RRC ID 52406
Author Kawamura D, Takemoto Y, Nishimoto A, Ueno K, Hosoyama T, Shirasawa B, Tanaka T, Kugimiya N, Harada E, Hamano K.
Title Enhancement of cytotoxic effects of gemcitabine by Dclk1 inhibition through suppression of Chk1 phosphorylation in human pancreatic cancer cells.
Journal Oncol. Rep.
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.
Volume 38(5)
Pages 3238-3244
Published 2017-11
DOI 10.3892/or.2017.5974
PMID 29048622
MeSH Apoptosis / drug effects Ataxia Telangiectasia Mutated Proteins / genetics Benzodiazepinones / administration & dosage Cell Cycle Checkpoints / drug effects Cell Line, Tumor Cell Proliferation / drug effects Checkpoint Kinase 1 / genetics* Deoxycytidine / administration & dosage Deoxycytidine / analogs & derivatives* Epithelial-Mesenchymal Transition / drug effects Gene Expression Regulation, Neoplastic / drug effects Humans Intracellular Signaling Peptides and Proteins / antagonists & inhibitors Intracellular Signaling Peptides and Proteins / genetics* Neoplasm Proteins / genetics Pancreatic Neoplasms / drug therapy* Pancreatic Neoplasms / genetics Pancreatic Neoplasms / pathology Protein-Serine-Threonine Kinases / antagonists & inhibitors Protein-Serine-Threonine Kinases / genetics* Pyrimidines / administration & dosage Signal Transduction / drug effects
IF 3.041
Human and Animal Cells MIA Paca2(RCB2094) PANC-1(RCB2095)