Proteomic analysis of 1α,25-Dihydroxyvitamin D3 action on human colon cancer cells reveals a link to splicing regulation

14 páginas, 6 figuras, 2 tablas.El pdf del artículo es la versión pre-print.1α,25-Dihydroxyvitamin D3 (1,25(OH)2D3) and other vitamin D compounds are promising molecules in the prevention and therapy of colon cancer and other neoplasias. To study the mechanism of action of 1,25(OH)2D3 in colon cancer cells, we carried out a comparative proteomic analysis of the nuclear fractions of SW480-ADH cells treated with 1,25(OH)2D3 or vehicle during 8 or 48 h. 2D-DIGE analysis combined with MALDI-TOF-TOF mass spectrometry interrogation led to the identification of 59 differentially expressed unique proteins. Most identified proteins were nuclear, but several cytoskeleton-associated proteins were also detected. A good concordance between changes in expression at protein and RNA levels was observed for the validated proteins. A large group of identified proteins, such as SFPQ, SMARCE, KHSRP, TARDBP and PARP1, were involved in RNA processing or modification and have been ascribed to the spliceosome compartment of human cells. In addition, a smaller group of proteins (ERM (Ezrin, Radixin, Moesin) family, VCL, CORO1C, ACTB) were cytoskeleton-associated and played a role in cell adhesion and morphology. These results confirm the induction of epithelial phenotype by 1,25(OH)2D3 and suggest a role for vitamin D compounds in the regulation of the spliceosome and thus, in alternative splicing and possibly microRNA synthesis in colon cancer cells.Graphical abstract Proteomic analysis of 1α,25-Dihydroxyvitamin D3 action on human colon cancer cells reveals a link to splicing regulation. View high quality image (147K)This research was supported by grants from Colomics Programme Comunidad de Madrid (S-GEN-0266/2006), Ayudas a grupos estables de investigación Fundación Científica de la Asociación Española contra el Cáncer to J.I.C., Ministerio de Ciencia e Innovación de España (BIO2009-08818 (J.I.C.), SAF07-60341(A.M.) and SAF2010-18302 (A.M.)), Proteored platform (J.I.C.), and Red Temática de Investigación Cooperativa en Cáncer (RD06/0020/0009, ISCIII) (A.M.)Peer reviewe