Autophagy inhibition augments resveratrol‑induced apoptosis in Ishikawa endometrial cancer cells

Fukuda,Tomohiko; Oda,Katsutoshi; Wada‑Hiraike,Osamu; Sone,Kenbun; Inaba,Kanako; Ikeda,Yuji; Makii,Chinami; Miyasaka,Aki; Kashiyama,Tomoko; Tanikawa,Michihiro; Arimoto,Takahide; Yano,Tetsu; Kawana,Kei; Osuga,Yutaka; Fujii,Tomoyuki

doi:10.3892/ol.2016.4978

Autophagy inhibition augments resveratrol‑induced apoptosis in Ishikawa endometrial cancer cells

Authors:
- Tomohiko Fukuda
- Katsutoshi Oda
- Osamu Wada‑Hiraike
- Kenbun Sone
- Kanako Inaba
- Yuji Ikeda
- Chinami Makii
- Aki Miyasaka
- Tomoko Kashiyama
- Michihiro Tanikawa
- Takahide Arimoto
- Tetsu Yano
- Kei Kawana
- Yutaka Osuga
- Tomoyuki Fujii
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Affiliations: Department of Obstetrics and Gynecology, Graduate School of Medicine, The University of Tokyo, Tokyo 113‑8655, Japan, Department of Obstetrics and Gynecology, National Center for Global Health and Medicine, Tokyo 162‑0052, Japan
Published online on: August 8, 2016 https://doi.org/10.3892/ol.2016.4978
Pages: 2560-2566
Copyright: © Fukuda et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Resveratrol (RSV), a polyphenolic compound derived from red wine, inhibits the proliferation of various types of cancer. RSV induces apoptosis in cancer cells, while enhancing autophagy. Autophagy promotes cancer cell growth by driving cellular metabolism, which may counteract the effect of RSV. The present study aimed to elucidate the correlation between RSV and autophagy and to examine whether autophagy inhibition may enhance the antitumor effect of RSV in endometrial cancer cells. Cell proliferation, cell cycle progression and apoptosis were examined, following RSV exposure, by performing MTT assays, flow cytometry and annexin V staining, respectively, in an Ishikawa endometrial cancer cell line. Autophagy was evaluated by measuring the expression levels of light chain 3, II (LC3‑II; an autophagy marker) by western blotting and immunofluorescence. Chloroquine (CQ) and small interfering RNAs targeting autophagy related (ATG) gene 5 (ATG5) or 7 (ATG7) were used to inhibit autophagy, and the effects in combination with RSV were assessed using MTT assays. RSV treatment suppressed cell proliferation in a dose‑dependent manner in Ishikawa cells. In addition, RSV exposure increased the abundance of the sub‑G1 population and induced apoptosis. LC3‑II accumulation was observed following RSV treatment, indicating that RSV induced autophagy. Combination treatment with CQ and RSV more robustly suppressed growth inhibition and apoptosis, compared with RSV treatment alone. Knocking down ATG5 or ATG7 expression significantly augmented RSV‑induced apoptosis. The results of the present study indicated that RSV‑induced autophagy may counteract the antitumor effect of RSV in Ishikawa cells. Combination treatment with RSV and an autophagy inhibitor, such as CQ, may be an attractive therapeutic option for treating certain endometrial cancer cells.

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