Melanocytes In The Role Of Glutamate Signaling Mechanism

Background:Cell cesurvival and proliferation by a number of different signal transduction pathway of regulation, to adapt to the microenvironment and extracellular stimulation. Changes in these pathways will lead to imbalance in signal transduction, causing uncontrolled cell growth. Glutamate signaling pathway is a recent discovery of another may be associated with tumor signaling pathways. Glutamate is an important central nervous system excitatory neurotransmitter, with the corresponding receptor molecules, through the cell membrane permeability changes of the cation or with A protein coupling and second messenger systems, thereby causing a complex series of signal transduction reactions. Glutamate signaling system was first identified as unique to the nervous system signal system. Which has confirmed that the signal system has been found in non-nervous system such as:bone, testis, pancreas, lung, heart and skin tissues. Some studies also show that the signal system and the nervous system and non-nervous system tumors are closely related.Glutamate receptors are divided into two categories, one is ionotropic glutamate receptors, such receptors are ligand gated ion channels, channel opening and closing control by glutamate. One NANAR is a strong voltage dependence and high Ca2+ permeability of a subtype. The other is metabotropic glutamate receptors, such receptors are a class of G protein-coupled receptors, including mGluRl can activate muscle sugar phosphate metabolism and intracellular calcium transport. Malignant evil black high, early lymph node metastasis can occur and blood metastasis, patients with short survival time, the annual incidence is rising. The bad treatment of black has been the focus of academic research. Scholars have conducted a glutamate signaling pathway in the role of the evil black.2009 have found that keratinocytes can secrete L-glutamate; and KCs with the expression of NMDAR glutamate receptor, glutamate signaling pathway may influence the intracellular calcium concentration affect keratinocyte proliferation and differentiation. Now been confirmed that the epidermis of melanoma cells also NMDAR, AMPAR and other glutamate receptors; and found that AMPA ionotropic glutamate receptor inhibitors may be reduced melanoma cells by MITF gene expression. Glutamate receptor in benign and malignant morphological changes of melanocytes, which actin and tubulin filaments disorder. Evil black and pigment disorders (such as vitiligo) in the function of melanocytes is lost by the skin of some unknown mechanism, may be associated with glutamate receptors. The need of glutamate receptors in human benign and malignant melanocytes in the role, and the glutamate receptor agonists and antagonists in benign and malignant melanocytes in morphology and growth.Objective:To investigate glutamate pathway in normal and malignant mela-nocytes in the mechanism.Method:1.Cultured human embryonic kidney cell line HEK293 cells, human evil black cell WM451LU invasive cells;adenovirus was amplified in HEK293 cells-microtubule-associated protein 2a (adenovirus-mediated MAP2a,Ad-MAP2a)2.Microtubule-associated protein 2a (Microtubule-associated protein 2a,MAP2a),ionotropic glutamate receptors in the methyl-D-aspartate receptor 2A(N-methyl-D-aspartate receptors 2A,NMDAR2A),metabotropic glutamate receptor 1 (metabotropic glutamate receptor 1,mGluR1) researchobservation of transfected Ad-MAP2a,NMDAR noncompetitive antagonist MK-801 and selective non-competitive mGluR1 antagonist CPCCOEt were under the action of In vitro migration and invasion experiments on black evil in human invasive tumor cells WM451LU role.3.Observation of transfected Ad-MAP2a,NMDAR noncompetitive antagonist MK-801 and selective non-competitive mGluRl antagonist CPCCOEt were under,through in vivo experiments in nude mice of human evil black cell WM451LU invasive tumor changes.4.Cultured primary human melanocytes. DOPA staining and electron microscope identification of melanocytes,Western Blot method of determination of primary human melanoma cells NMDAR2A expression. Scanning electron microscope and non-competitive NMDAR antagonist MK-801,agonist N-Methyl-D-aspartic acid (NMDA),Quisqualate (Q-LA) under the action of melanocyte changes.Results:1.In vitro cell migration and invasion was found that MK801, CPCCOEt could inhibit the invasiveness of human evil black cells WM451LU cell migration, invasion, and MK801, CPCCOEt,respectively MAP2a a synergistic inhibition of migration and invasion.2.Western Blot experiments revealed that the transfectants. Ad-MAP2a dark evil of human invasive cell line WM451LU cells NMDAR2A reduced expression.3.In vivo animal model of nude black evil found MK801,CPCCOEt, Ad-MAP2a could inhibit the invasiveness of human evil black WM451LU cell tumor cell line proliferation,and MK801,CPCCOEt,respectively Ad-MAP2a a synergistic inhibition of value-added role.4.Projection electron microscope showed that in vivo animal models nude black evil, MK801,CPCCOEt,Ad-MAP2a could be so evil black human invasive tumor cell lines WM451LU morphological change.5.Western Blot was found that primary human melanoma cells have NMDAR2A protein.6.Scanning electron microscope found that NMDA2R agonist NMDA and Q-LA,NMDAR inhibitor MK-801 induced melanocyte morphology can change. MCs in NMDA2R agonist NMDA and the role of Q-LA after the end of the dendritic cells significantly wider,2,3 DC increase. The role of dendritic cells after the end of MK801 significantly narrower, secondary and tertiary dendritic reduced.Conclusion:1.In benign and malignant melanoma cells exist in glutamate signaling pathway.2.In vitro malignant melanoma cells by blocking the glutamate signaling pathway can inhibit the expression of malignant melanoma, migration, and invasion.3.The body of malignant melanoma cells by blocking the glutamate signaling pathway can inhibit the malignant melanoma of the value-added.4.NMDAR glutamate receptor blockade or excitement can affect dendritic morphology of melanocytes.