Factor-inhibiting HIF (FIH) promotes lung cancer progression
Ana García-del Río, Endika Prieto-Fernández, Leire Egia-Mendikute, Asier Antoñana-Vildosola, Borja Jimenez-Lasheras, So Young Lee, Adrián Barreira-Manrique, Samanta Romina Zanetti, Ander de Blas, Paloma Velasco-Beltrán, Alexandre Bosch, Ana M. Aransay, Asis Palazon
Ana García-del Río, Endika Prieto-Fernández, Leire Egia-Mendikute, Asier Antoñana-Vildosola, Borja Jimenez-Lasheras, So Young Lee, Adrián Barreira-Manrique, Samanta Romina Zanetti, Ander de Blas, Paloma Velasco-Beltrán, Alexandre Bosch, Ana M. Aransay, Asis Palazon
View: Text | PDF
Research Article Cell biology

Factor-inhibiting HIF (FIH) promotes lung cancer progression

Abstract

Factor-inhibiting HIF (FIH) is an asparagine hydroxylase that acts on hypoxia-inducible factors (HIFs) to control cellular adaptation to hypoxia. FIH is expressed in several tumor types, but its impact in tumor progression remains largely unexplored. We observed that FIH was expressed on human lung cancer tissue. Deletion of FIH in mouse and human lung cancer cells resulted in an increased glycolytic metabolism, consistent with increased HIF activity. FIH-deficient lung cancer cells exhibited decreased proliferation. Analysis of RNA-Seq data confirmed changes in the cell cycle and survival and revealed molecular pathways that were dysregulated in the absence of FIH, including the upregulation of angiomotin (Amot), a key component of the Hippo tumor suppressor pathway. We show that FIH-deficient tumors were characterized by higher immune infiltration of NK and T cells compared with FIH competent tumor cells. In vivo studies demonstrate that FIH deletion resulted in reduced tumor growth and metastatic capacity. Moreover, high FIH expression correlated with poor overall survival in non–small cell lung cancer (NSCLC). Our data unravel FIH as a therapeutic target for the treatment of lung cancer.

Authors

Ana García-del Río, Endika Prieto-Fernández, Leire Egia-Mendikute, Asier Antoñana-Vildosola, Borja Jimenez-Lasheras, So Young Lee, Adrián Barreira-Manrique, Samanta Romina Zanetti, Ander de Blas, Paloma Velasco-Beltrán, Alexandre Bosch, Ana M. Aransay, Asis Palazon

×

Figure 1

FIH is expressed in lung cancer tissue and regulates HIF-driven metabolism.

Options: View larger image (or click on image) Download as PowerPoint
FIH is expressed in lung cancer tissue and regulates HIF-driven metaboli...
(A) Representative IHC images showing FIH expression (peroxidase-DAB brown staining) in normal tissue and lung cancer. Enlarged images are shown in the insets. Scale bars: 500 μm. (B) Western blot showing FIH expression in the indicated WT or FIH-KO cell lines generated by CRISPR/Cas9. (C) Western blot showing FIH expression in the indicated WT or FIH-knockdown human cell lines generated by short hairpin RNA (shRNA). (D) HRE luciferase reporter activity assay performed with LLC cells cultured in the presence or absence of 1 mM DMOG under the indicated conditions (n = 3, technical replicates are shown, unpaired t test). (E) Relative expression of the indicated HIF target genes in WT or FIH-KO LLC cells (n = 3, unpaired t test). (F) Immunoblotting showing HIF1A, PHD2, VHL, and tubulin expression in WT or FIH-KO LLC cells. (G) Flow cytometry analysis of glucose uptake (2-NBDG) in WT (black) or FIH-KO (red) LLC cells cultured under normoxia or hypoxia for 48 hours. A representative histogram indicating the geometric mean fluorescent intensity (gMFI) value for each condition (unstained, gray; WT, black or FIH-KO, red) is shown (n = 3, 2-way ANOVA). (H) Cell glucose consumption in WT or FIH-KO LLC cells cultured under normoxia or hypoxia for 48 hours measured by a colorimetric enzymatic assay (n = 3, unpaired t test). (I) Lactate production in WT or FIH-KO LLC cells cultured under normoxia or hypoxia for 48 hours and 72 hours measured by a colorimetric enzymatic assay (n = 3, unpaired t test). Cells were cultured for 16 hours under normoxia or hypoxia in B–F. Data are shown as mean ± SEM. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, and ****P ≤ 0.0001.