title: Repeated Aurora-A siRNA Transfection Results in Effective Apoptosis of A549 Cells Compared to Single Transfection.
abstract: BACKGROUND
  Suppression of Aurora kinase A (Aurora-A, AURKA) by Aurora-A siRNA has been proposed for lung tumor treatment. However, protocols using single administration have shown little benefit in some types of lung tumor. Given that transfection efficiency of Aurora-A siRNA is low due to tightly packed cells in the tumor, we hypothesized that repeated administration would result in efficient cell apoptosis.
  
  
  METHODS
  We compared single vs. repeated transfection (thrice) in A549 cells by transfecting Aurora-A siRNA (siA) on the 1st or 1st, 2nd and 3rd day after cell seeding. A random sequence was used as the negative siRNA control (siC). Cells in the single transfection group received only transfection reagent without siRNAs on the 2nd and 3rd day.
  
  
  RESULTS
  Two days after the third transfection, both single and repeated siA administration decreased mRNA expression of Aurora-A and cell viability compared to no administration and siC single administration. However, the decrease in these two indices with repeated transfection was more obvious than that following single administration: cell viability decreased to 72.8 ± 3.05% (p < 0.05) following siA single transfection and to 64.2 ± 1.99% (p < 0.05) following siA repeated transfection, compared with normal control cells, respectively. Gene expression decreased to 17 ± 16.6% (p < 0.05 vs. normal control) following siA repeated transfection and to 43.2 ± 13.0% (p < 0.05 vs. normal control) following siA single transfection.
  
  
  CONCLUSIONS
  Compared to single transfection, repeated Aurora-A siRNA transfection decreased Aurora-A, which, in turn, resulted in effective apoptosis of A549 cells.
authors.forename: Zhonghua
authors.initials: Z
authors.lastname: Wang
authors.forename: Wenwu
authors.initials: W
authors.lastname: Sun
authors.forename: Jianping
authors.initials: J
authors.lastname: Cao
authors.forename: Haiyang
authors.initials: H
authors.lastname: Cui
authors.forename: Zhuang
authors.initials: Z
authors.lastname: Ma
mesh_terms: D017209:Apoptosis; D064096:Aurora Kinase A / Q000235:genetics* / Q000502:physiology; D045744:Cell Line, Tumor; D002470:Cell Survival; D006801:Humans; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection
chemical_list: D034741:RNA, Small Interfering; C576917:AURKA protein, human; D064096:Aurora Kinase A