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Thrombopoietin upregulates nucleolin mRNA and protein in thrombopoietin-dependent megakaryocytic cell line, UT-7/TPO

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Abstract

Thrombopoietin (TPO) is a hematopoietic cytokine that regulates megakaryocytosis and thrombocytosis by binding to its receptor (c-Mpl). The signaling pathways downstream of c-Mpl include the Ras/Raf/MAP kinase and JAK/STAT pathway and are transduced into the regulation of immediate early-, early- and delayed-response genes. How these genes couple c-Mpl activation to the biochemical machinery of cell growth and cell cycle progression in hematopoietic cells is still unclear. UT-7/TPO is a recently characterized TPO-dependent cell line. Using RNA fingerprinting with arbitrarily primed PCR (RAP-PCR) to identify the TPO-regulated genes in this cell line, we found that the mRNA expression of nucleolin was upregulated in the UT-7/TPO cells in response to TPO. Concomitantly, the TPO-stimulated cells expressed an increased amount of full length nucleolin as determined by immunoblot analysis. The TPO-induced upregulation of nucleolin mRNA was not inhibited by the MEK1/2 inhibitor PD98059, suggesting that ERK/MAPK activation is not necessary for elevation of nucleolin gene expression in response to TPO in UT-7/TPO. Nucleolin is a multifunctional nucleolar protein thought to be involved in many cellular processes, including ribosome biogenesis, the processing of ribosomal RNA (rRNA), mRNA stability, transcriptional regulation, and cell proliferation. Thus, these results indicate that the upregulation of nucleolin mRNA and protein may be important for the TPO-induced effects of hematopoietic cells.

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Authors and Affiliations

  1. Japanese Red Cross, Hokkaido Red Cross Blood Center, Sapporo, Japan

    Takatoshi Ito, Mitsuhiro Fujihara, Atsushi Oda, Shinobu Wakamoto, Miki Yamaguchi, Hiroshi Azuma, Hisami Ikeda & Kenji Ikebuchi

  2. Kawasumi Laboratories Inc., Oita, Japan

    Takatoshi Ito

  3. Laboratory of Environmental Biology, Department of Preventive Medicine, Hokkaido University School of Medicine, Sapporo, Japan

    Atsushi Oda

  4. Division of Hematology, Department of Medicine, Jichi Medical School, Tochigi, Japan

    Norio Komatsu

  5. Pharmaceutical Research Laboratory, Kirin Brewery Co. Ltd., Takasaki, Japan

    Hiroshi Miyazaki

  6. Department of Biochemistry, Tokyo Medical University, Shinjyuku, Japan

    Kenji Ikebuchi

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  1. Takatoshi Ito
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Ito, T., Fujihara, M., Oda, A. et al. Thrombopoietin upregulates nucleolin mRNA and protein in thrombopoietin-dependent megakaryocytic cell line, UT-7/TPO. Mol Cell Biochem 247, 75–82 (2003). https://doi.org/10.1023/A:1024184116764

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