Petasin potently inhibits mitochondrial complex I–based metabolism that supports tumor growth and metastasis
Kazuki Heishima, … , Hiroshi Ueda, Yukihiro Akao
Kazuki Heishima, … , Hiroshi Ueda, Yukihiro Akao
Published September 1, 2021
Citation Information: J Clin Invest. 2021;131(17):e139933. https://doi.org/10.1172/JCI139933.
View: Text | PDF
Research Article Metabolism Oncology Article has an altmetric score of 59

Petasin potently inhibits mitochondrial complex I–based metabolism that supports tumor growth and metastasis

Abstract

Mitochondrial electron transport chain complex I (ETCC1) is the essential core of cancer metabolism, yet potent ETCC1 inhibitors capable of safely suppressing tumor growth and metastasis in vivo are limited. From a plant extract screening, we identified petasin (PT) as a highly potent ETCC1 inhibitor with a chemical structure distinct from conventional inhibitors. PT had at least 1700 times higher activity than that of metformin or phenformin and induced cytotoxicity against a broad spectrum of tumor types. PT administration also induced prominent growth inhibition in multiple syngeneic and xenograft mouse models in vivo. Despite its higher potency, it showed no apparent toxicity toward nontumor cells and normal organs. Also, treatment with PT attenuated cellular motility and focal adhesion in vitro as well as lung metastasis in vivo. Metabolome and proteome analyses revealed that PT severely depleted the level of aspartate, disrupted tumor-associated metabolism of nucleotide synthesis and glycosylation, and downregulated major oncoproteins associated with proliferation and metastasis. These findings indicate the promising potential of PT as a potent ETCC1 inhibitor to target the metabolic vulnerability of tumor cells.

Authors

Kazuki Heishima, Nobuhiko Sugito, Tomoyoshi Soga, Masashi Nishikawa, Yuko Ito, Ryo Honda, Yuki Kuranaga, Hiroki Sakai, Ryo Ito, Takayuki Nakagawa, Hiroshi Ueda, Yukihiro Akao

×

Figure 2

Petasin induces cell-cycle arrest and necrotic cell death with ATP depletion.

Options: View larger image (or click on image) Download as PowerPoint
Petasin induces cell-cycle arrest and necrotic cell death with ATP deple...
(A) Cell-count percentage in the G0/G1 (2N), S, and G2/M (4N) phases for tumor or nontumor cells treated with PT (3 μM) or DMSO. (B) Immunoblots for cell-cycle markers in melanoma (B16F10, A2058), pancreatic cancer (MiaPaCa2), chronic myeloid leukemia (K562), and nontumor (ASF 4-1 and HMEC) cell lines. (C) Phase-contrast microscopic images of B16F10 cells treated for 0, 24, 60, or 72 hours with PT (3 μM) or DMSO in high-glucose DMEM. Scale bars: 20 μm. (D) Fold-change (FC) in pH and in glucose and lactate concentrations in medium of B16F10 or ASF 4-1 cell cultures treated for 72 hours with PT (3 μM) or DMSO in high-glucose DMEM. (E) Transmission electron microscopic images of B16F10 cells treated for 24 or 60 hours with PT (3 μM) or DMSO. Arrow and arrowheads indicate plasma membrane and mitochondria, respectively. Scale bars: 1 μm. (F) Viable cell percentage of B16F10 cells treated for 48 hours with PT (3 μM) in EMEM supplemented with or without glucose (Glu, final concentration of 4.5 g/L), essential amino acids (EAA), and nonessential amino acids (NEAA). ****P < 0.0001, 1-way ANOVA with Tukey’s post hoc test. (G) Time course of FC in viable cell count and ATP/ADP ratio. B16F10 cells were treated with PT (3 μM) in DMEM containing different concentrations of glucose (DMEM-NG, no glucose, 0 g/L; DMEM-LG, low glucose, 1 g/L; DMEM-HG, high glucose, 4.5 g/L). (H) Viable cell percentage and representative images for B16F10 cells treated with PT (3 μM) or DMSO in DMEM-HG with or without medium refreshment every 24 hours. ****P < 0.0001, 1-way ANOVA with Tukey’s post hoc test. Scale bars: 50 μm. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; 2-tailed, unpaired Student’s t test unless otherwise indicated. Data are presented as the mean ± SD (n = 3). NS, not significant.