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1.
Cancer Lett ; 312(2): 197-208, 2011 Dec 22.
Artigo em Inglês | MEDLINE | ID: mdl-21917373

RESUMO

GPR4 is a member of the proton-sensing G protein-coupled receptor family. Within tumor microenvironments, the interstitial acidic pH may activate GPR4 to regulate the behavior of tumor cells. Mouse B16F10 melanoma cells and TRAMP-C1 prostate cancer cells, genetically engineered to overexpress GPR4 or the control vector, were subject to a series of cell migration, invasion and metastasis assays. Upon GPR4 overexpression and activation in an acidic pH, the migration of B16F10 and TRAMP-C1 cells was substantially inhibited in comparison to the vector control. Similar results were observed in the Matrigel invasion and transendothelial invasion assays. At the molecular level, stimulation of GPR4 by acidosis induced the activation of RhoA and the formation of actin stress fibers. In addition, treating B16F10 cells with the known Rho activator CN01 (calpeptin) strongly inhibited cell migration, recapitulating the acidosis/GPR4-induced motility inhibition phenotype. To examine the biological effects in vivo, B16F10 melanoma cells were intravenously injected into syngeneic C57BL/6 mice and pulmonary metastasis was inhibited by approximately 80% in GPR4-overexpressing B16F10 cells in comparison to the vector control. Upon treatment with the Rho activator CN01, the phenotype of the B16F10 vector cells paralleled that of the GPR4-overexpressing cells in cell migration and metastasis assays. These findings suggest that GPR4 activation by an acidic pH inhibits tumor cell migration and invasion, and the Rho GTPase is at least partly responsible for this phenotype.


Assuntos
Metástase Neoplásica/fisiopatologia , Animais , Sequência de Bases , Linhagem Celular Tumoral , Primers do DNA , Camundongos , Células NIH 3T3 , Reação em Cadeia da Polimerase , Receptores Acoplados a Proteínas G
2.
J Biophotonics ; 2(8-9): 521-7, 2009 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-19593764

RESUMO

Diffraction imaging of polystyrene spheres and B16F10 mouse melanoma cells embedded in gel has been investigated with a microscope objective. The diffraction images acquired with the objective from a sphere have been shown to be comparable to the Mie theory based projection images of the scattered light if the objective is translated to defocused positions towards the sphere. Using a confocal imaging based method to reconstruct and analyze the 3D structure, we demonstrated that genetic modifications in these cells can induce morphological changes and the modified cells can be used as an experimental model for study of the correlation between 3D morphology features and diffraction image data.


Assuntos
Melanoma/patologia , Poliestirenos/química , Espalhamento de Radiação , Animais , Linhagem Celular Tumoral , Imageamento Tridimensional , Luz , Melanoma/genética , Camundongos , Microscopia
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