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Interferon-alpha resistance in renal carcinoma cells is associated with defective induction of signal transducer and activator of transcription 1 which can be restored by a supernatant of phorbol 12-myristate 13-acetate stimulated peripheral blood mononuclear cells.

Brinckmann A, Axer S, Jakschies D, Dallmann I, Grosse J, Patzelt T, Bernier T, Emmendoerffer A, Atzpodien J - Br. J. Cancer (2002)

Bottom Line: Therapy of selected human malignancies with interferon-alpha is widely accepted but often complicated by the emergence of interferon-alpha resistance.Interferon is a pleiotropic cytokine with antiproliferative, antitumour, antiviral and immunmodulatory effect; it signals through the Jak-STAT signal transduction pathway where signal transducer and activator of transcription 1 plays an important role.Preliminary experiments on the identification of the molecules that reinducing signal transducers and activators of transcription 1 indicate that interferon-gamma may be the responsible candidate cytokine, but several others may be involved as well.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology and Oncology, Medizinische Hochschule, Hannover, Germany.

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Electrophoretic mobility shift assays for STAT1 induction in A-498 cells by PMA-stimulated PBMC supernatant. (A) Electrophoretic mobility shift assay for STAT1 induction in IFN-α sensitive (S) A-498 cells by PMA-stimulated PBMC supernatant. Cells were either left untreated (lane 1) or treated with IFN-α alone (lane 2) or supernatant alone (lanes 3, 5, 7, 9) or with their combination (lanes 4, 6, 8 and 10). STAT1 is induced in the presence of IFN-α (lanes 2, 4, 6, 8 and 10) and after incubation for 1 h with the supernatant (lane 3) (see arrow for STAT1 band). (B) STAT1 induction in IFN-α resistant (R) A-498 cells by PMA-stimulated PBMC supernatant. Cells were treated with IFN-α alone (lane 3) or supernatant alone (lanes 4, 6, 8 and 10) or with their combination (lanes 5, 7, 9 and 11). IFN-α does not induce STAT1 (lane 3) but treatment with supernatant alone and in combination with IFN-α for 1, 4 and 6 h can induce STAT1 induction (lanes 4–9). IFN-α treated A-498 S cells served as positive control (lane 1) (see arrow for STAT1 band).
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fig2: Electrophoretic mobility shift assays for STAT1 induction in A-498 cells by PMA-stimulated PBMC supernatant. (A) Electrophoretic mobility shift assay for STAT1 induction in IFN-α sensitive (S) A-498 cells by PMA-stimulated PBMC supernatant. Cells were either left untreated (lane 1) or treated with IFN-α alone (lane 2) or supernatant alone (lanes 3, 5, 7, 9) or with their combination (lanes 4, 6, 8 and 10). STAT1 is induced in the presence of IFN-α (lanes 2, 4, 6, 8 and 10) and after incubation for 1 h with the supernatant (lane 3) (see arrow for STAT1 band). (B) STAT1 induction in IFN-α resistant (R) A-498 cells by PMA-stimulated PBMC supernatant. Cells were treated with IFN-α alone (lane 3) or supernatant alone (lanes 4, 6, 8 and 10) or with their combination (lanes 5, 7, 9 and 11). IFN-α does not induce STAT1 (lane 3) but treatment with supernatant alone and in combination with IFN-α for 1, 4 and 6 h can induce STAT1 induction (lanes 4–9). IFN-α treated A-498 S cells served as positive control (lane 1) (see arrow for STAT1 band).

Mentions: Electrophoretic mobility shift assays for IFN-α induced STAT1 induction. IFN-α sensitive (S) and IFN-α resistant (R) A-498 and fresh RCC cells and Caki-2 cells were either left untreated or incubated with 1000 IU ml−1 IFN-α for 30 min. Lanes 1–5 are A-498 cells: lanes 1 and 4, untreated IFN-α sensitive and resistant cells, respectively; lanes 2, 3 and 5, IFN-α stimulated IFN-α sensitive and resistant cells, respectively. Lanes 6 and 7 are Caki-2 cells: lane 6, untreated Caki-2 cells; lane 7, IFN-α stimulated cells. Lanes 8–11 are fresh RCC cells: lanes 8 and 10, untreated IFN-α sensitive and resistant cells, respectively; lanes 9 and 11, IFN-α stimulated IFN-α sensitive and resistant cells, respectively. STAT1 induction can be detected in sensitive A-498 and fresh RCC cells after stimulation with IFN-α but not in resistant cells (lanes 2 and 9). STAT1 can be competed with monoclonal anti-STAT1 antibody (lane 3) (see arrow for STAT1 band).


