title: miR-140-3p regulation of TNF-α-induced CD38 expression in human airway smooth muscle cells.
abstract: CD38, a membrane protein expressed in airway smooth muscle (ASM) cells, plays a role in cellular Ca(2+) dynamics and ASM contractility. In human ASM (HASM) cells, TNF-α induces CD38 expression through activation of MAPKs, NF-κB, and AP-1, and its expression is differentially elevated in cells from asthmatic patients compared with cells from nonasthmatic subjects. The CD38 3'-untranslated region (UTR) has targets for miR-140-3p. We hypothesized that miR-140-3p regulates CD38 expression in HASM cells by altering CD38 mRNA stability. Basal and TNF-α-induced expression of miR-140-3p was determined in nonasthmatic ASM (NAASM) and asthmatic ASM (AASM) cells. NAASM and AASM cells were transfected with control, miR-140-3p mimic, or miR-140-3p antagomirs, and CD38 expression and CD38 mRNA stability were determined. Luciferase reporter assays were used to determine miR-140-3p binding to the CD38 3'-UTR. Activation of p38, ERK, and JNK MAPKs, NF-κB, and AP-1 was determined in miR-140-3p mimic-transfected NAASM. TNF-α attenuated miR-140-3p expression in NAASM and AASM cells, but at a greater magnitude in AASM cells. CD38 mRNA expression was attenuated by miR-140-3p mimic at comparable magnitude in NAASM and AASM cells. Mutated miR-140-3p target on the CD38 3'-UTR reversed the inhibition of luciferase activity by miR-140-3p mimic. CD38 mRNA stability was unaltered by miR-140-3p mimic in NAASM or AASM cells following arrest of transcription. TNF-α-induced activation of p38 MAPK and NF-κB was attenuated by miR-140-3p mimic. The findings indicate that miR-140-3p modulates CD38 expression in HASM cells through direct binding to the CD38 3'-UTR and indirect mechanisms involving activation of p38 MAPK and NF-κB. Furthermore, indirect mechanisms appear to play a major role in the regulation of CD38 expression.
authors: Dept. of Veterinary and Biomedical Sciences, College of Veterinary Medicine, Univ. of Minnesota, St. Paul, MN 55108, USA.
authors: Joseph A
authors: JA
authors: Jude
authors: Mythili
authors: M
authors: Dileepan
authors: Subbaya
authors: S
authors: Subramanian
authors: Julian
authors: J
authors: Solway
authors: Reynold A
authors: RA
authors: Panettieri
authors: Timothy F
authors: TF
authors: Walseth
authors: Mathur S
authors: MS
authors: Kannan
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics; D051997:ADP-ribosyl Cyclase 1 / Q000235:genetics / Q000378:metabolism*; D001249:Asthma / Q000378:metabolism / Q000473:pathology; D002478:Cells, Cultured; D015536:Down-Regulation; D015870:Gene Expression; D006801:Humans; D020935:MAP Kinase Signaling System; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism; D034622:RNA Interference; D012137:Respiratory System / Q000378:metabolism / Q000473:pathology*; D014409:Tumor Necrosis Factor-alpha / Q000502:physiology*; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
chemical_list: D020413:3' Untranslated Regions; D008562:Membrane Glycoproteins; D035683:MicroRNAs; C544302:Mirn140 microRNA, human; D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha; D048051:p38 Mitogen-Activated Protein Kinases; C498154:CD38 protein, human; D051997:ADP-ribosyl Cyclase 1