title: Atrial natriuretic peptide: A novel mediator for TGF-β1-induced epithelial-mesenchymal transition in 16HBE-14o and A549 cells.
abstract: Atrial natriuretic peptide (ANP) is increasingly expressed on airway and inhibits pulmonary arterial remodeling. However, the role of ANP in remodeling of respiratory system is still unclear. The role of ANP on airway remodeling and the possible mechanism was explored in this study. Both human bronchial epithelial 16HBE-14o cells and alveolar epithelial A549 cells were stimulated by TGF-β1, ANP, cGMP inhibitor, PKG inhibitor, and cGMP analogue. The expressions of epithelial markers, mesenchymal markers, and Smad3 were assessed by quantitative real-time PCR and western blotting. Immunohistochemical staining was employed to assess Smad3 expression once it was silenced by siRNA in 16HBE-14o or A549 cells. Our results showed that the mRNA and protein expressions of E-Cadherin were decreased, whereas α-SMA expressions were increased after induction by TGF-β1 in 16HBE-14o and A549 cells. The E-Cadherin expressions were increased and α-SMA expressions were decreased after ANP stimulation. Inhibition of cGMP or PKG decreased E-Cadherin expression but increased α-SMA expression, which could be reversed by cGMP analogue. Moreover, the phosphorylated Smad3 expression was consistent with α-SMA expression. After smad3 was silenced, Smad3 was mostly expressed in cytoplasm instead of nucleus as non-silenced cells during epithelial-mesenchymal transition (EMT). In conclusion, ANP inhibits TGF-β1-induced EMT in 16HBE-14o and A549 cells through cGMP/PKG signaling, by which it targets TGF-β1/Smad3 via attenuating phosphorylation of Smad3. These findings suggest the potential of ANP in the treatment on pulmonary diseases with airway remodeling.
authors.affiliation: Department of Respiratory Medicine, The Affiliated Hospital of Guilin Medical University, Guilin 541001, Guangxi, China; Institute of Respiratory Diseases, Guilin Medical University, Guilin 541001, Guangxi, China; Guangxi Colleges and Universities Key Laboratory of Respiratory Disease, Guilin 541001, Guangxi, China.
authors.forename: Shuyuan
authors.initials: S
authors.lastname: Chu
authors.affiliation: Department of Respiratory Medicine, The Second Affiliated Hospital of University of South China, Hengyang 421000, Hunan, China.
authors.forename: Xiufeng
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Respiratory Medicine, The Affiliated Hospital of Guilin Medical University, Guilin 541001, Guangxi, China; Institute of Respiratory Diseases, Guilin Medical University, Guilin 541001, Guangxi, China; Guangxi Colleges and Universities Key Laboratory of Respiratory Disease, Guilin 541001, Guangxi, China.
authors.forename: Yabing
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Department of Respiratory Medicine, The Affiliated Hospital of Guilin Medical University, Guilin 541001, Guangxi, China; Institute of Respiratory Diseases, Guilin Medical University, Guilin 541001, Guangxi, China; Guangxi Colleges and Universities Key Laboratory of Respiratory Disease, Guilin 541001, Guangxi, China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Yu
authors.affiliation: Department of Respiratory Medicine, The Affiliated Hospital of Guilin Medical University, Guilin 541001, Guangxi, China; Institute of Respiratory Diseases, Guilin Medical University, Guilin 541001, Guangxi, China; Guangxi Colleges and Universities Key Laboratory of Respiratory Disease, Guilin 541001, Guangxi, China.
authors.forename: Yaxi
authors.initials: Y
authors.lastname: Liang
authors.affiliation: Department of Respiratory Medicine, The Affiliated Hospital of Guilin Medical University, Guilin 541001, Guangxi, China; Institute of Respiratory Diseases, Guilin Medical University, Guilin 541001, Guangxi, China; Guangxi Colleges and Universities Key Laboratory of Respiratory Disease, Guilin 541001, Guangxi, China.
authors.forename: Ming
authors.initials: M
authors.lastname: Jiang
authors.affiliation: Department of Respiratory Medicine, The Affiliated Hospital of Guilin Medical University, Guilin 541001, Guangxi, China; Institute of Respiratory Diseases, Guilin Medical University, Guilin 541001, Guangxi, China; Guangxi Colleges and Universities Key Laboratory of Respiratory Disease, Guilin 541001, Guangxi, China.
authors.forename: Jianwei
authors.initials: J
authors.lastname: Huang
authors.affiliation: Department of Respiratory Medicine, The Affiliated Hospital of Guilin Medical University, Guilin 541001, Guangxi, China; Institute of Respiratory Diseases, Guilin Medical University, Guilin 541001, Guangxi, China; Guangxi Colleges and Universities Key Laboratory of Respiratory Disease, Guilin 541001, Guangxi, China. Electronic address: malibing1984@163.com.
authors.forename: Libing
authors.initials: L
authors.lastname: Ma
mesh_terms: D000072283:A549 Cells; D000199:Actins / Q000235:genetics / Q000378:metabolism; D056151:Airway Remodeling / Q000235:genetics; D009320:Atrial Natriuretic Factor / Q000235:genetics* / Q000378:metabolism; D015820:Cadherins / Q000235:genetics*; D006152:Cyclic GMP / Q000037:antagonists & inhibitors / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics; D005786:Gene Expression Regulation / Q000235:genetics; D006801:Humans; D008171:Lung Diseases / Q000188:drug therapy / Q000235:genetics / Q000473:pathology; D015398:Signal Transduction / Q000187:drug effects; D051900:Smad3 Protein / Q000235:genetics*; D053773:Transforming Growth Factor beta1 / Q000008:administration & dosage / Q000378:metabolism*
chemical_list: C541116:ACTA2 protein, human; D000199:Actins; D015820:Cadherins; C497790:SMAD3 protein, human; D051900:Smad3 Protein; D053773:Transforming Growth Factor beta1; D009320:Atrial Natriuretic Factor; D006152:Cyclic GMP
keywords: Alveolar epithelial cells; Atrial natriuretic peptide; Bronchial epithelial cells; Epithelial-mesenchymal transition