Significance of Plk1 regulation by miR-100 in human nasopharyngeal cancer
Wei Shi
Division of Applied Molecular Oncology, Ontario Cancer Institute, Toronto, ON, Canada
Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada
Search for more papers by this authorNehad M. Alajez
Division of Applied Molecular Oncology, Ontario Cancer Institute, Toronto, ON, Canada
Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada
Search for more papers by this authorCarlo Bastianutto
Division of Applied Molecular Oncology, Ontario Cancer Institute, Toronto, ON, Canada
Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada
Search for more papers by this authorAngela B.Y. Hui
Division of Applied Molecular Oncology, Ontario Cancer Institute, Toronto, ON, Canada
Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada
Search for more papers by this authorJoseph D. Mocanu
Division of Applied Molecular Oncology, Ontario Cancer Institute, Toronto, ON, Canada
Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada
Search for more papers by this authorEmma Ito
Division of Applied Molecular Oncology, Ontario Cancer Institute, Toronto, ON, Canada
Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada
Search for more papers by this authorKwok-Wai Lo
Department of Anatomical and Cellular Pathology, Chinese University of Hong Kong, Shatin, Hong Kong, China
Search for more papers by this authorRaymond Ng
Division of Otolaryngology-Head and Neck Surgery, Rouge Valley Health System, Toronto, ON, Canada
Search for more papers by this authorJohn Waldron
Department of Radiation Oncology, University Health Network, Toronto, ON, Canada
Department of Radiation Oncology, University of Toronto, Toronto, ON, Canada
Search for more papers by this authorBrian O'Sullivan
Department of Radiation Oncology, University Health Network, Toronto, ON, Canada
Department of Radiation Oncology, University of Toronto, Toronto, ON, Canada
Search for more papers by this authorCorresponding Author
Fei-Fei Liu
Division of Applied Molecular Oncology, Ontario Cancer Institute, Toronto, ON, Canada
Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada
Department of Radiation Oncology, University Health Network, Toronto, ON, Canada
Department of Radiation Oncology, University of Toronto, Toronto, ON, Canada
Tel: (416)-946-2123, Fax: (416)-946-4586
Department of Radiation Oncology, Princess Margaret Hospital/Ontario Cancer Institute, 610 University Avenue, Toronto, Ontario, Canada M5G 2M9Search for more papers by this authorWei Shi
Division of Applied Molecular Oncology, Ontario Cancer Institute, Toronto, ON, Canada
Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada
Search for more papers by this authorNehad M. Alajez
Division of Applied Molecular Oncology, Ontario Cancer Institute, Toronto, ON, Canada
Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada
Search for more papers by this authorCarlo Bastianutto
Division of Applied Molecular Oncology, Ontario Cancer Institute, Toronto, ON, Canada
Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada
Search for more papers by this authorAngela B.Y. Hui
Division of Applied Molecular Oncology, Ontario Cancer Institute, Toronto, ON, Canada
Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada
Search for more papers by this authorJoseph D. Mocanu
Division of Applied Molecular Oncology, Ontario Cancer Institute, Toronto, ON, Canada
Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada
Search for more papers by this authorEmma Ito
Division of Applied Molecular Oncology, Ontario Cancer Institute, Toronto, ON, Canada
Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada
Search for more papers by this authorKwok-Wai Lo
Department of Anatomical and Cellular Pathology, Chinese University of Hong Kong, Shatin, Hong Kong, China
Search for more papers by this authorRaymond Ng
Division of Otolaryngology-Head and Neck Surgery, Rouge Valley Health System, Toronto, ON, Canada
Search for more papers by this authorJohn Waldron
Department of Radiation Oncology, University Health Network, Toronto, ON, Canada
Department of Radiation Oncology, University of Toronto, Toronto, ON, Canada
Search for more papers by this authorBrian O'Sullivan
Department of Radiation Oncology, University Health Network, Toronto, ON, Canada
Department of Radiation Oncology, University of Toronto, Toronto, ON, Canada
Search for more papers by this authorCorresponding Author
Fei-Fei Liu
Division of Applied Molecular Oncology, Ontario Cancer Institute, Toronto, ON, Canada
Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada
Department of Radiation Oncology, University Health Network, Toronto, ON, Canada
Department of Radiation Oncology, University of Toronto, Toronto, ON, Canada
Tel: (416)-946-2123, Fax: (416)-946-4586
Department of Radiation Oncology, Princess Margaret Hospital/Ontario Cancer Institute, 610 University Avenue, Toronto, Ontario, Canada M5G 2M9Search for more papers by this authorAbstract
Polo-like kinase 1 (Plk1) is a critical regulator of many stages of mitosis; increasing evidence indicates that Plk1 overexpression correlates with poor clinical outcome, yet its mechanism of regulation remains unknown. Hence, a detailed evaluation was undertaken of Plk1 expression in human nasopharyngeal cancer (NPC), the cellular effects of targeting Plk1 using siRNA in combination with ionizing radiation (RT) and potential upstream microRNAs (miRs) that might regulate Plk1 expression. Using immunohistochemistry, Plk1 was observed to be overexpressed in 28 of 40 (70%) primary NPC biopsies, which in turn was associated with a higher likelihood of recurrence (p = 0.018). SiPlk1 significantly inhibited Plk1 mRNA and protein expression, and decreased Cdc25c levels in NPC cell lines. This depletion resulted in cytotoxicity of C666-1 cells, enhanced by the addition of RT, mediated by G2/M arrest, increased DNA double-strand breaks, apoptosis, and caspase activation. Immunofluorescence demonstrated that the G2/M arrest was associated with aberrant spindle formation, leading to mitotic arrest. In vivo, transfection of C666-1 cells and systemic delivery of siPlk1 decreased tumour growth. MicroRNA-100 (miR-100) was predicted to target Plk1 mRNA, which was indeed underexpressed in C666-1 cells, inversely correlating with Plk1 expression. Using luciferase constructs containing the 3′-UTR of Plk1 sequence, we document that miR-100 can directly target Plk1. Hence, our data demonstrate for the first time that underexpressed miR-100 leads to Plk1 overexpression, which in turn contributes to NPC progression. Targeting Plk1 will cause mitotic catastrophe, with significant cytotoxicity both in vitro and in vivo, underscoring the important therapeutic opportunity of Plk1 in NPC.
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