Thrombopoietin stimulates migration and activates multiple signaling pathways in hepatoblastoma cells.

Roberto G Romanelli; Ilaria Petrai; Gaia Robino; Eva Efsen; Erica Novo; Andrea Bonacchi; Gabriella Pagliai; Maurizio Parola; Nadia Navari; Wanda Delogu; Francesco Vizzutti; Krista Rombouts; Paolo Gentilini; Giacomo Laffi; Fabio Marra

doi:10.1152/ajpgi.00350.2004

Thrombopoietin stimulates migration and activates multiple signaling pathways in hepatoblastoma cells.

Roberto G Romanelli, Ilaria Petrai, Gaia Robino, Eva Efsen, Erica Novo, Andrea Bonacchi, Gabriella Pagliai, Maurizio Parola, Nadia Navari, Wanda Delogu, Francesco Vizzutti, Krista Rombouts, Paolo Gentilini, Giacomo Laffi, Fabio Marra

Nutritional Immunology

21 Citations (Scopus)

Abstract

Thrombopoietin (TPO), a cytokine that participates in the differentiation and maturation of megakaryocytes, is produced in the liver, but only limited information is available on the biological response of liver-derived cells to TPO. In this study, we investigated whether HepG2 cells express c-Mpl, the receptor for TPO, and whether TPO elicits biological responses and intracellular signaling in this cell type. Specific transcripts for c-Mpl were detected in HepG2 cells by RT-PCR, and expression of the protein was demonstrated by Western blot analysis and immunofluorescence. Exposure of HepG2 cells to TPO was associated with a dose-dependent increase in cell migration and chemoinvasion through Matrigel-coated filters. A checkerboard analysis showed that the effects of TPO on cell migration were dependent on both chemotaxis and chemokinesis. Exposure of HepG2 cells to TPO resulted in the activation of different members of the MAPK family, including ERK and JNK, as assessed using phosphorylation-specific antibodies and immune complex kinase assays. TPO also activated phosphatidylinositol 3-kinase (PI3K) and the downstream kinase Akt in a time-dependent manner. Finally, activation of c-Mpl was associated with increased activation of nuclear factor-kappaB. With the use of specific inhibitors, tyrosine phosphorylation and activation of PI3K were found to be required for the induction of migration in response to TPO. We conclude that TPO exerts biological actions on cultured hepatoblastoma cells via activation of c-Mpl and its downstream signaling.

Original language	English
Journal	American Journal of Physiology: Gastrointestinal and Liver Physiology
Volume	290
Issue number	1
Pages (from-to)	G120-8
ISSN	0193-1857
DOIs	https://doi.org/10.1152/ajpgi.00350.2004
Publication status	Published - 2005

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10.1152/ajpgi.00350.2004

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Romanelli, R. G., Petrai, I., Robino, G., Efsen, E., Novo, E., Bonacchi, A., Pagliai, G., Parola, M., Navari, N., Delogu, W., Vizzutti, F., Rombouts, K., Gentilini, P., Laffi, G., & Marra, F. (2005). Thrombopoietin stimulates migration and activates multiple signaling pathways in hepatoblastoma cells. American Journal of Physiology: Gastrointestinal and Liver Physiology, 290(1), G120-8. https://doi.org/10.1152/ajpgi.00350.2004

Romanelli, RG, Petrai, I, Robino, G, Efsen, E, Novo, E, Bonacchi, A, Pagliai, G, Parola, M, Navari, N, Delogu, W, Vizzutti, F, Rombouts, K, Gentilini, P, Laffi, G & Marra, F 2005, 'Thrombopoietin stimulates migration and activates multiple signaling pathways in hepatoblastoma cells.', American Journal of Physiology: Gastrointestinal and Liver Physiology, vol. 290, no. 1, pp. G120-8. https://doi.org/10.1152/ajpgi.00350.2004

@article{466516e0acae11ddb5e9000ea68e967b,

title = "Thrombopoietin stimulates migration and activates multiple signaling pathways in hepatoblastoma cells.",

abstract = "Thrombopoietin (TPO), a cytokine that participates in the differentiation and maturation of megakaryocytes, is produced in the liver, but only limited information is available on the biological response of liver-derived cells to TPO. In this study, we investigated whether HepG2 cells express c-Mpl, the receptor for TPO, and whether TPO elicits biological responses and intracellular signaling in this cell type. Specific transcripts for c-Mpl were detected in HepG2 cells by RT-PCR, and expression of the protein was demonstrated by Western blot analysis and immunofluorescence. Exposure of HepG2 cells to TPO was associated with a dose-dependent increase in cell migration and chemoinvasion through Matrigel-coated filters. A checkerboard analysis showed that the effects of TPO on cell migration were dependent on both chemotaxis and chemokinesis. Exposure of HepG2 cells to TPO resulted in the activation of different members of the MAPK family, including ERK and JNK, as assessed using phosphorylation-specific antibodies and immune complex kinase assays. TPO also activated phosphatidylinositol 3-kinase (PI3K) and the downstream kinase Akt in a time-dependent manner. Finally, activation of c-Mpl was associated with increased activation of nuclear factor-kappaB. With the use of specific inhibitors, tyrosine phosphorylation and activation of PI3K were found to be required for the induction of migration in response to TPO. We conclude that TPO exerts biological actions on cultured hepatoblastoma cells via activation of c-Mpl and its downstream signaling.",

author = "Romanelli, {Roberto G} and Ilaria Petrai and Gaia Robino and Eva Efsen and Erica Novo and Andrea Bonacchi and Gabriella Pagliai and Maurizio Parola and Nadia Navari and Wanda Delogu and Francesco Vizzutti and Krista Rombouts and Paolo Gentilini and Giacomo Laffi and Fabio Marra",

note = "Keywords: Cell Line, Tumor; Cell Movement; Dose-Response Relationship, Drug; Enzyme Activation; Gene Expression Regulation, Neoplastic; Hepatoblastoma; Humans; Mitogen-Activated Protein Kinase Kinases; NF-kappa B; Protein Binding; Proto-Oncogene Proteins; Receptors, Cytokine; Receptors, Thrombopoietin; Signal Transduction; Thrombopoietin",

year = "2005",

doi = "10.1152/ajpgi.00350.2004",

language = "English",

volume = "290",

pages = "G120--8",

journal = "American Journal of Physiology: Gastrointestinal and Liver Physiology",

issn = "0193-1857",

publisher = "American Physiological Society",

number = "1",

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TY - JOUR

T1 - Thrombopoietin stimulates migration and activates multiple signaling pathways in hepatoblastoma cells.

AU - Romanelli, Roberto G

AU - Petrai, Ilaria

AU - Robino, Gaia

AU - Efsen, Eva

AU - Novo, Erica

AU - Bonacchi, Andrea

AU - Pagliai, Gabriella

AU - Parola, Maurizio

AU - Navari, Nadia

AU - Delogu, Wanda

AU - Vizzutti, Francesco

AU - Rombouts, Krista

AU - Gentilini, Paolo

AU - Laffi, Giacomo

AU - Marra, Fabio

N1 - Keywords: Cell Line, Tumor; Cell Movement; Dose-Response Relationship, Drug; Enzyme Activation; Gene Expression Regulation, Neoplastic; Hepatoblastoma; Humans; Mitogen-Activated Protein Kinase Kinases; NF-kappa B; Protein Binding; Proto-Oncogene Proteins; Receptors, Cytokine; Receptors, Thrombopoietin; Signal Transduction; Thrombopoietin

PY - 2005

Y1 - 2005

N2 - Thrombopoietin (TPO), a cytokine that participates in the differentiation and maturation of megakaryocytes, is produced in the liver, but only limited information is available on the biological response of liver-derived cells to TPO. In this study, we investigated whether HepG2 cells express c-Mpl, the receptor for TPO, and whether TPO elicits biological responses and intracellular signaling in this cell type. Specific transcripts for c-Mpl were detected in HepG2 cells by RT-PCR, and expression of the protein was demonstrated by Western blot analysis and immunofluorescence. Exposure of HepG2 cells to TPO was associated with a dose-dependent increase in cell migration and chemoinvasion through Matrigel-coated filters. A checkerboard analysis showed that the effects of TPO on cell migration were dependent on both chemotaxis and chemokinesis. Exposure of HepG2 cells to TPO resulted in the activation of different members of the MAPK family, including ERK and JNK, as assessed using phosphorylation-specific antibodies and immune complex kinase assays. TPO also activated phosphatidylinositol 3-kinase (PI3K) and the downstream kinase Akt in a time-dependent manner. Finally, activation of c-Mpl was associated with increased activation of nuclear factor-kappaB. With the use of specific inhibitors, tyrosine phosphorylation and activation of PI3K were found to be required for the induction of migration in response to TPO. We conclude that TPO exerts biological actions on cultured hepatoblastoma cells via activation of c-Mpl and its downstream signaling.

AB - Thrombopoietin (TPO), a cytokine that participates in the differentiation and maturation of megakaryocytes, is produced in the liver, but only limited information is available on the biological response of liver-derived cells to TPO. In this study, we investigated whether HepG2 cells express c-Mpl, the receptor for TPO, and whether TPO elicits biological responses and intracellular signaling in this cell type. Specific transcripts for c-Mpl were detected in HepG2 cells by RT-PCR, and expression of the protein was demonstrated by Western blot analysis and immunofluorescence. Exposure of HepG2 cells to TPO was associated with a dose-dependent increase in cell migration and chemoinvasion through Matrigel-coated filters. A checkerboard analysis showed that the effects of TPO on cell migration were dependent on both chemotaxis and chemokinesis. Exposure of HepG2 cells to TPO resulted in the activation of different members of the MAPK family, including ERK and JNK, as assessed using phosphorylation-specific antibodies and immune complex kinase assays. TPO also activated phosphatidylinositol 3-kinase (PI3K) and the downstream kinase Akt in a time-dependent manner. Finally, activation of c-Mpl was associated with increased activation of nuclear factor-kappaB. With the use of specific inhibitors, tyrosine phosphorylation and activation of PI3K were found to be required for the induction of migration in response to TPO. We conclude that TPO exerts biological actions on cultured hepatoblastoma cells via activation of c-Mpl and its downstream signaling.

U2 - 10.1152/ajpgi.00350.2004

DO - 10.1152/ajpgi.00350.2004

M3 - Journal article

C2 - 16150872

SN - 0193-1857

VL - 290

SP - G120-8

JO - American Journal of Physiology: Gastrointestinal and Liver Physiology

JF - American Journal of Physiology: Gastrointestinal and Liver Physiology

IS - 1

ER -

Thrombopoietin stimulates migration and activates multiple signaling pathways in hepatoblastoma cells.

Abstract

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