreased the percentage of ALDH(high) cancer cells within the tumor population.Implications: Loss of tumor-NQO1 expression inhibits tumor growth and suggests that novel therapeutics directed at tumor-NQO1 may have clinical benefit. Mol Cancer Res; 14(1); 14–25. ©2015 AACR.

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