title: Real-time monitoring of miRNA function in pancreatic cell lines using recombinant AAV-based miRNA Asensors.
abstract: BACKGROUND
  MicroRNAs (miRNAs) are reportedly involved in pancreatic ductal adenocarcinoma (PDAC) development. Current methods do not allow us to reliably monitor miRNA function. Asensors are adeno-associated virus (AAV) vector miRNA sensors for real-time consecutive functional monitoring of miRNA profiling in living cells.
  
  
  METHODS
  miR-200a, -200b, -21, -96, -146a, -10a, -155, and -221 in three PDAC cell lines (BxPC-3, CFPAC-1, SW1990), pancreatic epithelioid carcinoma cells (PANC-1), and human pancreatic nestin-expressing cells (hTERT-HPNE) were monitored by Asensors. Subsequently, the real-time consecutive functional profile of all miRNAs was evaluated.
  
  
  RESULTS
  Selected miRNAs were detectable in all cell lines with high sensitivity and reproducibility. In the three PDAC cell lines, BxPC-3, CFPAC-1, and SW1990, the calibrated signal unit of the eight miRNAs Asensors was significantly lower than that of the Asensor control. However, in PANC-1 cells, miR-200a and -155 showed upregulation of target gene expression at 24 hours after infection with the sensors; at 48 hours, miR-200b and -155 displayed upregulation of reporter expression; and at 72 hours, reporter gene expression was upregulated by miR-200a and -200b. The result that miRNA could upregulate gene expression was further confirmed in miR-155 of hTERT-HPNE cells. Furthermore, miRNA activity varied among cell/tissue types and time.
  
  
  CONCLUSIONS
  It is possible that miRNA participates in the pathophysiology of pancreatic cancer, but the current popular methods do not accurately reveal the real-time miRNA function. Thus, this report provided a convenient, accurate, and sensitive approach to miRNA research.
authors.affiliation: Department of Gastroenterology, Beijing Chaoyang Hospital, Capital Medical University, Chaoyang District, Beijing, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Chen
authors.forename: Xinjuan
authors.initials: X
authors.lastname: Liu
authors.forename: Xue
authors.initials: X
authors.lastname: Chen
authors.forename: Zihao
authors.initials: Z
authors.lastname: Guo
authors.forename: Juan
authors.initials: J
authors.lastname: Liu
authors.forename: Jianyu
authors.initials: J
authors.lastname: Hao
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
mesh_terms: D002460:Cell Line; D000229:Dependovirus / Q000235:genetics*; D006801:Humans; D035683:MicroRNAs / Q000235:genetics*; D010179:Pancreas / Q000378:metabolism*; D010190:Pancreatic Neoplasms
chemical_list: D035683:MicroRNAs