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Article

Mechanism of gemcitabine-induced suppression of human cholangiocellular carcinoma cell growth

  • Authors:
    • Yuka Toyota
    • Hisakazu Iwama
    • Kiyohito Kato
    • Joji Tani
    • Akiko Katsura
    • Miwa Miyata
    • Shintaro Fujiwara
    • Koji Fujita
    • Teppei Sakamoto
    • Takayuki Fujimori
    • Ryoichi Okura
    • Kiyoyuki Kobayashi
    • Tomoko Tadokoro
    • Shima Mimura
    • Takako Nomura
    • Hisaaki Miyoshi
    • Asahiro Morishita
    • Hideki Kamada
    • Hirohito Yoneyama
    • Keiichi Okano
    • Yasuyuki Suzuki
    • Tsutomu Masaki
  • View Affiliations / Copyright

    Affiliations: Department of Gastroenterology and Neurology, Kagawa University, Kagawa 761-0793, Japan, Life Science Research Center, Kagawa University, Kagawa 761-0793, Japan, Department of Gastroenterological Surgery, Kagawa University, Kagawa 761-0793, Japan
  • Pages: 1293-1302
    |
    Published online on: August 7, 2015
       https://doi.org/10.3892/ijo.2015.3118
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Abstract

Although gemcitabine (2',2'-difluorocytidine monohydrochloride) is a common anticancer agent of cholangiocellular carcinoma (CCC), its growth inhibitory effects and gemcitabine resistance in CCC cells are poorly understood. Our aims were to uncover the mechanism underlying the antitumor effect of gemcitabine and to analyze the mechanism regulating in vitro CCC cell gemcitabine resistance. In addition, we sought to identify miRNAs associated with the antitumor effects of gemcitabine in CCCs. Using a cell proliferation assay and flow cytometry, we examined the ability of gemcitabine to inhibit cell proliferation in three types of human CCC cell lines (HuCCT-1, Huh28, TKKK). We also employed western blotting to investigate the effects of gemcitabine on cell cycle-related molecules in CCC cells. In addition, we used array chips to assess gemcitabine-mediated changes in angiogenic molecules and activated tyrosine kinase receptors in CCC cells. We used miRNA array chips to comprehensively analyze gemcitabine-induced miRNAs and examined clusters of differentially expressed miRNAs in cells with and without gemcitabine treatment. Gemcitabine inhibited cell proliferation in a dose- and time-dependent manner in HuCCT-1 cells, whereas cell proliferation was unchanged in Huh28 and TKKK cells. Gemcitabine inhibited cell cycle progression in HuCCT-1 cells from G0/G1 to S phase, resulting in G1 cell cycle arrest due to the reduction of cyclin D1 expression. In addition, gemcitabine upregulated the angiogenic molecules IL-6, IL-8, ENA-78 and MCP-1. In TKKK cells, by contrast, gemcitabine did not arrest the cell cycle or modify angiogenic molecules. Furthermore, in gemcitabine-sensitive HuCCT-1 cells, gemcitabine markedly altered miRNA expression. The miRNAs and angiogenic molecules altered by gemcitabine contribute to the inhibition of tumor growth in vitro.
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Copy and paste a formatted citation
Spandidos Publications style
Toyota Y, Iwama H, Kato K, Tani J, Katsura A, Miyata M, Fujiwara S, Fujita K, Sakamoto T, Fujimori T, Fujimori T, et al: Mechanism of gemcitabine-induced suppression of human cholangiocellular carcinoma cell growth. Int J Oncol 47: 1293-1302, 2015.
APA
Toyota, Y., Iwama, H., Kato, K., Tani, J., Katsura, A., Miyata, M. ... Masaki, T. (2015). Mechanism of gemcitabine-induced suppression of human cholangiocellular carcinoma cell growth. International Journal of Oncology, 47, 1293-1302. https://doi.org/10.3892/ijo.2015.3118
MLA
Toyota, Y., Iwama, H., Kato, K., Tani, J., Katsura, A., Miyata, M., Fujiwara, S., Fujita, K., Sakamoto, T., Fujimori, T., Okura, R., Kobayashi, K., Tadokoro, T., Mimura, S., Nomura, T., Miyoshi, H., Morishita, A., Kamada, H., Yoneyama, H., Okano, K., Suzuki, Y., Masaki, T."Mechanism of gemcitabine-induced suppression of human cholangiocellular carcinoma cell growth". International Journal of Oncology 47.4 (2015): 1293-1302.
Chicago
Toyota, Y., Iwama, H., Kato, K., Tani, J., Katsura, A., Miyata, M., Fujiwara, S., Fujita, K., Sakamoto, T., Fujimori, T., Okura, R., Kobayashi, K., Tadokoro, T., Mimura, S., Nomura, T., Miyoshi, H., Morishita, A., Kamada, H., Yoneyama, H., Okano, K., Suzuki, Y., Masaki, T."Mechanism of gemcitabine-induced suppression of human cholangiocellular carcinoma cell growth". International Journal of Oncology 47, no. 4 (2015): 1293-1302. https://doi.org/10.3892/ijo.2015.3118
Copy and paste a formatted citation
x
Spandidos Publications style
Toyota Y, Iwama H, Kato K, Tani J, Katsura A, Miyata M, Fujiwara S, Fujita K, Sakamoto T, Fujimori T, Fujimori T, et al: Mechanism of gemcitabine-induced suppression of human cholangiocellular carcinoma cell growth. Int J Oncol 47: 1293-1302, 2015.
APA
Toyota, Y., Iwama, H., Kato, K., Tani, J., Katsura, A., Miyata, M. ... Masaki, T. (2015). Mechanism of gemcitabine-induced suppression of human cholangiocellular carcinoma cell growth. International Journal of Oncology, 47, 1293-1302. https://doi.org/10.3892/ijo.2015.3118
MLA
Toyota, Y., Iwama, H., Kato, K., Tani, J., Katsura, A., Miyata, M., Fujiwara, S., Fujita, K., Sakamoto, T., Fujimori, T., Okura, R., Kobayashi, K., Tadokoro, T., Mimura, S., Nomura, T., Miyoshi, H., Morishita, A., Kamada, H., Yoneyama, H., Okano, K., Suzuki, Y., Masaki, T."Mechanism of gemcitabine-induced suppression of human cholangiocellular carcinoma cell growth". International Journal of Oncology 47.4 (2015): 1293-1302.
Chicago
Toyota, Y., Iwama, H., Kato, K., Tani, J., Katsura, A., Miyata, M., Fujiwara, S., Fujita, K., Sakamoto, T., Fujimori, T., Okura, R., Kobayashi, K., Tadokoro, T., Mimura, S., Nomura, T., Miyoshi, H., Morishita, A., Kamada, H., Yoneyama, H., Okano, K., Suzuki, Y., Masaki, T."Mechanism of gemcitabine-induced suppression of human cholangiocellular carcinoma cell growth". International Journal of Oncology 47, no. 4 (2015): 1293-1302. https://doi.org/10.3892/ijo.2015.3118
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