EFECTOS CITOTÓXICOS DE LAS COMBINACIONES DE LA AURANOFINA CON DOS INHIBIDORES DE LA PROTEÍNA QUINASA CK2 EN UNA LÍNEA CELULAR DE LINFOCITOS T :: CYTOTOXIC EFFECTS OF THE COMBINATIONS OF AURANOFIN WITH TWO INHIBITORS OF PROTEIN KINASE CK2 INHIBITORS IN A T-CELL LINE

Marla Paredes, Germán Fraile, Mait Velásquez, Limnar Rodríguez, José Duque-Rivera

Resumen


La auranofina (AF) es una droga antiartrítica que exhibe actividad antitumoral. Mediante la inhibición de la actividad de la tiorredoxina reductasa (TrxR) y el incremento del estrés oxidativo, la AF puede inducir la muerte de células malignas aunque también es capaz de activar el factor de transcripción de enzimas antioxidantes Nrf2, que limita su eficacia. En vista de que la óptima activación de Nrf2 requiere de fosforilación por quinasas tales como la proteína quinasa CK2, en este trabajo se evaluaron, en las células Jurkat, los efectos de las combinaciones de la AF con dos inhibidores de la CK2: emodina (EM) y 4,5,6,7-tetrabromo-1H-benzotriazol (TBB) mediante el empleo de relaciones molares fijas y posterior análisis con el método de Chou-Talalay. La actividad de la TrxR fue potentemente inhibida por la AF a concentraciones nanomolares. El pretratamiento con catalasa seguido por incubación con AF 1 µM revirtió la pérdida de viabilidad inducida por la droga sola. A la misma concentración, la AF incrementó el nivel de tioles totales intracelulares, mientras que la inhibición de la síntesis del GSH con butionina sulfoximina seguida por incubación con la AF, aumentó la muerte celular. Esos resultados sugirieron que la AF estimula una respuesta adaptativa al estrés oxidativo en las células Jurkat. Las combinaciones AF/EM (1/12) y AF/TBB (1/50) inhibieron la proliferación e indujeron muerte celular de un modo sinérgico.

Palabras clave: Células Jurkat, emodina, TBB, TrxR, sinergia.

 

ABSTRACT

Auranofin (AF) is an antiarthritic drug with potential anticancer activity. By inhibiting thioredoxin reductase (TrxR) activity and increasing intracellular reactive oxygen species levels, AF can induce a lethal oxidative stress response in malignant cells. However, AF can activate the antioxidant transcription factor Nrf2 which may limit its efficacy. Because phosphorylation by kinases such as CK2 is essential for the full activation of Nrf2, the combinations of AF with the CK2 inhibitors emodin (EM) and TBB, were evaluated in Jurkat T cells by using a fixed dose ratio according to the Chou-Talalay method. The TrxR activity was potently inhibited by low nanomolar concentrations of AF. Moreover, by treatment with AF 1 µM, the observed decrease of cell viability was partly reversed by pretreatment with catalase. At the same concentration, AF increased the total thiols whereas inhibition of GSH synthesis with buthionine sulfoximine, followed by incubation with AF, resulted in stronger inhibition of viability. These results suggested that AF stimulates an adaptive response to oxidative stress in Jurkat cells. Essentially, the combinations of AF with EM (molar ratio AF/EM = 1/12) or TBB (molar ratio AF/TBB = 1/50) synergistically inhibited cell proliferation and induced cell death.

Key words: Jurkat cells, emodin, TBB, TrxR, synergy.


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