Western blot, MDA, ROS, qPCR raw data for thesis ROLE AND REGULATION OF NUR77 IN HYPOXIC TUMOR MICROENVIRONMENT AND FERROPTOTIC TUMOR CELL DEATH.
Abstract: Hypoxia is a
common characteristic in the tumor microenvironment (TME) and a critical
stimulant of primary and metastatic colorectal cancer (CRC) growth and
progression. Orphan nuclear receptor Nur77 (NR4A1) is frequently overexpressed
in hypoxic CRC tissues and cell lines. In Chapter 3, Nur77 is shown to regulate
Dicer-mediated microRNA biogenesis that explain its pronounced tumor-promoting
activities in CRC cells under hypoxic conditions. Mechanistically, Nur77 repressed
Dicer, a pivotal microRNA processor, and subsequently reduced the biogenesis of
let-7i-5p, which targeted p110α mRNA 3’UTR and modulated p110α mRNA stability.
Nur77 depletion or let-7i-5p overexpression suppressed the CRC metastasis in
mouse model.
Furthermore, an
inhibitory role of a natural small molecule ginsenoside compound K (CK) in CRC
stemness (CSC) by targeting Nur77 in the hypoxic microenvironment is identified
in Chapter 4. CK potently inhibited CSCs self-renewal in the hypoxic CRC cells,
and that this was mediated through Nur77 via a p110α PIK3CA-Akt
signaling pathway. Importantly, CK could effectively reduce tumorigenesis and
metastasis in xenograft mice with no adverse effects on the liver, lungs,
spleen, heart, and kidneys in vivo. These findings suggest that CK could
be a nutraceutical with a beneficial effect against CRC through targeting CSCs.
Notably, a dose-dependent enhancement of ROS was found upon CK treatment. This
massive cellular oxidation and excessive lipid peroxidation by high levels of
ROS is known to link an iron-dependent mode of cell death ferroptosis. In
Chapter 5, a new role for CK in stimulating ferroptosis in cancer cell lines
under hypoxic conditions is studied. In search of the underlying mechanisms, Nur77
bound to the Gpx4 promoter region to inhibit its transcription. Nur77 was also
found to regulate SLC7A11 by protecting it from ubiquitination and proteasome
degradation. Taken together, these data uncovered a novel mechanistic role of
Nur77-PI3K/Akt signaling in CRC metastasis under hypoxia. CK could be exploited
further as a therapeutic strategy for CRC patients.