Bowman-Birk protease inhibitor from soybeans enhances cisplatin-induced cytotoxicity in human mesothelioma cells

Kashiwagi,Korehito; Virgona,Nantiga; Yamada,Jin; Sato,Ayami; Ota,Masako; Yazawa,Takuya; Yano,Tomohiro

doi:10.3892/etm.2011.270

Bowman-Birk protease inhibitor from soybeans enhances cisplatin-induced cytotoxicity in human mesothelioma cells

Authors:
- Korehito Kashiwagi
- Nantiga Virgona
- Jin Yamada
- Ayami Sato
- Masako Ota
- Takuya Yazawa
- Tomohiro Yano
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Affiliations: Faculty of Life Sciences, Toyo University, Oura, Gunma 374-0192, Japan , Project for Complementary Medicine, National Institute of Health and Nutrition, Shinjuku, Tokyo 162-8636, Japan, Department of Pathology, Yokohama City University, Graduate School of Medicine, Kanazawa, Yokohama 236-0004, Japan
Published online on: May 12, 2011 https://doi.org/10.3892/etm.2011.270
Pages: 719-724

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Abstract

Malignant mesothelioma (MM) is an aggressive cancer with no effective treatment options. Enforced expression of the gap junction (GJ) component connexin 43 (Cx43) increases the sensitivity of MM cells to cisplatin. Bowman-Birk protease inhibitor (BBI) induces the restoration of Cx43 in several types of tumor cells. In this study, we examined the capability of BBI to enhance the cytotoxic effect of cisplatin in MM cells via the induction of Cx43. Human MM H28 cells were used. Cell viability was evaluated by a WST-1 assay and proteasomal activity was determined by fluorometric analysis. Protein and mRNA levels were determined by immunoblot analysis and real-time RT-PCR, respectively. GJ function mediated by Cx43 was evaluated using the scrape-loading method. BBI effectively inhibited H28 cell growth in a dose-dependent manner (200-400 µg/ml). In parallel with the growth inhibition, Cx43 levels (mRNA and protein) and GJ function were elevated by BBI treatment. Knockdown of BBI-induced Cx43 by an antisense nucleotide treatment almost cancelled the growth inhibition. BBI enhanced cisplatin-induced cytotoxicity in H28 cells, and down-regulation of Cx43 by the antisense nucleotide treatment abrogated the enhancing effect of BBI. The induction of Cx43 by BBI contributed to Src inactivation and subsequent induction of Bax. Furthermore, an Src inhibitor (SU6656) also enhanced cisplatin-induced cytotoxicity in H28 cells. These results suggest that BBI improves the cytotoxic efficacy of cisplatin in H28 cells via the inhibition of Src signaling.

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