Transcription factor NRF2 regulates miR-1 and miR-206 to drive tumorigenesis
Anju Singh, … , Geoffrey Girnun, Shyam Biswal
Anju Singh, … , Geoffrey Girnun, Shyam Biswal
Published June 10, 2013
Citation Information: J Clin Invest. 2013;123(7):2921-2934. https://doi.org/10.1172/JCI66353.
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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 3

miR-1 and miR-206 miRNA target G6PD, PGD, TKT, and GPD2 in cancer cells and non-neoplastic fibroblast cells.

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miR-1 and miR-206 miRNA target G6PD, PGD, TKT, and GPD2 in cancer cells ...
(A) Sequence alignment showing the relative position of the miRNA binding site in the 3′UTR of each gene. (B) Expression of miR-1 and miR-206 decreases luciferase reporter gene activity in A549 cells when linked to the targeted segment of the 3′UTR of G6PD, PGD, TKT, and GPD2. Nonspecific control miR (NS miR) is a nontargeting control miRNA mimic. (C) Relative mRNA levels of G6PD, TKT, PGD, and GPD2 in A549 cells during expression of miR-1 and miR-206 as compared with those in cells expressing control miRNA. (D) Immunoblot analysis and (E) densitometric quantification showing relative protein levels of G6PD, TKT, PGD, and GPD2 in A549 cells during expression of miR-1 and miR-206 or control miRNA. (F) Relative levels of G6PD, TKT, PGD, and GPD2 in H1437 cells during expression of miR-1 and miR-206 as compared with those in the cells expressing nontargeting NS miRNA. Cotransfection of corresponding AMO along with precursor miRNA significantly blocks the action of miR-1 and miR-206. *P < 0.05 relative to the NS miR group.