Macrophage colony-stimulating factor (M-CSF) is a multifunctional cytokine attributed with key biological functions beyond the first discovered role in promoting proliferation of myeloid cell lineage. The human pancreatic cancer cell line MIA PaCa-2, from which the M-CSF gene was originally cloned, was used to study regulation of M-CSF expression. Expression of M-CSF was inducible by interleukin-1α (IL-1α), lipopolysaccharide (LPS) and PMA as demonstrated by a biological activity assay, Northern-blot analysis and reverse transcriptase (RT) PCR. Treatment of the cells with forskolin or dibutyryl-cAMP attenuated the expression of M-CSF induced by IL-1α or LPS, but not by PMA. Electromobility shift assays showed that IL-1α predominantly activated nuclear factor κB (NF-κB), while PMA preferentially activated activator protein-1 (AP-1). The activation of NF-κB, but not AP-1, could be attenuated by cAMP elevation. Relative RT-PCR demonstrated that the expression of a 1.6-kb M-CSF mRNA transcript was more effectively induced by IL-1α than a 4.0-kb transcript. By and large the induced expression of both mRNA transcripts could be attenuated by cAMP. M-CSF promoter-driven luciferase reporter-gene assays revealed that cAMP elevation attenuated the IL-1-induced transcription activation of the M-CSF promoter, but it had no effect on PMA-induced transcription. Our findings suggest that cAMP regulates M-CSF gene expression at the transcriptional level and that its inhibitory effect involves NF-κB signalling pathway.
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Research Article| August 09 2000
cAMP attenuates interleukin-1-stimulated macrophage colony-stimulating factor (M-CSF) expression
Pisate J. KAMTHONG ;
Fu-mei WU ;
Ming-chi WU 1
1Department of Molecular Biology and Immunology, University of North Texas Health Science Center at Fort Worth, 3500 Camp Bowie Blvd, Fort Worth, TX 76107, U.S.A.
1To whom correspondence should be addressed (e-mail firstname.lastname@example.org).
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Pisate J. KAMTHONG, Fu-mei WU, Ming-chi WU; cAMP attenuates interleukin-1-stimulated macrophage colony-stimulating factor (M-CSF) expression. Biochem J 15 August 2000; 350 (1): 115–122. doi: https://doi.org/10.1042/bj3500115
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