Decrease in phospho-PRAS40 plays a role in the synergy between erlotinib and crizotinib in an EGFR and cMET wild-type squamous non-small cell lung cancer cell line

Nele van der Steen, Alessandro Leonetti, Kaylee Keller, Henk Dekker, Niccola Funel, Filip Lardon, Rob Ruijtenbeek, Marcello Tiseo, Christian Rolfo, Patrick Pauwels, Godefridus J. Peters, Elisa Giovannetti

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Introduction: Lung squamous cell carcinomas (SCC)typically harbor a strong activation of epidermal growth factor receptor (EGFR)pathway. Since one of the most common resistance mechanisms against EGFR inhibition relies on the activation of cMET parallel signaling, we investigated the efficacy of a dual blockade with erlotinib and crizotinib in EGFR and cMET wild-type lung SCC cell lines. Methods: Drug sensitivity assays were performed on LUDLU, SKMES-1, H1703, Calu1 and H520 cells. Further studies included analysis of cell cycle, apoptosis, spheroids, migration and Pathscan intracellular signaling array. Expression of emerging proteins was validated by Western blot and evaluated by immunohistochemistry in tissue-microarrays from lung cancer patients. Results: Erlotinib and crizotinib showed additive interaction in Calu1, H520 and SKMES-1, and strong synergism in the LUDLU cells (Combination Index: 0.387), associated to G2/M phase arrest, increased apoptosis, spheroid size reduction and inhibition of migration. Remarkably, this combination decreased the phosphorylation of downstream targets of MAPK and PI3K/Akt/mTOR pathways, with the largest decrease observed for PRAS40 Thr246. Moreover, it reduced the expression of both p-Her3 and p-PRAS40 in the synergistic LUDLU cells. Tissue specimens showed a higher expression of both proteins in SCC compared to adenocarcinoma histology. Conclusions: Combining erlotinib and crizotinib led to an additive/synergistic interaction in 4 out of 5 SCC cells. By combining both inhibitors, MAPK and PI3K/Akt/mTOR pathways were strongly inhibited, leading to increased cell death. p-Her3 and p-PRAS40 might be used as markers for determining the synergistic effect and for selecting potential candidates for the combination treatment.
Original languageEnglish
Pages (from-to)128-138
JournalBiochemical Pharmacology
Volume166
DOIs
Publication statusPublished - 2019

Cite this

van der Steen, Nele ; Leonetti, Alessandro ; Keller, Kaylee ; Dekker, Henk ; Funel, Niccola ; Lardon, Filip ; Ruijtenbeek, Rob ; Tiseo, Marcello ; Rolfo, Christian ; Pauwels, Patrick ; Peters, Godefridus J. ; Giovannetti, Elisa. / Decrease in phospho-PRAS40 plays a role in the synergy between erlotinib and crizotinib in an EGFR and cMET wild-type squamous non-small cell lung cancer cell line. In: Biochemical Pharmacology. 2019 ; Vol. 166. pp. 128-138.
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title = "Decrease in phospho-PRAS40 plays a role in the synergy between erlotinib and crizotinib in an EGFR and cMET wild-type squamous non-small cell lung cancer cell line",
abstract = "Introduction: Lung squamous cell carcinomas (SCC)typically harbor a strong activation of epidermal growth factor receptor (EGFR)pathway. Since one of the most common resistance mechanisms against EGFR inhibition relies on the activation of cMET parallel signaling, we investigated the efficacy of a dual blockade with erlotinib and crizotinib in EGFR and cMET wild-type lung SCC cell lines. Methods: Drug sensitivity assays were performed on LUDLU, SKMES-1, H1703, Calu1 and H520 cells. Further studies included analysis of cell cycle, apoptosis, spheroids, migration and Pathscan intracellular signaling array. Expression of emerging proteins was validated by Western blot and evaluated by immunohistochemistry in tissue-microarrays from lung cancer patients. Results: Erlotinib and crizotinib showed additive interaction in Calu1, H520 and SKMES-1, and strong synergism in the LUDLU cells (Combination Index: 0.387), associated to G2/M phase arrest, increased apoptosis, spheroid size reduction and inhibition of migration. Remarkably, this combination decreased the phosphorylation of downstream targets of MAPK and PI3K/Akt/mTOR pathways, with the largest decrease observed for PRAS40 Thr246. Moreover, it reduced the expression of both p-Her3 and p-PRAS40 in the synergistic LUDLU cells. Tissue specimens showed a higher expression of both proteins in SCC compared to adenocarcinoma histology. Conclusions: Combining erlotinib and crizotinib led to an additive/synergistic interaction in 4 out of 5 SCC cells. By combining both inhibitors, MAPK and PI3K/Akt/mTOR pathways were strongly inhibited, leading to increased cell death. p-Her3 and p-PRAS40 might be used as markers for determining the synergistic effect and for selecting potential candidates for the combination treatment.",
author = "{van der Steen}, Nele and Alessandro Leonetti and Kaylee Keller and Henk Dekker and Niccola Funel and Filip Lardon and Rob Ruijtenbeek and Marcello Tiseo and Christian Rolfo and Patrick Pauwels and Peters, {Godefridus J.} and Elisa Giovannetti",
year = "2019",
doi = "10.1016/j.bcp.2019.05.014",
language = "English",
volume = "166",
pages = "128--138",
journal = "Biochemical Pharmacology",
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Decrease in phospho-PRAS40 plays a role in the synergy between erlotinib and crizotinib in an EGFR and cMET wild-type squamous non-small cell lung cancer cell line. / van der Steen, Nele; Leonetti, Alessandro; Keller, Kaylee; Dekker, Henk; Funel, Niccola; Lardon, Filip; Ruijtenbeek, Rob; Tiseo, Marcello; Rolfo, Christian; Pauwels, Patrick; Peters, Godefridus J.; Giovannetti, Elisa.

