transforming-growth-factor-alpha and Nasopharyngeal-Neoplasms

In clinical practice, the common strategy for immunotherapy of nasopharyngeal carcinoma (NPC) is to infuse cytotoxic T-lymphocyte (CTL) lines several times by intravenous injection, but it is difficult by laboratory research to investigate the relationship between treatment time-point, the amount of CTL added and the therapeutic effect. The objective of this study is to establish a mathematical model to study the therapeutic effect of different treatment time-points and amounts of CTL, and to predict the change in therapeutic effect when the percentage of EBV LMP2-specific CTL is increased from 10% to 20%.. The concentration of epidermal growth factor receptor (EGFR) in the tumor cell cytomembranes increases after CTL is added. Concurrently, there is a marked downward trend of the phosphorylated transforming growth factor-α (TGFα)-EGFR complex in the tumor cell cytomembranes, which indicates restriction of tumor growth after CTL immunotherapy. The relationships among the time of addition of CTL, the amount of CTL added, different CTL specificities for LMP2 and the increment rate k of the total number of tumor cells were evaluated.. The simulation results quantify the relationships among treatment time-points, amount of CTL added, and the corresponding therapeutic effect of immunotherapy for NPC.

Topics: Carcinoma; Computer Simulation; ErbB Receptors; Humans; Immunotherapy; Models, Immunological; Nasopharyngeal Carcinoma; Nasopharyngeal Neoplasms; Phosphorylation; Reproducibility of Results; T-Lymphocytes, Cytotoxic; Time Factors; Transforming Growth Factor alpha; Viral Matrix Proteins

Although several groups had conducted proteomics of nasopharyngeal carcinoma (NPC), little study was involved in phosphoproteomics, oncogenic signaling, and cancer research about NPC. Analysis of phosphotyrosine proteins from transforming growth factor alpha (TGF-a) triggered phosphotyrosine proteome permitted the identification of novel downstream substrates of the epidermal growth factor receptor (EGFR). Using functional proteomics technology based on 2-DE, 2-D western blotting, and mass spectrometry, we identified and quantified the tyrosine phosphorylation levels of 16 proteins between control and TGF-a-treated CNE2 human NPC cells. Among these proteins, tyrosine phosphorylated levels of ten proteins were increased, and those of six proteins were decreased in TGF-a-treated CNE2 cells compared with control. In addition, among these identified proteins, ANXA3, KRT8, and KRT18 were validated to be novel tyrosine-phosphorylation targets of EGFR signaling by IP-western blotting and part of a complex EGFR phosphotyrosine signaling network. These novel findings will provide new insights into the complex EGFR phosphorylation signaling and may have implications in molecular cancer therapy of NPC.

Topics: Aged; Amino Acid Sequence; Annexin A3; Biomarkers, Tumor; Blotting, Western; Cell Differentiation; Electrophoresis, Gel, Two-Dimensional; ErbB Receptors; Humans; Keratin-18; Keratin-8; Male; Molecular Sequence Data; Nasopharyngeal Neoplasms; Phosphoproteins; Phosphorylation; Phosphotyrosine; Prognosis; Proteomics; Sequence Homology, Amino Acid; Signal Transduction; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Transforming Growth Factor alpha; Tumor Cells, Cultured

The epidermal growth factor receptor (EGFR) is usually overexpressed in nasopharyngeal carcinoma (NPC) and is associated with pathogenesis of NPC. However, while EGFR-modulated intracellular proteins have been extensively studied, little is known concerning their extracellular counterparts. To identify EGFR-regulated secreted proteins in NPC, we compared the secretome profiles of TGF-α-stimulated and unstimulated NPC cell line CNE-2. CNE-2 cells were cultured in the absence or presence of TGF-α for 24 h, and secreted proteins were obtained from conditioned serum-free media and enriched by ultrafiltration centrifugation. Using 2-DE and subsequent mass spectrometry, we identified 16 differential secreted proteins, among which the amyloid β-protein precursor (APP) was up-regulated and cystatin C was down-regulated after TGF-α stimulation. We further showed that the secretory changes of APP and cystatin C in CNE-2 after TGF-α stimulation could be abrogated by pretreatment of EGFR tyrosine kinase inhibitor PD153035 and PI3 kinase inhibitor Wortmannin, validating that APP and cystatin C are EGFR-regulated secreted proteins in NPC cells. Immunohistochemistry showed that the expression level of EGFR was positively correlated with the expression level of APP and negatively correlated with the expression level of cystatin C in NPC tissues, indicating that EGFR also regulates expression of APP and cystatin C in clinical NPC tissues. Furthermore, functional analysis showed that the growth and migration of CNE-2 cells decreased after neutralization of secretory APP in the medium using the anti-APP antibody. Our data provide substantial evidence that APP and cystatin C are target secreted proteins of EGFR in NPC, and upregulation of secretory APP by EGFR may be involved in the pathogenesis of NPC.

Topics: Amyloid beta-Protein Precursor; Androstadienes; Antibodies, Monoclonal; Blotting, Western; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cystatin C; Electrophoresis, Gel, Two-Dimensional; Enzyme Inhibitors; ErbB Receptors; Humans; Immunohistochemistry; Nasopharyngeal Neoplasms; Proteomics; Quinazolines; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Transforming Growth Factor alpha; Wortmannin

To determine serum levels of soluble intercellular adhesion molecule 1 (sICAM-1) and transforming growth factor alpha (TGF-alpha) in 51 patients with nasopharyngeal carcinoma before, during, and after radiation therapy (5-year follow-up period).. The mean +/- SD serum levels of the 2 cytokines were found to be higher in patients before radiotherapy (sICAM-1, 369.6 +/- 123.7 ng/mL; TGF-alpha, 36.6 +/- 24.6 ng/mL) than after radiotherapy (sICAM-1, 225.9 +/- 124.3 ng/mL; TGF-alpha, 20.2 +/- 22.3 ng/mL) (P<.05), and they were significantly higher in patients with recurrence (sICAM-1, 512.5 +/- 271.2 ng/mL; TGF-alpha, 48.2 +/- 23.4 ng/mL) and in those who died (sICAM-1, 542.6 +/- 245.4 ng/mL; TGF-alpha, 50.2 +/- 28.8 ng/mL) than in patients with no recurrence (sICAM-1, 217.9 +/- 116.4 ng/mL; TGF-d, 21.5 +/- 26.8 ng/mL) and in those who survived (sICAM-1, 209.4 +/- 167.2 ng/mL; TGF-alpha, 20.4 +/- 27.3 ng/mL) (P<.05). The increases in serum levels occurred approximately 3 months before relapse.. We found that sICAM-1 and TGF-alpha levels are extremely useful markers for predicting illness, recurrence, and survival.

Topics: Carcinoma; Case-Control Studies; Female; Follow-Up Studies; Humans; Intercellular Adhesion Molecule-1; Male; Middle Aged; Nasopharyngeal Neoplasms; Neoplasm Recurrence, Local; Predictive Value of Tests; Prognosis; Transforming Growth Factor alpha