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Efficient delivery of siRNA into cytokine‐stimulated insulinoma cells silences Fas expression and inhibits Fas‐mediated apoptosis
Author(s) -
Burkhardt Brant R.,
Lyle Rachel,
Qian Keping,
Arnold Anne-Sophie,
Cheng Henrique,
Atkinson Mark A.,
Zhang Y. Clare
Publication year - 2006
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2005.12.068
Subject(s) - fas ligand , small interfering rna , gene silencing , apoptosis , fas receptor , microbiology and biotechnology , insulinoma , biology , terminal deoxynucleotidyl transferase , rna interference , cytokine , programmed cell death , cell , cancer research , chemistry , cell culture , tunel assay , immunology , transfection , rna , endocrinology , biochemistry , insulin , gene , genetics
Fas/FasL interactions have been proposed as a potentially important mechanism mediating β‐cell death in type 1 diabetes. Recent investigations suggest RNA interference, afforded by small interfering RNAs (siRNA), can provide specific and robust gene silencing in mammalian cells. The current study attempted to investigate the effects of silencing Fas expression with siRNA on Fas‐mediated apoptosis in mouse insulinoma cells following cytokine incubation. Our results indicate that siRNA is capable of rapid inhibition of cytokine‐induced Fas mRNA production and cell surface Fas protein. A complete suppression of the total Fas protein was only observed after prolonged incubation with siRNA, suggesting a slow turn‐over of Fas protein. Moreover, siRNA significantly inhibited Fas‐mediated β‐cell apoptosis assessed by Caspase‐3 and terminal deoxynucleotidyl transferase biotin‐dUTP nick end labeling assays, the extent of which positively correlated with the level of cell surface Fas. These observations provide additional evidence supporting a role for the Fas‐mediated pathway in β‐cell destruction, and suggest that siRNA targeting Fas may be of therapeutic value in preventing type 1 diabetes and improving islet cell viability in transplantation.