Morphine-Induced Epidermal Growth Factor Pathway Activation in Non―Small Cell Lung Cancer

• Published in: Anesthesia and analgesia Vol. 113; no. 6; pp. 1353 - 1364
• Main Authors: FUJIOKA, Naomi, NGUYEN, Julia, CHUNSHENG CHEN, YUNFANG LI, PASRIJA, Teena, NIEHANS, Gloria, JOHNSON, Katherine N, GUPTA, Vinita, KRATZKE, Robert A, GUPTA, Kalpna
• Format: Journal Article
• Language: English
• Published: Hagerstown, MD Lippincott Williams & Wilkins 01.12.2011
• Subjects: Anesthesia; Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; Biological and medical sciences; Carcinoma, Non-Small-Cell Lung - enzymology; Cell Line, Transformed; Cell Line, Tumor; Epidermal Growth Factor - metabolism; Epidermal Growth Factor - physiology; Erlotinib Hydrochloride; Humans; Lung Neoplasms - enzymology; Lung Neoplasms - metabolism; MAP Kinase Signaling System - drug effects; MAP Kinase Signaling System - physiology; Medical sciences; Morphine - pharmacology; Quinazolines - pharmacology; Receptor, Epidermal Growth Factor - metabolism; Receptor, Epidermal Growth Factor - physiology; Receptors, Opioid, mu - agonists; Receptors, Opioid, mu - antagonists & inhibitors; Receptors, Opioid, mu - physiology; Lung disease; Epidermal growth factor; Respiratory disease; Opiates; Bronchus disease; Anesthesia; Morphine; Malignant tumor; Narcotic analgesic; non-small cell lung carcinoma; Cancer
• ISSN: 0003-2999; 1526-7598
• DOI: 10.1213/ane.0b013e318232b35a

Epidermal growth factor receptor (EGFR) is coactivated by the μ-opioid receptor (MOR), expressed on non-small cell lung cancer (NSCLC) cells and human lung cancer. We hypothesized that clinically used opioid analgesics that are MOR agonists coactivate EGFR, resulting in growth- and survival-promoting signaling. We used H2009, a human adenocarcinoma NSCLC cell line, with constitutive EGFR phosphorylation, which showed increased expression of MOR and the δ-opioid receptor by reverse transcriptase polymerase chain reaction. We used Western immunoblotting, magnetic bead-based Bio-Plex cytokine assay, immunofluorescent staining, BrdU incorporation enzyme-linked immunosorbent assay, and BioCoat™ Matrigel™ invasion assay to examine cell signaling, cytokine expression, colocalization of MOR and EGFR in human lung cancer, and cell proliferation and invasion, respectively. Similar to epidermal growth factor (EGF), morphine stimulated phosphorylation of EGFR, Akt/protein kinase B (Akt), and mitogen-activated protein kinase/extracellular signal regulated kinase (MAPK/ERK) signaling in H2009 cells. Opioid receptor (OR) antagonist, naloxone, EGFR tyrosine kinase inhibitor, erlotinib, and silencing of MOR and δ-opioid receptor abrogated morphine- and EGF-induced phosphorylation of signaling, suggestive of OR-mediated coactivation of EGFR. H2009 cells secreted significantly higher levels of cytokines compared with control Beas2B epithelial cells. H2009-conditioned medium stimulated MOR expression in Beas2B cells, suggesting that cytokines secreted by H2009 may be associated with increased OR expression in H2009. We observed colocalization of EGFR and MOR, in human NSCLC tissue. Functionally, morphine- and EGF-induced proliferation and invasion of H2009 cells was ameliorated by naloxone as well as erlotinib. Morphine-induced phosphorylation of EGFR occurs via ORs, leading to downstream MAPK/ERK, Akt phosphorylation, cell proliferation, and increased invasion. Notably, ORs are also associated with EGF-induced phosphorylation of EGFR. Increased coexpression of MOR and EGFR in human lung cancer suggests that morphine may have a growth-promoting effect in lung cancer.