Transcription factor NRF2 regulates miR-1 and miR-206 to drive tumorigenesis
Anju Singh, Christine Happel, Soumen K. Manna, George Acquaah-Mensah, Julian Carrerero, Sarvesh Kumar, Poonam Nasipuri, Kristopher W. Krausz, Nobunao Wakabayashi, Ruby Dewi, Laszlo G. Boros, Frank J. Gonzalez, Edward Gabrielson, Kwok K. Wong, Geoffrey Girnun, Shyam Biswal
Anju Singh, Christine Happel, Soumen K. Manna, George Acquaah-Mensah, Julian Carrerero, Sarvesh Kumar, Poonam Nasipuri, Kristopher W. Krausz, Nobunao Wakabayashi, Ruby Dewi, Laszlo G. Boros, Frank J. Gonzalez, Edward Gabrielson, Kwok K. Wong, Geoffrey Girnun, Shyam Biswal
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Research Article Oncology

Transcription factor NRF2 regulates miR-1 and miR-206 to drive tumorigenesis

Abstract

The mechanisms by which deregulated nuclear factor erythroid-2–related factor 2 (NRF2) and kelch-like ECH-associated protein 1 (KEAP1) signaling promote cellular proliferation and tumorigenesis are poorly understood. Using an integrated genomics and 13C-based targeted tracer fate association (TTFA) study, we found that NRF2 regulates miR-1 and miR-206 to direct carbon flux toward the pentose phosphate pathway (PPP) and the tricarboxylic acid (TCA) cycle, reprogramming glucose metabolism. Sustained activation of NRF2 signaling in cancer cells attenuated miR-1 and miR-206 expression, leading to enhanced expression of PPP genes. Conversely, overexpression of miR-1 and miR-206 decreased the expression of metabolic genes and dramatically impaired NADPH production, ribose synthesis, and in vivo tumor growth in mice. Loss of NRF2 decreased the expression of the redox-sensitive histone deacetylase, HDAC4, resulting in increased expression of miR-1 and miR-206, and not only inhibiting PPP expression and activity but functioning as a regulatory feedback loop that repressed HDAC4 expression. In primary tumor samples, the expression of miR-1 and miR-206 was inversely correlated with PPP gene expression, and increased expression of NRF2-dependent genes was associated with poor prognosis. Our results demonstrate that microRNA-dependent (miRNA-dependent) regulation of the PPP via NRF2 and HDAC4 represents a novel link between miRNA regulation, glucose metabolism, and ROS homeostasis in cancer cells.

Authors

Anju Singh, Christine Happel, Soumen K. Manna, George Acquaah-Mensah, Julian Carrerero, Sarvesh Kumar, Poonam Nasipuri, Kristopher W. Krausz, Nobunao Wakabayashi, Ruby Dewi, Laszlo G. Boros, Frank J. Gonzalez, Edward Gabrielson, Kwok K. Wong, Geoffrey Girnun, Shyam Biswal

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Figure 4

Gain of NRF2 function in lung cancer upregulates the expression of PPP genes by suppressing the expression of miR-1 and miR-206.

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Gain of NRF2 function in lung cancer upregulates the expression of PPP g...
(A and B) Increased expression of miR-1 in NRF2-deficient A549-Nrf2 shRNA and DU145-Nrf2 shRNA cells compared with that in the Luc shRNA control group. *P < 0.05 relative to Luc shRNA. (C and D) Increased expression of PPP genes in A549-Nrf2 shRNA and DU145-Nrf2 shRNA cells expressing anti–miR-1. Cells stably expressing empty vector were used as the baseline control to calculate the fold change. *P ≤ 0.01 relative to the cells expressing empty vector. (E) Increased expression of miR-1 decreases luciferase reporter gene activity associated with the 3′UTR of G6PD, PGD, TKT, and GPD2 in DU145-Nrf2 shRNA cells. The 3′UTR-containing reporter plasmids and pGLO empty vector were transfected into DU145 parent and Nrf2 shRNA cells, and the extracts were analyzed for luciferase activity. Firefly luciferase activity was normalized to Renilla luciferase activity. *P ≤ 0.01 relative to the parent cell line. (F) Increased expression of miR-206 in Nrf2–/– MEF cells and reduced expression of miR-206 in Keap1–/– MEF cells. *P ≤ 0.01 relative to WT cells.