Effects of the single and combined treatment with dopamine agonist, somatostatin analog and mTOR inhibitors in a human lung carcinoid cell line: an in vitro study

Claudia Pivonello, Panagoula Rousaki, Mariarosaria Negri, Maddalena Sarnataro, Maria Napolitano, Federica Zito Marino, Roberta Patalano, Maria Cristina de Martino, Concetta Sciammarella, Gaetano Rocco, Renato Franco, Gregory A. Kaltsas, Annamaria Colao, Rosario Pivonello

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Somatostatin analogues and mTOR inhibitors have been used as medical therapy in lung carcinoids with variable results. No data are available on dopamine agonists as treatment for lung carcinoids. The main aim of the current study was to evaluate the effect of the combined treatment of somatostatin analogue octreotide and the dopamine agonist cabergoline with mTOR inhibitors in an in vitro model of typical lung carcinoids: the NCI-H727 cell line. In NCI-H727 cell line, reverse transcriptase-quantitative polymerase chain reaction and immunofluorescence were assessed to characterize the expression of the somatostatin receptor 2 and 5, dopamine receptor 2 and mTOR pathway components. Fifteen typical lung carcinoids tissue samples have been used for somatostatin receptor 2, dopamine receptor 2, and the main mTOR pathway component p70S6K expression and localization by immunohistochemistry. Cell viability, fluorescence-activated cell sorting analysis and western blot have been assessed to test the pharmacological effects of octreotide, cabergoline and mTOR inhibitors, and to evaluate the activation of specific cell signaling pathways in NCI-H727 cell line. NCI-H727 cell line expressed somatostatin receptor 2, somatostatin receptor 5 and dopamine receptor 2 and all mTOR pathway components at messenger and protein levels. Somatostatin receptor 2, dopamine receptor 2, and p70S6K (non phosphorylated and phosphorylated) proteins were expressed in most typical lung carcinoids tissue samples. Octreotide and cabergoline did not reduce cell viability as single agents but, when combined with mTOR inhibitors, they potentiate mTOR inhibitors effect after long-term exposure, reducing Akt and ERK phosphorylation, mTOR escape mechanisms, and increasing the expression DNA-damage-inducible transcript 4, an mTOR suppressor. In conclusion, the single use of octreotide and cabergoline is not sufficient to block cell viability but the combined approach of these agents with mTOR inhibitors might reduce the mTOR inhibitors-induced escape mechanisms and/or activate the endogenous mTOR suppressor, potentiating the effect of the mTOR inhibitors in an in vitro model of typical lung carcinoids.

Original languageEnglish
Pages (from-to)1-18
Number of pages18
JournalEndocrine
DOIs
Publication statusAccepted/In press - Sep 29 2016

Fingerprint

Dopamine Agonists
Carcinoid Tumor
Somatostatin
Octreotide
Dopamine Receptors
Cell Line
Lung
70-kDa Ribosomal Protein S6 Kinases
Cell Survival
Therapeutics
Reverse Transcriptase Polymerase Chain Reaction
DNA Damage
Fluorescent Antibody Technique
In Vitro Techniques
Flow Cytometry
Proteins
Western Blotting
Immunohistochemistry
Phosphorylation
Pharmacology

Keywords

  • Cabergoline
  • Dopamine receptors
  • Everolimus
  • Lung carcinoids
  • mTOR pathway
  • Octreotide
  • Somatostatin analogues

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Endocrinology

Cite this

Effects of the single and combined treatment with dopamine agonist, somatostatin analog and mTOR inhibitors in a human lung carcinoid cell line : an in vitro study. / Pivonello, Claudia; Rousaki, Panagoula; Negri, Mariarosaria; Sarnataro, Maddalena; Napolitano, Maria; Marino, Federica Zito; Patalano, Roberta; de Martino, Maria Cristina; Sciammarella, Concetta; Rocco, Gaetano; Franco, Renato; Kaltsas, Gregory A.; Colao, Annamaria; Pivonello, Rosario.

In: Endocrine, 29.09.2016, p. 1-18.

