A CD13 inhibitor, ubenimex, synergistically enhances the effects of anticancer drugs in hepatocellular carcinoma

Yamashita,Masafumi; Wada,Hiroshi; Eguchi,Hidetoshi; Ogawa,Hisataka; Yamada,Daisaku; Noda,Takehiro; Asaoka,Tadafumi; Kawamoto,Koichi; Gotoh,Kunihito; Umeshita,Koji; Doki,Yuichiro; Mori,Masaki

doi:10.3892/ijo.2016.3496

A CD13 inhibitor, ubenimex, synergistically enhances the effects of anticancer drugs in hepatocellular carcinoma

Authors:
- Masafumi Yamashita
- Hiroshi Wada
- Hidetoshi Eguchi
- Hisataka Ogawa
- Daisaku Yamada
- Takehiro Noda
- Tadafumi Asaoka
- Koichi Kawamoto
- Kunihito Gotoh
- Koji Umeshita
- Yuichiro Doki
- Masaki Mori
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Affiliations: Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan, Division of Health Sciences, Graduate School of Medicine, Osaka University, Osaka 565-0871, Japan
Published online on: April 25, 2016 https://doi.org/10.3892/ijo.2016.3496
Pages: 89-98
Copyright: © Yamashita et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Cancer stem cells (CSCs) were reported to be involved in resistance to chemo/radiation therapy. We previously reported that CD13 was both a marker of CSCs and a candidate therapeutic target in HCC. In the present study, we explored the antitumor effect of a combined therapy, where ubenimex, a CD13 inhibitor, was combined with conventional anticancer drugs, fluorouracil (5-FU), cisplatin (CDDP), doxorubicin (DXR) and sorafenib (SOR), and we elucidated the mechanism of these combination therapies. We evaluated changes in the expression of CD13 before and after treatment with anticancer drugs and with or without ubenimex in the human HCC cell lines HuH7 and PLC/PRF/5. The interactions between the anticancer drugs and ubenimex were determined with isobologram analyses. We analyzed cell cycle, apoptosis, and intracellular reactive oxygen species (ROS) levels to explore the mechanisms of the combination therapies. In both cell lines, the expression of CD13 increased after a 72-h exposure to each anticancer drug alone (p<0.05), and the expression of CD13 decreased with ubenimex administration (p<0.05). Isobologram analyses indicated that ubenimex had synergistic effects with 5-FU, CDDP and DXR, and an additive effect with SOR. Cell cycle analyses showed that ubenimex decreased the proportion of cells in G0/G1. Ubenimex enhanced the effects of 5-FU, CDDP and DXR by increasing apoptosis and intracellular ROS levels. In combination therapies, ubenimex synergistically enhanced the antitumor effects of 5-FU, CDDP and DXR on cell cycle regulation and apoptosis induction in HCC cell lines. The effects of ubenimex were due to increased intracellular ROS levels.

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