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Arctigenin suppresses cell proliferation via autophagy inhibition in hepatocellular carcinoma cells

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Abstract

Autophagy is a catabolic process that degrades dysfunctional proteins and organelles and plays critical roles in cancer development. Our preliminary screening identified that extracts of the fruits of Arctium lappa and the fruits of Forsythia suspensa notably suppressed the proliferation of hepatocellular carcinoma HepG2 cells and downregulated the autophagy. In this study, we explored the effect of arctigenin (ARG), a bioactive lignan in both extracts, on cell proliferation and autophagy-related proteins in HepG2 cells. ARG inhibited the proliferation of HepG2 cells. Analysis of autophagy-related proteins demonstrated that ARG might block the autophagy that leads to sequestosome 1/p62 (p62) accumulation. The stage of inhibition in autophagy by ARG differed from those by the autophagy inhibitors 3-methyladenine (3-MA) or chloroquine (CQ). ARG could also inhibit starvation-induced autophagy. Further analysis of apoptosis-related proteins indicated that ARG did not affect caspase-3 activation and PARP cleavage, suggesting that the antiproliferative effect of ARG can occur independently of apoptosis. In summary, our study showed that ARG suppresses cell proliferation and inhibits autophagy, and might lead to the development of agents for autophagy research and cancer chemoprevention.

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Abbreviations

3-MA:

3-Methyladenine

ARC:

Arctiin

ARG:

Arctigenin

CQ:

Chloroquine

DAPI:

4′,6-Diamidino-2-phenylindole

DMEM:

Dulbecco’s modified Eagle medium

DMSO:

Dimethyl sulfoxide

ETP:

Etoposide

FBS:

Fetal bovine serum

HCC:

Hepatocellular cancer

LC3:

Light chain 3

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetra-zolium bromide

p62:

Sequestosome 1/p62

PBS:

Phosphate-buffered saline

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Acknowledgements

This study was partly supported by the Sasakawa Scientific Research Grant from the Japan Science Society (Grant no. 2018-4039).

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Authors and Affiliations

  1. Graduate School of Pharmaceutical Sciences, Nagasaki International University, 2825-7 Huis Ten Bosch-Cho, Sasebo, Nagasaki, 859-3298, Japan

    Shinya Okubo & Takuhiro Uto

  2. Faculty of Pharmaceutical Sciences, Nagasaki International University, 2825-7 Huis Ten Bosch-Cho, Sasebo, Nagasaki, 859-3298, Japan

    Tomoe Ohta, Yukihiro Shoyama & Takuhiro Uto

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  1. Shinya Okubo
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  2. Tomoe Ohta
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Correspondence to Takuhiro Uto.

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Okubo, S., Ohta, T., Shoyama, Y. et al. Arctigenin suppresses cell proliferation via autophagy inhibition in hepatocellular carcinoma cells. J Nat Med 74, 525–532 (2020). https://doi.org/10.1007/s11418-020-01396-8

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