Downregulation of aquaporin 5 induced by vector-based short hairpin RNA and its effect on MUC5AC gene expression in human airway submucosal gland cells.
The aquaporins (AQPs) are a family of homologous water channels expressed in many epithelial and endothelial cells, however no reliable and non-toxic inhibitors of AQPs have been reported yet. Our researchers have analyzed the changes of AQP5 expression induced by vector-based short hairpin RNA (shRNA) in the human airway submucosal gland cell line (SPC-A1) and observed its regulation on the expression of MUC5AC gene. Localizations of AQP5 and MUC5AC in SPC-A1cells were detected by Immunofluorescence. AQP5 mRNA was significantly reduced by 75.1% one day after transfection with specific shRNA, named shAQP5. However, the significant suppression of AQP5 protein did not appear until day 5 after transfection. MUC5AC mRNA was remarkably increased by 119.9% On day 3 after shAQP5 transfection, while comparable MUC5AC protein changes were not found in SPC-A1 cells with flow cytometry analysis. These results indicate that vector-based shRNA could be used as a potential tool to inhibit the expression of AQP5. This is the first investigation providing evidence demonstrating the regulation of the mucin gene by AQP5 gene silencing.
Chen
Zhihong
Z
Department of Pulmonary Medicine, Research Institute of Respiratory Disease, Fudan University, Zhongshan Hospital, Shanghai, PR China.
Zhu
Rong
R
Bai
Li
L
Bai
Chunxue
C
D000704:Analysis of Variance; D051402:Aquaporin 5 / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western / Q000379:methods; D002277:Carcinoma; D045744:Cell Line, Tumor; D015536:Down-Regulation / Q000187:drug effects*; D005434:Flow Cytometry / Q000379:methods; D005455:Fluorescent Antibody Technique / Q000379:methods; D015870:Gene Expression / Q000502:physiology*; D005822:Genetic Vectors / Q000502:physiology; D049452:Green Fluorescent Proteins / Q000096:biosynthesis; D006801:Humans; D008175:Lung Neoplasms; D055271:Mucin 5AC; D009077:Mucins / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013997:Time Factors; D014162:Transfection
D051402:Aquaporin 5; C524678:MUC5AC protein, human; D055271:Mucin 5AC; D009077:Mucins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins