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A NOVEL AND RAPID PREDICTION ASSAY FOR THE EFFECTIVENESS OF IL‐6 RECEPTOR SPECIFIC ANTISENSE OLIGONUCLEOTIDES BY PROLIFERATION INHIBITION OF AN INTERLEUKIN‐6 DEPENDENT CELL LINE
Author(s) -
Varga Lia Valeria,
Falus Andras,
Pallinger Eva,
Novák István,
Fülöp Kristóf A.,
Toth Sara,
RoseJohn Stefan
Publication year - 2001
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1006/cbir.2000.0599
Subject(s) - transfection , oligonucleotide , microbiology and biotechnology , receptor , biology , cell culture , cell growth , chemistry , gene , biochemistry , genetics
Interleukin‐6 (IL‐6) belongs to a family of cytokines that use receptors consisting of a common signal‐transducing chain (gp130). Baf/3 cells transfected with the human IL‐6 receptor (IL‐6R) and gp130 (Baf/3‐gp130/IL‐6R) can only grow in medium containing IL‐6. We attempted to interrupt the signal transducing pathway of IL‐6 with the help of antisense oligonucleotides (ASOs) designed against the IL‐6R. We used 18 different kinds of antisense oligonucleotides of overlapping sequences around the translational start codon of the human IL‐6R. Sense ASOs were used as a control. The proliferation of cells was analysed by H‐thymidine incorporation. Cell surface expression of the IL‐6R was assessed by FACS analysis. We identified three ASOs which strongly inhibited the proliferation of IL‐6 dependent transfected Baf/3 cells. Flow cytometric studies on the suppression of surface expression of IL‐6R by ASOs showed a similar pattern. These results should help to clarify the structural requirements of functionally effective ASOs in the inhibition of IL‐6R.