title: hCLCA1 DNA vaccine suppresses cell hyperplasia and mucin expression of goblet cells in vitro.
abstract: BACKGROUND
  Excessive airway mucus secretion is a remarkable trait of asthma. Mucus overproduction mainly resulted from an increase in goblet cell numbers, which causes considerable damage to health. However, effective therapeutic treatments are still lacking for mucus hypersecretion. Human calcium-activated chloride channel 1 (hCLCA1) has been identified to be predominantly responsible for mucus hypersecretion.
  
  
  OBJECTIVE
  In this study, we investigated the effects of an hCLCA1 DNA vaccine on the control of mucus production and goblet cell proliferation using an in vitro model goblet cell line (NCI-H292).
  
  
  METHODS
  The effect of the hCLCA1 DNA vaccine on cell viability and proliferative activity of NCI-H292/hCLCA1 was analyzed by electron microscopy, MTT assay, and flow cytometry. Expression of mucins and MUC5AC, a major member of the mucin gene family in airway goblet cells, was assessed under hCLCA1 DNA vaccine challenges by periodic acid-Schiff staining, quantitative real-time PCR and Western blot, respectively, and the expression profile of granulocyte-macrophage colony-stimulating factor (GM-CSF), a critical cytokine in airway inflammation, was also examined by real-time PCR and immunocytochemistry.
  
  
  RESULTS
  Results showed that hCLCA1 overexpression caused high cell proliferation and mucin expression, whereas the hCLCA1 DNA vaccine could effectively reverse these abnormal effects. In addition, GM-CSF expression was highly induced by hCLCA1 overexpression and efficiently suppressed by hCLCA1 DNA vaccine.
  
  
  CONCLUSIONS
  These results illustrate that the hCLCA1 DNA vaccine effectively inhibits cell hyperplasia and mucin gene expression of goblet cells, suggesting that the hCLCA1 DNA vaccine has potential value in the treatment of human asthma.
authors.affiliation: Department of Respiratory Medicine, Xijing Hospital, Xi'an, China.
authors.forename: L Q
authors.initials: LQ
authors.lastname: Song
authors.forename: Y
authors.initials: Y
authors.lastname: Li
authors.forename: W N
authors.initials: WN
authors.lastname: Li
authors.forename: W
authors.initials: W
authors.lastname: Zhang
authors.forename: Y Q
authors.initials: YQ
authors.lastname: Yang
authors.forename: H W
authors.initials: HW
authors.lastname: Qi
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D049109:Cell Proliferation; D002470:Cell Survival; D018118:Chloride Channels / Q000235:genetics*; D015536:Down-Regulation; D005434:Flow Cytometry; D020397:Goblet Cells / Q000276:immunology / Q000473:pathology*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics / Q000276:immunology; D006801:Humans; D006965:Hyperplasia; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron; D055271:Mucin 5AC / Q000235:genetics / Q000276:immunology*; D009077:Mucins / Q000235:genetics / Q000276:immunology*; D012333:RNA, Messenger / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D019444:Vaccines, DNA / Q000494:pharmacology*
chemical_list: D018118:Chloride Channels; C524678:MUC5AC protein, human; D055271:Mucin 5AC; D009077:Mucins; D012333:RNA, Messenger; D019444:Vaccines, DNA; D016178:Granulocyte-Macrophage Colony-Stimulating Factor