Clostridium difficile toxin A attenuates Wnt/B-Catenin signaling in intestinal epithelial cells

Clostridium difficile toxins A and B (TcdA and TcdB) are homologous glycosyltransferases that inhibit a group of small GTPases within host cells, but several mechanisms underlying their pathogenic activity remain unclear. In this study, we evaluated the effects of TcdA on the Wnt/ -catenin pathway,...

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Detalles Bibliográficos
Autores: Lima, Bruno Bezerra, Fonseca, Bárbara Faria, Amado, Nathália da Graça, Lima, Débora Moreira, Ribeiro, Ronaldo Albuquerque, Abreu, José Garcia, Brito, Gerly Anne de Castro
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2014
País:Brasil
Institución:Universidade Federal do Ceará (UFC)
Repositorio:Repositório Institucional da Universidade Federal do Ceará (UFC)
Idioma:inglés
OAI Identifier:oai:repositorio.ufc.br:riufc/12280
Acceso en línea:http://www.repositorio.ufc.br/handle/riufc/12280
Access Level:acceso abierto
Palabra clave:Clostridium difficile
Células Epiteliais
Descripción
Sumario:Clostridium difficile toxins A and B (TcdA and TcdB) are homologous glycosyltransferases that inhibit a group of small GTPases within host cells, but several mechanisms underlying their pathogenic activity remain unclear. In this study, we evaluated the effects of TcdA on the Wnt/ -catenin pathway, the major driving force behind the proliferation of epithelial cells in colonic crypts. IEC-6 and RKO cells stimulated with Wnt3a-conditioned medium were incubated with 10, 50, and 100 ng/ml of TcdA for 24 h, resulting in a dose-dependent inhibition of the Wnt signaling, as demonstrated by a T-cell factor (TCF) reporter assay. This was further confirmed by immunofluorescence staining for nuclear localization of -catenin and Western blotting for -catenin and c-Myc (encoded by a Wnt target gene). Moreover, our Western blot analysis showed a decrease in the -catenin protein levels, which was reversed by z-VAD-fmk, a pan-caspase inhibitor. Nonetheless, TcdA was still able to inhibit the Wnt/ -catenin pathway even in the presence of z-VAD-fmk, lithium chloride (a GSK3 inhibitor), or constitutively active -catenin, as determined by a TCF reporter assay. Furthermore, preincubation of RKO cells with TcdA for 12 h also attenuated Wnt3a-mediated activation of Wnt signaling, suggesting that inactivation of Rho GTPases plays a significant role in that inhibition. Taken together, these findings suggest that attenuation of the Wnt signaling by TcdA is important for TcdA antiproliferative effects.