title: Regulation of connexin 43 and microRNA expression via β2-adrenoceptor signaling in 1321N1 astrocytoma cells.
abstract: Connexin 43 (Cx43) is the main gap junction protein in astrocytes and exerts the same effects on growth inhibition in astrocytoma and glioma as microRNA-146a (miR-146a) in glioma. β2-adrenergic receptor (AR) signaling modulates Cx43 expression in myocytes via components downstream of protein kinase A (PKA) and exchange protein directly activated by cAMP (Epac). However, it remains to be elucidated how expression of Cx43 is modulated in astrocytes. In the present study, 1321N1 astrocytoma cells were treated with β2-AR signaling agents in order to evaluate the expression of Cx43 and miRNAs. RNA and protein were extracted from the cells for use in reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. The results revealed that clenbuterol increased miR-146a level and upregulated Cx43 expression via cAMP/PKA at the mRNA and protein level. Pre-inhibition of adenyl cyclase decreased expression of Cx43 and miR-146a. PKA activation and overexpression of miR-146a in A-1321N1 cells increased the expression of Cx43. β2-AR stimulation and 6Bnz, a PKA activator, suppressed oncomiRs miR-155 and miR-27a, while 8-(4-chlorophenylthio)-2'-O-methyladenosine-3',5'-cyclic monophosphate, an Epac activator, increased their levels. The current findings demonstrated that β2-AR signaling has growth inhibitory effects via modulation of the cAMP/PKA pathway in A-1321N1 cells through increasing the expression level of Cx43 and miR-146a as well as decreasing miR-155 and miR-27a levels. Thus, stimulation of the β2-AR and PKA signaling pathway may be a useful approach for astrocytoma therapy.
authors: Department of Physiology, School of Medicine, Lorestan University of Medical Sciences, Khorramabad 6814993165, Iran.
authors: Mojtaba
authors: M
authors: Khaksarian
authors: Department of Molecular Biology and Genetic Engineering, Stem Cell Technology Research Center, Tehran 1997775555, Iran.
authors: Hossein
authors: H
authors: Mostafavi
authors: Department of Molecular Biology and Genetic Engineering, Stem Cell Technology Research Center, Tehran 1997775555, Iran.
authors: Masoud
authors: M
authors: Soleimani
authors: Department of Physiology, School of Medicine, Tehran University of Medical Sciences, Tehran 1417613151, Iran.
authors: Seid Morteza
authors: SM
authors: Karimian
authors: Department of Molecular Medicine, School of Advanced Medical Technologies, Tehran University of Medical Sciences, Tehran 1417743371, Iran.
authors: Mohammad Hassan
authors: MH
authors: Ghahremani
authors: Department of Anatomy, School of Medicine, Iran University of Medical Sciences, Tehran 1417755469, Iran.
authors: Mohammad Taghee
authors: MT
authors: Joghataee
authors: Department of Medical Biotechnologies, School of Advanced Medical Technologies, Tehran University of Medical Sciences, Tehran 1417743371, Iran.
authors: Mohsen
authors: M
authors: Khorashadizadeh
authors: Department of Neuroscience, School of Advanced Medical Technologies, Tehran University of Medical Sciences, Tehran 1417743371, Iran.
authors: Hadi
authors: H
authors: Aligholi
authors: Department of Neuroscience, School of Advanced Medical Technologies, Tehran University of Medical Sciences, Tehran 1417743371, Iran.
authors: Fatemeh
authors: F
authors: Attari
authors: Department of Neuroscience, School of Advanced Medical Technologies, Tehran University of Medical Sciences, Tehran 1417743371, Iran.
authors: Gholamreza
authors: G
authors: Hassanzadeh
mesh_terms: D001254:Astrocytoma / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D018031:Connexin 43 / Q000235:genetics*; D000242:Cyclic AMP / Q000378:metabolism; D017362:Cyclic AMP Response Element-Binding Protein / Q000235:genetics; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D057809:HEK293 Cells; D006801:Humans; D035683:MicroRNAs / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D018343:Receptors, Adrenergic, beta-2 / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: C115163:CREB1 protein, human; D018031:Connexin 43; D017362:Cyclic AMP Response Element-Binding Protein; C500216:GJA1 protein, human; D035683:MicroRNAs; D012333:RNA, Messenger; D018343:Receptors, Adrenergic, beta-2; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases