Investigation of the role of β-TrCP in growth hormone transduction defect (GHTD)

Eirini Kostopoulou; Andrea Paola Rojas Gil; Bessie E. Spiliotis

doi:10.1515/hmbci-2019-0029

Article

Investigation of the role of β-TrCP in growth hormone transduction defect (GHTD)

Eirini Kostopoulou , Andrea Paola Rojas Gil and Bessie E. Spiliotis

Published/Copyright: March 2, 2020

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From the journal Hormone Molecular Biology and Clinical Investigation Volume 41 Issue 2

Abstract

Background

Growth hormone(GH) and epidermal growth factor (EGF) stimulate cell growth and differentiation, and crosstalking between their signaling pathways is important for normal cellular development. Growth hormone transduction defect (GHTD) is characterized by excessive GH receptor (GHR) degradation, due to over-expression of the E3 ubiquitin ligase, cytokine inducible SH2-containing protein (CIS). GH induction of GHTD fibroblasts after silencing of messenger RNA (mRNA) CIS (siCIS) or with higher doses of GH restores normal GH signaling. β-Transducing-repeat-containing protein (β-TrCP), another E3 ubiquitin ligase, also plays a role in GHR endocytosis. We studied the role of β-TrCP in the regulation of the GH/GHR and EGF/EGF receptor (EGFR) pathways in normal and GHTD fibroblasts.

Materials and methods

Fibroblast cultures were developed from gingival biopsies of a GHTD (P) and a control child (C). Protein expression and cellular localization of β-TrCP were studied by Western immunoblotting and immunofluorescence, respectively, after: (1) GH 200 μg/L human GH (hGH) induction, either with or without silence CIS (siCIS), and (2) inductions with 200 μg/L GH or 1000 μg/L GH or 50 ng/mL EGF.

Results

After induction with: (1) GH200/siCIS, the protein expression and cytoplasmic-membrane localization of β-TrCP were increased in the patient, (2) GH200 in the control and GH1000 in the patient, the protein and cytoplasmic-membrane localization of β-TrCP were increased and (3) EGF, the protein expression and cytoplasmic-membrane localization of β-TrCP were increased in both the control and the patient.

Conclusions

(1) β-TrCP appears to be part of the negative regulatory mechanism of the GH/GHR and EGF/EGFR pathways. (2) There appears to be a negative correlation between β-TrCP and CIS. (3) In the control and GHTD patient, β-TrCP increases when CIS is suppressed, possibly as a compensatory inhibitor of the GH/GHR pathway.

Keywords: cytokine inducible SH2-containing protein/CIS; growth hormone receptor; growth impairment; ubiquitin ligase

Author statement

Research funding: Authors state no funding involved.
Conflict of interest: Authors state no conflict of interest.
Materials and Methods: Informed consent: Informed consent was provided.
Ethical approval: It has been received from the Ethics Committee of the University Hospital of Patras, Greece.

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Received: 2019-06-07

Accepted: 2019-12-16

Published Online: 2020-03-02

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Articles in the same Issue

https://doi.org/10.1515/hmbci-2019-0029

Keywords for this article

cytokine inducible SH2-containing protein/CIS; growth hormone receptor; growth impairment; ubiquitin ligase