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Degradation of HIF-1α induced by curcumol blocks glutaminolysis and inhibits epithelial-mesenchymal transition and invasion in colorectal cancer cells

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Abstract

Colorectal cancer (CRC) has high morbidity and mortality. Epithelial-mesenchymal transition (EMT) is associated with CRC progression and metastasis. Glutaminolysis is essential for malignancy of cancer cells. Here, we examined the effects of curcumol on CRC EMT. We observed that curcumol suppressed invasion and migration in human CRC cells associated with upregulation of epithelial markers E-cadherin and Zonula occludens 1 and downregulation of mesenchymal markers N-cadherin and Vimentin as well as EMT-related transcription factors Snail and Twist. Curcumol increased intracellular levels of glutamine but decreased intracellular levels of glutamate, α-ketoglutarate, ATP, glutathione, and tricarboxylic acid cycle metabolites, suggesting interruption of glutaminolysis. Next, curcumol repressed glutaminase 1 (Gls1) mRNA and protein expression, and overexpression of Gls1 promoted EMT and abolished curcumol effects on CRC cell EMT. Molecular examinations showed that curcumol stimulated protein degradation of hypoxia-inducible factor-1α (HIF-1α) and prevented its nuclear accumulation in CRC cells. HIF-1α agonist deferoxamine (DFO) promoted HIF-1α binding to Gls1 promoter and increased Gls1 expression but abolished curcumol’s inhibitory effects on Gls1 expression. DFO also enhanced EMT and invasion and migration in CRC cells and eliminated curcumol effects. Furthermore, mouse CRC models were established with in vivo overexpression of HIF-1α and Gls1. Curcumol effectively inhibited CRC growth, metastasis, and EMT in mice, which was abrogated by overexpression of HIF-1α or Gls1. Altogether, stimulation of HIF-1α degradation was required for curcumol to disrupt EMT and repress invasion and migration in CRC cells through inhibiting Gls1-mediated glutaminolysis. Curcumol could be a promising candidate for intervention of CRC metastasis.

Graphical abstract

• Curcumol inhibits EMT and blocks glutaminolysis in CRC cells.

• Inhibition of Gls1 is required for curcumol blockade of glutaminolysis and EMT.

• Curcumol induces HIF-1α degradation leading to inhibition of Gls1 and blockade of glutaminolysis and EMT.

• Curcumol suppresses CRC growth and metastasis via inhibiting HIF-1α, glutaminolysis and EMT in mice.

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Abbreviations

α-KG:

α-Ketoglutarate

ChIP:

Chromatin immunoprecipitation

CHX:

Cycloheximide

CRC:

Colorectal cancer

DFO:

Deferoxamine

DMSO:

Dimethylsulfoxide

EMSA:

Electrophoretic mobility shift assay

EMT:

Epithelial-mesenchymal transition

FBS:

Fetal bovine serum

GDH:

Glutamate dehydrogenase

Gls1:

Glutaminase 1

GSH:

Glutathione

GSSG:

L-glutathione oxidized

HE:

Hematoxylin-eosin

HIF-1α:

Hypoxia inducible factor-1α

PBS:

Phosphate buffered saline

ROS:

Reactive oxygen species

TCA:

Tricarboxylic acid

ZO-1:

Zonula occludens 1

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Availability of data and material

All the data in the current research are available from the corresponding author on reasonable request.

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Funding

This work was supported by the National Natural Science Foundation of China (81873021 and 82074035), the Innovative Project of Postgraduate Training of Jiangsu Province (KYCX20_1486), and the Major Project for Science and Technology Development of Traditional Chinese Medicine of Jiangsu Province (2020ZX01).

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Author notes

  1. Yong Bian and Gang Yin contributed equally to this work.

Authors and Affiliations

  1. Laboratory Animal Center, Nanjing University of Chinese Medicine, Nanjing, 210023, China

    Yong Bian & Gang Wang

  2. School of Traditional Chinese Medicine and School of Integrated Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing, 210023, China

    Gang Yin, Tiantian Liu, Li Liang, Xinyue Yang & Decai Tang

  3. Jiangsu Key Laboratory of Oral Diseases, Nanjing Medical University, Nanjing, 211166, China

    Wen Zhang

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  1. Yong Bian
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Contributions

Decai Tang contributed to the design of the research and provided financial support. Yong Bian performed the experiments and analyzed the data. Gang Yin helped perform the cellular experiments. Gang Wang helped perform the animal experiments. Tiantian Liu conducted some other experiments. Li Liang helped analyze the data. Wen Zhang and Xinyue Yang revised the manuscript.

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Correspondence to Decai Tang.

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This study was approved by the Institutional and Local Committee on the Care and Use of Animals of Nanjing University of Chinese Medicine. All animals received humane care according to the National Institutes of Health (USA) guidelines.

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Bian, Y., Yin, G., Wang, G. et al. Degradation of HIF-1α induced by curcumol blocks glutaminolysis and inhibits epithelial-mesenchymal transition and invasion in colorectal cancer cells. Cell Biol Toxicol 39, 1957–1978 (2023). https://doi.org/10.1007/s10565-021-09681-2

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