RESUMO
Resistance to anticancer drugs and consequent failure of chemotherapy is a complex problem severely limiting therapeutic options in metastatic cancer. Many studies have shown a role for drug efflux pumps of the ATP-binding cassette transporters family in the development of drug resistance. ClC-3, a member of the CLC family of chloride channels and transporters, is expressed in intracellular compartments of neuronal cells and involved in vesicular acidification. It has previously been suggested that acidification of intracellular organelles can promote drug resistance by increasing drug sequestration. Therefore, we hypothesized a role for ClC-3 in drug resistance. Here, we show that ClC-3 is expressed in neuroendocrine tumor cell lines, such as BON, LCC-18, and QGP-1, and localized in intracellular vesicles co-labeled with the late endosomal/lysosomal marker LAMP-1. ClC-3 overexpression increased the acidity of intracellular vesicles, as assessed by acridine orange staining, and enhanced resistance to the chemotherapeutic drug etoposide by almost doubling the IC(50) in either BON or HEK293 cell lines. Prevention of organellar acidification, by inhibition of the vacuolar H(+)-ATPase, reduced etoposide resistance. No expression of common multidrug resistance transporters, such as P-glycoprotein or multidrug-related protein-1, was detected in either the BON parental cell line or the derivative clone overexpressing ClC-3. The probable mechanism of enhanced etoposide resistance can be attributed to the increase of vesicular acidification as consequence of ClC-3 overexpression. This study therefore provides first evidence for a role of intracellular CLC proteins in the modulation of cancer drug resistance.
Assuntos
Antineoplásicos/farmacologia , Canais de Cloreto/metabolismo , Resistencia a Medicamentos Antineoplásicos , Endossomos/efeitos dos fármacos , Etoposídeo/farmacologia , Tumores Neuroectodérmicos/metabolismo , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Laranja de Acridina , Proliferação de Células/efeitos dos fármacos , Células Cultivadas/efeitos dos fármacos , Resistência a Múltiplos Medicamentos , Endossomos/metabolismo , Genes MDR/fisiologia , Humanos , Técnicas Imunoenzimáticas , Rim/efeitos dos fármacos , Proteínas Musculares/farmacologia , Tumores Neuroectodérmicos/tratamento farmacológico , Tumores Neuroectodérmicos/patologiaRESUMO
Maintenance of a constant volume is essential for normal cell function. Following cell swelling, as a consequence of reduction of extracellular osmolarity or increase of intracellular content of osmolytes, animal cells are able to restore their original volume by activation of potassium and chloride conductances. The loss of these ions, followed passively by water, is responsible for the homeostatic response called regulatory volume decrease (RVD). Activation of a chloride conductance upon cell swelling is a key step in RVD. Several proteins have been proposed as candidates for this chloride conductance. The status of the field is reviewed, with particular emphasis on ClC-3, a member of the ClC family which has been recently proposed as the chloride channel involved in cell volume regulation.
Assuntos
Tamanho Celular/fisiologia , Canais de Cloreto/fisiologia , Animais , Linhagem Celular , Canais de Cloreto/genética , Canais de Cloreto/metabolismo , Cloretos/química , Cloretos/metabolismo , Condutividade Elétrica , Humanos , Potássio/química , Potássio/metabolismo , Canais de Potássio/metabolismo , Água/metabolismo , Equilíbrio HidroeletrolíticoRESUMO
We describe the identification in aphids of a unique heterodimeric voltage-gated sodium channel which has an atypical ion selectivity filter and, unusually for insect channels, is highly insensitive to tetrodotoxin. We demonstrate that this channel has most likely arisen by adaptation (gene fission or duplication) of an invertebrate ancestral mono(hetero)meric channel. This is the only identifiable voltage-gated sodium channel homologue in the aphid genome(s), and the channel's novel selectivity filter motif (DENS instead of the usual DEKA found in other eukaryotes) may result in a loss of sodium selectivity, as indicated experimentally in mutagenised Drosophila channels.