Aspirin inhibits hepatocellular carcinoma cell proliferation in vitro and in vivo via inducing cell cycle arrest and apoptosis

Shi,Tingting; Fujita,Koji; Gong,Jian; Nakahara,Mai; Iwama,Hisakazu; Liu,Shi; Yoneyama,Hirohito; Morishita,Asahiro; Nomura,Takako; Tani,Joji; Takuma,Kei; Tadokoro,Tomoko; Himoto,Takashi; Oura,Kyoko; Tsutsui,Kunihiko; Kobara,Hideki; Masaki,Tsutomu

doi:10.3892/or.2020.7630

Aspirin inhibits hepatocellular carcinoma cell proliferation in vitro and in vivo via inducing cell cycle arrest and apoptosis

Authors:
- Tingting Shi
- Koji Fujita
- Jian Gong
- Mai Nakahara
- Hisakazu Iwama
- Shi Liu
- Hirohito Yoneyama
- Asahiro Morishita
- Takako Nomura
- Joji Tani
- Kei Takuma
- Tomoko Tadokoro
- Takashi Himoto
- Kyoko Oura
- Kunihiko Tsutsui
- Hideki Kobara
- Tsutomu Masaki
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Affiliations: Department of Gastroenterology and Neurology, Faculty of Medicine, Kagawa University, Kida, Kagawa 761‑0793, Japan, Department of Gastroenterology, The First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116011, P.R. China, Life Science Research Center, Kagawa University, Kida, Kagawa 761‑0793, Japan
Published online on: May 29, 2020 https://doi.org/10.3892/or.2020.7630
Pages: 457-468
Copyright: © Shi et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Aspirin, a nonsteroidal anti‑inflammatory drug (NSAID), is known to inhibit cell proliferation in a variety of cancers. However, the underlying mechanism of this inhibition remains unknown. We investigated the effects of aspirin on hepatocellular carcinoma (HCC) cells using in vitro and in vivo models. Six HCC cell lines and a liver cancer cell line including Huh‑7 were used in assays that evaluated cell proliferation, cell cycle, and apoptosis. Flow cytometry, enzyme‑linked immunosorbent assay (ELISA), western blot analysis, and phosphorylated receptor tyrosine kinase array were used to evaluate the effects of aspirin on the cells, and microRNAs (miRNAs) were analyzed by a miRNA array chip. The results were validated in vivo using a nude mouse model of Huh‑7‑xenografted tumors. Our results showed that aspirin exhibited an antiproliferative effect on all cell lines. Moreover, aspirin induced G0/G1 cell cycle arrest and modulated the levels of cell cycle‑related molecules such as cyclin E, cyclin D1, and cyclin‑dependent kinase 2 (Cdk2). In addition, aspirin upregulated the levels of caspase‑cleaved cytokeratin 18, increased the proportion of early apoptotic cells, decreased the levels of clusterin and heat shock protein 70 (HSP 70), upregulated the levels of miRNA‑137 and inhibited epidermal growth factor receptor (EGFR) activation. In addition, we observed that aspirin suppressed cell proliferation partially through the miRNA‑137/EGFR pathway. Our in vivo results showed that aspirin reduced the growth of xenograft tumors in nude mice. In conclusion, aspirin was able to inhibit the growth of HCC cells by cell cycle arrest, apoptosis, and alteration of miRNA levels in in vitro and in vivo models.

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