Research output: Contribution to journalArticle

Pivonello, C, Rousaki, P, Negri, M, Sarnataro, M, Napolitano, M, Marino, FZ, Patalano, R, de Martino, MC, Sciammarella, C, Rocco, G, Franco, R, Kaltsas, GA, Colao, A & Pivonello, R 2016, 'Effects of the single and combined treatment with dopamine agonist, somatostatin analog and mTOR inhibitors in a human lung carcinoid cell line: an in vitro study', Endocrine, pp. 1-18. https://doi.org/10.1007/s12020-016-1079-2
Pivonello, Claudia ; Rousaki, Panagoula ; Negri, Mariarosaria ; Sarnataro, Maddalena ; Napolitano, Maria ; Marino, Federica Zito ; Patalano, Roberta ; de Martino, Maria Cristina ; Sciammarella, Concetta ; Rocco, Gaetano ; Franco, Renato ; Kaltsas, Gregory A. ; Colao, Annamaria ; Pivonello, Rosario. / Effects of the single and combined treatment with dopamine agonist, somatostatin analog and mTOR inhibitors in a human lung carcinoid cell line : an in vitro study. In: Endocrine. 2016 ; pp. 1-18.
@article{ac889bd8e8b040358ca6526710d95897,
title = "Effects of the single and combined treatment with dopamine agonist, somatostatin analog and mTOR inhibitors in a human lung carcinoid cell line: an in vitro study",
abstract = "Somatostatin analogues and mTOR inhibitors have been used as medical therapy in lung carcinoids with variable results. No data are available on dopamine agonists as treatment for lung carcinoids. The main aim of the current study was to evaluate the effect of the combined treatment of somatostatin analogue octreotide and the dopamine agonist cabergoline with mTOR inhibitors in an in vitro model of typical lung carcinoids: the NCI-H727 cell line. In NCI-H727 cell line, reverse transcriptase-quantitative polymerase chain reaction and immunofluorescence were assessed to characterize the expression of the somatostatin receptor 2 and 5, dopamine receptor 2 and mTOR pathway components. Fifteen typical lung carcinoids tissue samples have been used for somatostatin receptor 2, dopamine receptor 2, and the main mTOR pathway component p70S6K expression and localization by immunohistochemistry. Cell viability, fluorescence-activated cell sorting analysis and western blot have been assessed to test the pharmacological effects of octreotide, cabergoline and mTOR inhibitors, and to evaluate the activation of specific cell signaling pathways in NCI-H727 cell line. NCI-H727 cell line expressed somatostatin receptor 2, somatostatin receptor 5 and dopamine receptor 2 and all mTOR pathway components at messenger and protein levels. Somatostatin receptor 2, dopamine receptor 2, and p70S6K (non phosphorylated and phosphorylated) proteins were expressed in most typical lung carcinoids tissue samples. Octreotide and cabergoline did not reduce cell viability as single agents but, when combined with mTOR inhibitors, they potentiate mTOR inhibitors effect after long-term exposure, reducing Akt and ERK phosphorylation, mTOR escape mechanisms, and increasing the expression DNA-damage-inducible transcript 4, an mTOR suppressor. In conclusion, the single use of octreotide and cabergoline is not sufficient to block cell viability but the combined approach of these agents with mTOR inhibitors might reduce the mTOR inhibitors-induced escape mechanisms and/or activate the endogenous mTOR suppressor, potentiating the effect of the mTOR inhibitors in an in vitro model of typical lung carcinoids.",
keywords = "Cabergoline, Dopamine receptors, Everolimus, Lung carcinoids, mTOR pathway, Octreotide, Somatostatin analogues",
author = "Claudia Pivonello and Panagoula Rousaki and Mariarosaria Negri and Maddalena Sarnataro and Maria Napolitano and Marino, {Federica Zito} and Roberta Patalano and {de Martino}, {Maria Cristina} and Concetta Sciammarella and Gaetano Rocco and Renato Franco and Kaltsas, {Gregory A.} and Annamaria Colao and Rosario Pivonello",
year = "2016",
month = "9",
day = "29",
doi = "10.1007/s12020-016-1079-2",
language = "English",
pages = "1--18",
journal = "Endocrine",
issn = "1355-008X",
publisher = "Humana Press Inc.",

}

TY - JOUR

T1 - Effects of the single and combined treatment with dopamine agonist, somatostatin analog and mTOR inhibitors in a human lung carcinoid cell line

T2 - an in vitro study

AU - Pivonello, Claudia

AU - Rousaki, Panagoula

AU - Negri, Mariarosaria

AU - Sarnataro, Maddalena

AU - Napolitano, Maria

AU - Marino, Federica Zito

AU - Patalano, Roberta

AU - de Martino, Maria Cristina

AU - Sciammarella, Concetta

AU - Rocco, Gaetano

AU - Franco, Renato

AU - Kaltsas, Gregory A.

