ABSTRACT
Title
Cortagine, a selective CRH-R1 agonist, inhibits cell proliferation of human endometrial adenocarcinoma cells
Authors
F. Miceli1, A. Muzi2, E. Cantalupo1.
1Institute of Pharmacology, CatholicUniversityMedicalSchool, 00168 Rome, Italy
2Department of Neuroscience, University of Rome“Tor Vergata”, 00133 Rome, Italy
1Institute of Pharmacology, CatholicUniversityMedicalSchool, 00168 Rome, Italy
2Department of Neuroscience, University of Rome“Tor Vergata”, 00133 Rome, Italy
Abstract
BACKGROUND:
Corticotrophin-releasing hormone (CRH) is a 41-amino acid peptide originally characterized as a hypothalamic factor mediating neuroendocrine responses to stress.During the last years, several studies have shown that CRF is also produced by various peripheral tissues. CRH and its receptors are expressed in human normal endometrial cells, where they are associated to anti-proliferative progesterone-like activity. The gene expression and biosynthesis of CRH have also been demonstrated in Ishikawa (IK) cells, a tumor cell line derived from human endometrium.
CRH also exhibits an anti-proliferative effect on IK cells expressing CRH-R1. Since endogenous selective ligands for CRH-R1 are not yet known we used Cortagine, a newly developed selective CRH-R1 peptide agonist, to characterize specific CRH1-mediated functions in IK cells focusing on cell proliferation.
MATERIALS AND METHODS:
The cells were treated with graded concentration of Cortagine (10-7-10-10M) and with medium alone or containing drug solvent, were than incubated at 37 Cfor 5 days, adding the peptide each day. Cell growth was evaluated every 24h by counting cells in quadruplicate with the NucleoCounter (Chemometec), an automatic cell counting device. Cell proliferation was also assessed with the MTS Cell Proliferation Assay kit according to the manufacturer's instructions. This assay is a commercial cell viability assays based on the measurement of cell metabolism (CellTiter 96® AQueous One Solution Cell Proliferation Assay - MTS)
RESULTS:
Cortagine was able to inhibit cell proliferation, with respect to untreated or vehicle-treated controls, in a time- and dose-dependent manner. CRH inhibitory activity was confirmed using MTS assay. The maximum inhibition was observed with Cortagine 100 nM at 24h, the same concentration significantly inhibited cell growth after 48 and 72 h of incubation.
CONCLUSION:
In conclusion Cortagine has been shown to possess growth inhibitory effetcs on Ishikawa tumor cell line, accordingly with our previous results obtained with CRH and we have confirmed that the activation of CRH-R1 receptor by the ligand contributes to reducing cell proliferation.
The relevance of the present findings in human pathology needs to be further verified since a CRH-R1 agonist could be evaluated as targeted therapy for inhibition of tumor cell proliferation in human endometrial cancer.
Corticotrophin-releasing hormone (CRH) is a 41-amino acid peptide originally characterized as a hypothalamic factor mediating neuroendocrine responses to stress.During the last years, several studies have shown that CRF is also produced by various peripheral tissues. CRH and its receptors are expressed in human normal endometrial cells, where they are associated to anti-proliferative progesterone-like activity. The gene expression and biosynthesis of CRH have also been demonstrated in Ishikawa (IK) cells, a tumor cell line derived from human endometrium.
CRH also exhibits an anti-proliferative effect on IK cells expressing CRH-R1. Since endogenous selective ligands for CRH-R1 are not yet known we used Cortagine, a newly developed selective CRH-R1 peptide agonist, to characterize specific CRH1-mediated functions in IK cells focusing on cell proliferation.
MATERIALS AND METHODS:
The cells were treated with graded concentration of Cortagine (10-7-10-10M) and with medium alone or containing drug solvent, were than incubated at 37 Cfor 5 days, adding the peptide each day. Cell growth was evaluated every 24h by counting cells in quadruplicate with the NucleoCounter (Chemometec), an automatic cell counting device. Cell proliferation was also assessed with the MTS Cell Proliferation Assay kit according to the manufacturer's instructions. This assay is a commercial cell viability assays based on the measurement of cell metabolism (CellTiter 96® AQueous One Solution Cell Proliferation Assay - MTS)
RESULTS:
Cortagine was able to inhibit cell proliferation, with respect to untreated or vehicle-treated controls, in a time- and dose-dependent manner. CRH inhibitory activity was confirmed using MTS assay. The maximum inhibition was observed with Cortagine 100 nM at 24h, the same concentration significantly inhibited cell growth after 48 and 72 h of incubation.
CONCLUSION:
In conclusion Cortagine has been shown to possess growth inhibitory effetcs on Ishikawa tumor cell line, accordingly with our previous results obtained with CRH and we have confirmed that the activation of CRH-R1 receptor by the ligand contributes to reducing cell proliferation.
The relevance of the present findings in human pathology needs to be further verified since a CRH-R1 agonist could be evaluated as targeted therapy for inhibition of tumor cell proliferation in human endometrial cancer.