Interferon-alpha resistance in renal carcinoma cells is associated with defective induction of signal transducer and activator of transcription 1 which can be restored by a supernatant of phorbol 12-myristate 13-acetate stimulated peripheral blood mononuclear cells.

Brinckmann A, Axer S, Jakschies D, Dallmann I, Grosse J, Patzelt T, Bernier T, Emmendoerffer A, Atzpodien J - Br. J. Cancer (2002)

Electrophoretic mobility shift assays for STAT1 induction in A-498 cells by PMA-stimulated PBMC supernatant. (A) Electrophoretic mobility shift assay for STAT1 induction in IFN-α sensitive (S) A-498 cells by PMA-stimulated PBMC supernatant. Cells were either left untreated (lane 1) or treated with IFN-α alone (lane 2) or supernatant alone (lanes 3, 5, 7, 9) or with their combination (lanes 4, 6, 8 and 10). STAT1 is induced in the presence of IFN-α (lanes 2, 4, 6, 8 and 10) and after incubation for 1 h with the supernatant (lane 3) (see arrow for STAT1 band). (B) STAT1 induction in IFN-α resistant (R) A-498 cells by PMA-stimulated PBMC supernatant. Cells were treated with IFN-α alone (lane 3) or supernatant alone (lanes 4, 6, 8 and 10) or with their combination (lanes 5, 7, 9 and 11). IFN-α does not induce STAT1 (lane 3) but treatment with supernatant alone and in combination with IFN-α for 1, 4 and 6 h can induce STAT1 induction (lanes 4–9). IFN-α treated A-498 S cells served as positive control (lane 1) (see arrow for STAT1 band).
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Related In: Results  -  Collection

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fig2: Electrophoretic mobility shift assays for STAT1 induction in A-498 cells by PMA-stimulated PBMC supernatant. (A) Electrophoretic mobility shift assay for STAT1 induction in IFN-α sensitive (S) A-498 cells by PMA-stimulated PBMC supernatant. Cells were either left untreated (lane 1) or treated with IFN-α alone (lane 2) or supernatant alone (lanes 3, 5, 7, 9) or with their combination (lanes 4, 6, 8 and 10). STAT1 is induced in the presence of IFN-α (lanes 2, 4, 6, 8 and 10) and after incubation for 1 h with the supernatant (lane 3) (see arrow for STAT1 band). (B) STAT1 induction in IFN-α resistant (R) A-498 cells by PMA-stimulated PBMC supernatant. Cells were treated with IFN-α alone (lane 3) or supernatant alone (lanes 4, 6, 8 and 10) or with their combination (lanes 5, 7, 9 and 11). IFN-α does not induce STAT1 (lane 3) but treatment with supernatant alone and in combination with IFN-α for 1, 4 and 6 h can induce STAT1 induction (lanes 4–9). IFN-α treated A-498 S cells served as positive control (lane 1) (see arrow for STAT1 band).
Mentions: Electrophoretic mobility shift assays for IFN-α induced STAT1 induction. IFN-α sensitive (S) and IFN-α resistant (R) A-498 and fresh RCC cells and Caki-2 cells were either left untreated or incubated with 1000 IU ml−1 IFN-α for 30 min. Lanes 1–5 are A-498 cells: lanes 1 and 4, untreated IFN-α sensitive and resistant cells, respectively; lanes 2, 3 and 5, IFN-α stimulated IFN-α sensitive and resistant cells, respectively. Lanes 6 and 7 are Caki-2 cells: lane 6, untreated Caki-2 cells; lane 7, IFN-α stimulated cells. Lanes 8–11 are fresh RCC cells: lanes 8 and 10, untreated IFN-α sensitive and resistant cells, respectively; lanes 9 and 11, IFN-α stimulated IFN-α sensitive and resistant cells, respectively. STAT1 induction can be detected in sensitive A-498 and fresh RCC cells after stimulation with IFN-α but not in resistant cells (lanes 2 and 9). STAT1 can be competed with monoclonal anti-STAT1 antibody (lane 3) (see arrow for STAT1 band).

Bottom Line: Therapy of selected human malignancies with interferon-alpha is widely accepted but often complicated by the emergence of interferon-alpha resistance.Interferon is a pleiotropic cytokine with antiproliferative, antitumour, antiviral and immunmodulatory effect; it signals through the Jak-STAT signal transduction pathway where signal transducer and activator of transcription 1 plays an important role.Preliminary experiments on the identification of the molecules that reinducing signal transducers and activators of transcription 1 indicate that interferon-gamma may be the responsible candidate cytokine, but several others may be involved as well.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology and Oncology, Medizinische Hochschule, Hannover, Germany.

Show MeSH
Related in: MedlinePlus