In: Biochemical Pharmacology, Vol. 166, 2019, p. 128-138.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Decrease in phospho-PRAS40 plays a role in the synergy between erlotinib and crizotinib in an EGFR and cMET wild-type squamous non-small cell lung cancer cell line

AU - van der Steen, Nele

AU - Leonetti, Alessandro

AU - Keller, Kaylee

AU - Dekker, Henk

AU - Funel, Niccola

AU - Lardon, Filip

AU - Ruijtenbeek, Rob

AU - Tiseo, Marcello

AU - Rolfo, Christian

AU - Pauwels, Patrick

AU - Peters, Godefridus J.

AU - Giovannetti, Elisa

PY - 2019

Y1 - 2019

N2 - Introduction: Lung squamous cell carcinomas (SCC)typically harbor a strong activation of epidermal growth factor receptor (EGFR)pathway. Since one of the most common resistance mechanisms against EGFR inhibition relies on the activation of cMET parallel signaling, we investigated the efficacy of a dual blockade with erlotinib and crizotinib in EGFR and cMET wild-type lung SCC cell lines. Methods: Drug sensitivity assays were performed on LUDLU, SKMES-1, H1703, Calu1 and H520 cells. Further studies included analysis of cell cycle, apoptosis, spheroids, migration and Pathscan intracellular signaling array. Expression of emerging proteins was validated by Western blot and evaluated by immunohistochemistry in tissue-microarrays from lung cancer patients. Results: Erlotinib and crizotinib showed additive interaction in Calu1, H520 and SKMES-1, and strong synergism in the LUDLU cells (Combination Index: 0.387), associated to G2/M phase arrest, increased apoptosis, spheroid size reduction and inhibition of migration. Remarkably, this combination decreased the phosphorylation of downstream targets of MAPK and PI3K/Akt/mTOR pathways, with the largest decrease observed for PRAS40 Thr246. Moreover, it reduced the expression of both p-Her3 and p-PRAS40 in the synergistic LUDLU cells. Tissue specimens showed a higher expression of both proteins in SCC compared to adenocarcinoma histology. Conclusions: Combining erlotinib and crizotinib led to an additive/synergistic interaction in 4 out of 5 SCC cells. By combining both inhibitors, MAPK and PI3K/Akt/mTOR pathways were strongly inhibited, leading to increased cell death. p-Her3 and p-PRAS40 might be used as markers for determining the synergistic effect and for selecting potential candidates for the combination treatment.

AB - Introduction: Lung squamous cell carcinomas (SCC)typically harbor a strong activation of epidermal growth factor receptor (EGFR)pathway. Since one of the most common resistance mechanisms against EGFR inhibition relies on the activation of cMET parallel signaling, we investigated the efficacy of a dual blockade with erlotinib and crizotinib in EGFR and cMET wild-type lung SCC cell lines. Methods: Drug sensitivity assays were performed on LUDLU, SKMES-1, H1703, Calu1 and H520 cells. Further studies included analysis of cell cycle, apoptosis, spheroids, migration and Pathscan intracellular signaling array. Expression of emerging proteins was validated by Western blot and evaluated by immunohistochemistry in tissue-microarrays from lung cancer patients. Results: Erlotinib and crizotinib showed additive interaction in Calu1, H520 and SKMES-1, and strong synergism in the LUDLU cells (Combination Index: 0.387), associated to G2/M phase arrest, increased apoptosis, spheroid size reduction and inhibition of migration. Remarkably, this combination decreased the phosphorylation of downstream targets of MAPK and PI3K/Akt/mTOR pathways, with the largest decrease observed for PRAS40 Thr246. Moreover, it reduced the expression of both p-Her3 and p-PRAS40 in the synergistic LUDLU cells. Tissue specimens showed a higher expression of both proteins in SCC compared to adenocarcinoma histology. Conclusions: Combining erlotinib and crizotinib led to an additive/synergistic interaction in 4 out of 5 SCC cells. By combining both inhibitors, MAPK and PI3K/Akt/mTOR pathways were strongly inhibited, leading to increased cell death. p-Her3 and p-PRAS40 might be used as markers for determining the synergistic effect and for selecting potential candidates for the combination treatment.

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UR - https://www.ncbi.nlm.nih.gov/pubmed/31078602

U2 - 10.1016/j.bcp.2019.05.014

DO - 10.1016/j.bcp.2019.05.014

M3 - Article

VL - 166

SP - 128

EP - 138

JO - Biochemical Pharmacology

JF - Biochemical Pharmacology

SN - 0006-2952

ER -