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.
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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

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Figure 9

Petasin inhibits cellular motility and invasion of tumor cells.

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Petasin inhibits cellular motility and invasion of tumor cells. (A) Time...
(A) Time-course wound closure percentage and representative images in the scratch wound healing assay for tumor (B16F10) and nontumor (ASF 4-1) cells treated with different concentrations of petasin (PT; 0, 15, or 150 nM). White dotted lines, wound borders at 0 hours. Scale bar: 200 μm. (B) ATP/ADP ratio of B16F10 cells treated for 24 or 48 hours with PT (150 nM). Note that cell migration was already inhibited within 24 hours before the drop in the ATP/ADP ratio. (C) Experimental protocol for the Matrigel invasion assay. (D) Representative images and counts for cells that invaded the Matrigel and passed through the PET membrane in C (arrow, cells that invaded; arrowhead, pores of the PET membrane). (E) Experimental protocol for the cell attachment assay. (F) Representative images, counts for cells that attached to the surface of the culture plate within 3 hours (arrow, attached cells; arrowhead, nonattached cells; scale bar: 100 μm), and the ATP/ADP ratio in E. (G and H) Counts of focal adhesion spots (G) and representative confocal microscopic images of immunofluorescence staining for p-FAKY397 (yellow in the merged image), F-actin (phalloidin, red), and nuclei (DAPI, blue, H) in B16F10 cells treated with PT (0.3 μM) or DMSO (scale bar: 20 μm). (I) Immunoblot for focal adhesion-associated markers in tumor and nontumor cells treated with PT (3 μM) or DMSO (loading control, β-actin). (J) Pulldown assay for detecting the active form of Rac (Rac-GTP) in B16F10 cells treated for 48 hours with PT (0.3 μM) or DMSO in DMEM supplemented with 10% FBS. Data are presented as the mean ± SD (n = 3). *P < 0.05, **P < 0.01, ****P < 0.0001; 2-tailed, unpaired Student’s t test. NS, not significant High-glucose DMEM was used for the assays unless otherwise indicated.