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Article

Enhancement of cytotoxic effects of gemcitabine by Dclk1 inhibition through suppression of Chk1 phosphorylation in human pancreatic cancer cells

  • Authors:
    • Daichi Kawamura
    • Yoshihiro Takemoto
    • Arata Nishimoto
    • Koji Ueno
    • Tohru Hosoyama
    • Bungo Shirasawa
    • Toshiki Tanaka
    • Naruji Kugimiya
    • Eijiro Harada
    • Kimikazu Hamano
  • View Affiliations / Copyright

    Affiliations: Department of Surgery and Clinical Sciences, Yamaguchi University Graduate School of Medicine, Ube, Yamaguchi 755-8505, Japan, Department of Medical Education, Yamaguchi University Graduate School of Medicine, Ube, Yamaguchi 755-8505, Japan
  • Pages: 3238-3244
    |
    Published online on: September 20, 2017
       https://doi.org/10.3892/or.2017.5974
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Abstract

Although gemcitabine (GEM) is frequently used in the treatment of pancreatic cancer, the effects are limited. To increase the inhibitory effect of GEM, the identification of a molecular target is needed. Recent studies have revealed that doublecortin-like kinase 1 (Dclk1) positively regulates tumor growth, invasion, metastasis, factors related to epithelial-mesenchymal transition (EMT), pluripotency, angiogenesis, and anti-apoptosis in pancreatic cancer cells. Therefore, Dclk1 is a potential therapeutic target for pancreatic cancer. However, the Dclk1-signaling pathway including its substrate proteins remains to be elucidated. To identify the candidate substrate proteins phosphorylated by Dclk1, we performed a cancer-related phosphorylated protein microarray using Dclk1-inhibited MIA Paca2 cells. Expression levels of phosphorylated cdc25A (p-cdc25A) and phosphorylated Chk1 (p-Chk1), belonging to the ATR pathway, were decreased by treatment with Dclk1 inhibitor LRRK2-IN-1 (LRRK), indicating Dclk1 involvement in the ATR pathway. Consistent with this finding, the GEM-induced p-Chk1 expression was significantly decreased by treatment with LRRK. Notably, combined treatment with GEM and LRRK allowed cell cycle progression without arresting at S phase, while individual treatment with GEM induced cell cycle arrest at S phase. In addition, combined treatment with GEM and LRRK increased the number of γ-H2AX-positive cells compared with that upon individual treatments. Moreover, LRRK alone, and combined treatment with GEM and LRRK, induced caspase-3 activation and PARP1 cleavage, in contrast to treatment with GEM alone. Finally, combined treatment with GEM and LRRK significantly reduced cell survival compared to individual treatment with GEM. These results indicate that Dclk1 inhibition in combination with GEM treatment offers a novel approach to treat pancreatic cancer cells.
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Copy and paste a formatted citation
Spandidos Publications style
Kawamura D, Takemoto Y, Nishimoto A, Ueno K, Hosoyama T, Shirasawa B, Tanaka T, Kugimiya N, Harada E, Hamano K, Hamano K, et al: Enhancement of cytotoxic effects of gemcitabine by Dclk1 inhibition through suppression of Chk1 phosphorylation in human pancreatic cancer cells. Oncol Rep 38: 3238-3244, 2017.
APA
Kawamura, D., Takemoto, Y., Nishimoto, A., Ueno, K., Hosoyama, T., Shirasawa, B. ... Hamano, K. (2017). Enhancement of cytotoxic effects of gemcitabine by Dclk1 inhibition through suppression of Chk1 phosphorylation in human pancreatic cancer cells. Oncology Reports, 38, 3238-3244. https://doi.org/10.3892/or.2017.5974
MLA
Kawamura, D., Takemoto, Y., Nishimoto, A., Ueno, K., Hosoyama, T., Shirasawa, B., Tanaka, T., Kugimiya, N., Harada, E., Hamano, K."Enhancement of cytotoxic effects of gemcitabine by Dclk1 inhibition through suppression of Chk1 phosphorylation in human pancreatic cancer cells". Oncology Reports 38.5 (2017): 3238-3244.
Chicago
Kawamura, D., Takemoto, Y., Nishimoto, A., Ueno, K., Hosoyama, T., Shirasawa, B., Tanaka, T., Kugimiya, N., Harada, E., Hamano, K."Enhancement of cytotoxic effects of gemcitabine by Dclk1 inhibition through suppression of Chk1 phosphorylation in human pancreatic cancer cells". Oncology Reports 38, no. 5 (2017): 3238-3244. https://doi.org/10.3892/or.2017.5974
Copy and paste a formatted citation
x
Spandidos Publications style
Kawamura D, Takemoto Y, Nishimoto A, Ueno K, Hosoyama T, Shirasawa B, Tanaka T, Kugimiya N, Harada E, Hamano K, Hamano K, et al: Enhancement of cytotoxic effects of gemcitabine by Dclk1 inhibition through suppression of Chk1 phosphorylation in human pancreatic cancer cells. Oncol Rep 38: 3238-3244, 2017.
APA
Kawamura, D., Takemoto, Y., Nishimoto, A., Ueno, K., Hosoyama, T., Shirasawa, B. ... Hamano, K. (2017). Enhancement of cytotoxic effects of gemcitabine by Dclk1 inhibition through suppression of Chk1 phosphorylation in human pancreatic cancer cells. Oncology Reports, 38, 3238-3244. https://doi.org/10.3892/or.2017.5974
MLA
Kawamura, D., Takemoto, Y., Nishimoto, A., Ueno, K., Hosoyama, T., Shirasawa, B., Tanaka, T., Kugimiya, N., Harada, E., Hamano, K."Enhancement of cytotoxic effects of gemcitabine by Dclk1 inhibition through suppression of Chk1 phosphorylation in human pancreatic cancer cells". Oncology Reports 38.5 (2017): 3238-3244.
Chicago
Kawamura, D., Takemoto, Y., Nishimoto, A., Ueno, K., Hosoyama, T., Shirasawa, B., Tanaka, T., Kugimiya, N., Harada, E., Hamano, K."Enhancement of cytotoxic effects of gemcitabine by Dclk1 inhibition through suppression of Chk1 phosphorylation in human pancreatic cancer cells". Oncology Reports 38, no. 5 (2017): 3238-3244. https://doi.org/10.3892/or.2017.5974
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