AU - Colao, Annamaria

AU - Pivonello, Rosario

PY - 2016/9/29

Y1 - 2016/9/29

N2 - Somatostatin analogues and mTOR inhibitors have been used as medical therapy in lung carcinoids with variable results. No data are available on dopamine agonists as treatment for lung carcinoids. The main aim of the current study was to evaluate the effect of the combined treatment of somatostatin analogue octreotide and the dopamine agonist cabergoline with mTOR inhibitors in an in vitro model of typical lung carcinoids: the NCI-H727 cell line. In NCI-H727 cell line, reverse transcriptase-quantitative polymerase chain reaction and immunofluorescence were assessed to characterize the expression of the somatostatin receptor 2 and 5, dopamine receptor 2 and mTOR pathway components. Fifteen typical lung carcinoids tissue samples have been used for somatostatin receptor 2, dopamine receptor 2, and the main mTOR pathway component p70S6K expression and localization by immunohistochemistry. Cell viability, fluorescence-activated cell sorting analysis and western blot have been assessed to test the pharmacological effects of octreotide, cabergoline and mTOR inhibitors, and to evaluate the activation of specific cell signaling pathways in NCI-H727 cell line. NCI-H727 cell line expressed somatostatin receptor 2, somatostatin receptor 5 and dopamine receptor 2 and all mTOR pathway components at messenger and protein levels. Somatostatin receptor 2, dopamine receptor 2, and p70S6K (non phosphorylated and phosphorylated) proteins were expressed in most typical lung carcinoids tissue samples. Octreotide and cabergoline did not reduce cell viability as single agents but, when combined with mTOR inhibitors, they potentiate mTOR inhibitors effect after long-term exposure, reducing Akt and ERK phosphorylation, mTOR escape mechanisms, and increasing the expression DNA-damage-inducible transcript 4, an mTOR suppressor. In conclusion, the single use of octreotide and cabergoline is not sufficient to block cell viability but the combined approach of these agents with mTOR inhibitors might reduce the mTOR inhibitors-induced escape mechanisms and/or activate the endogenous mTOR suppressor, potentiating the effect of the mTOR inhibitors in an in vitro model of typical lung carcinoids.

AB - Somatostatin analogues and mTOR inhibitors have been used as medical therapy in lung carcinoids with variable results. No data are available on dopamine agonists as treatment for lung carcinoids. The main aim of the current study was to evaluate the effect of the combined treatment of somatostatin analogue octreotide and the dopamine agonist cabergoline with mTOR inhibitors in an in vitro model of typical lung carcinoids: the NCI-H727 cell line. In NCI-H727 cell line, reverse transcriptase-quantitative polymerase chain reaction and immunofluorescence were assessed to characterize the expression of the somatostatin receptor 2 and 5, dopamine receptor 2 and mTOR pathway components. Fifteen typical lung carcinoids tissue samples have been used for somatostatin receptor 2, dopamine receptor 2, and the main mTOR pathway component p70S6K expression and localization by immunohistochemistry. Cell viability, fluorescence-activated cell sorting analysis and western blot have been assessed to test the pharmacological effects of octreotide, cabergoline and mTOR inhibitors, and to evaluate the activation of specific cell signaling pathways in NCI-H727 cell line. NCI-H727 cell line expressed somatostatin receptor 2, somatostatin receptor 5 and dopamine receptor 2 and all mTOR pathway components at messenger and protein levels. Somatostatin receptor 2, dopamine receptor 2, and p70S6K (non phosphorylated and phosphorylated) proteins were expressed in most typical lung carcinoids tissue samples. Octreotide and cabergoline did not reduce cell viability as single agents but, when combined with mTOR inhibitors, they potentiate mTOR inhibitors effect after long-term exposure, reducing Akt and ERK phosphorylation, mTOR escape mechanisms, and increasing the expression DNA-damage-inducible transcript 4, an mTOR suppressor. In conclusion, the single use of octreotide and cabergoline is not sufficient to block cell viability but the combined approach of these agents with mTOR inhibitors might reduce the mTOR inhibitors-induced escape mechanisms and/or activate the endogenous mTOR suppressor, potentiating the effect of the mTOR inhibitors in an in vitro model of typical lung carcinoids.

KW - Cabergoline

KW - Dopamine receptors

KW - Everolimus

KW - Lung carcinoids

KW - mTOR pathway

KW - Octreotide

KW - Somatostatin analogues

UR - http://www.scopus.com/inward/record.url?scp=84989180854&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84989180854&partnerID=8YFLogxK

U2 - 10.1007/s12020-016-1079-2

DO - 10.1007/s12020-016-1079-2

M3 - Article

AN - SCOPUS:84989180854

SP - 1

EP - 18

JO - Endocrine

JF - Endocrine

SN - 1355-008X

ER -