Oncotarget

Research Papers:

FIR haplodeficiency promotes splicing to pyruvate kinase M2 in mice thymic lymphoma tissues revealed by six-plex tandem mass tag quantitative proteomic analysis

Asako Kimura, Kouichi Kitamura, Guzhanuer Ailiken, Mamoru Satoh, Toshinari Minamoto, Nobuko Tanaka, Fumio Nomura and Kazuyuki Matsushita _

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Oncotarget. 2017; 8:67955-67965. https://doi.org/10.18632/oncotarget.19061

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Abstract

Asako Kimura1, Kouichi Kitamura2,3, Guzhanuer Ailiken2, Mamoru Satoh4, Toshinari Minamoto5, Nobuko Tanaka3, Fumio Nomura4 and Kazuyuki Matsushita2,3

1Department of Medical Technology and Sciences, Narita School of Health Sciences, International University of Health and Welfare, Chiba-ken, Japan

2Department of Molecular Diagnosis, Graduate School of Medicine, Chiba University, Chiba, Japan

3Division of Laboratory Medicine, Chiba University Hospital, Chiba, Japan

4Division of Clinical Mass Spectrometry and Clinical Genetics, Chiba University Hospital, Chiba, Japan

5Division of Translational and Clinical Oncology and Surgical Oncology, Cancer Research Institute, Kanazawa University and Hospital, Kanazawa, Japan

Correspondence to:

Kazuyuki Matsushita, email: kmatsu@faculty.chiba-u.jp

Keywords: FUSE-binding protein-interacting repressor, LC-MS/MS, pyruvate kinase M2, six-plex tandem mass tags labeling, thymic lymphoma

Abbreviations: PK: pyruvate kinase; FBP: FUSE-binding protein; FIR: FBP-interacting repressor; T-ALL: T-cell type acute lymphoblastic leukemia; TMT: six-plex tandem mass tag

Received: July 20, 2016    Accepted: May 15, 2017    Published: July 07, 2017

ABSTRACT

The switch of pyruvate kinase (PK) M1 to PKM2 is pivotal for glucose metabolism in cancers. The PKM1/M2 shift is controlled by the alternative splicing of two mutually exclusive exons in the PKM gene. PKM1 is expressed in differentiated tissues, whereas PKM2 is expressed in cancer tissues. This study revealed that the haplodeficiency of FUSE-binding protein (FBP)-interacting repressor (FIR), a transcriptional repressor of the c-myc gene, contributed to the splicing of PKM1 to PKM2 in mice thymic lymphoma and/or T-cell type acute lymphoblastic leukemia (T-ALL) using six-plex tandem mass tag (TMT) quantitative proteomic analysis. TMT revealed 648 proteins that were up- or downregulated in mice thymic lymphoma tissues compared with wild type mouse. These proteins included transcription factors and proteins involved in DNA damage repair, DNA replication, T-cell activation/proliferation, apoptosis, etc. Among them, PKM2 protein, but not PKM1, was upregulated in the thymic lymphoma as well as T-ALL. Using qRT-PCR, we revealed that the activation of PKM2 mRNA was higher in thymic lymphoma cells of FIR+/−TP53−/− mice than that in control lymphocytes of FIR+/+TP53−/− sorted by flow cytometry. FIR knockdown by siRNA suppressed hnRNPA1 expression in HeLa cells. These results indicated that FIR haplodeficiency contributes the alternative splicing of PKM1 to PKM2 by partly inhibiting hnRNPA1 expression in the thymic lymphoma cells prior to T-ALL. Taken together, our findings suggest that FIR and its related spliceosomes are potential therapeutic targets for cancers, including T-ALL.


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