title: Automated cell disruption is a reliable and effective method of isolating RNA from fresh snap-frozen normal and malignant oral mucosa samples.
abstract: BACKGROUND
  This study compared automated vs. manual tissue grinding in terms of RNA yield obtained from oral mucosa biopsies.
  
  
  METHODS
  A total of 20 patients undergoing uvulectomy for sleep-related disorders and 10 patients undergoing biopsy for head and neck squamous cell carcinoma were enrolled in the study. Samples were collected, snap-frozen in liquid nitrogen, and divided into two parts of similar weight. Sample grinding was performed on one sample from each pair, either manually or using an automated cell disruptor. The performance and efficacy of each homogenization approach was compared in terms of total RNA yield (spectrophotometry, fluorometry), mRNA quantity [densitometry of specific TP53 amplicons and TP53 quantitative reverse-transcribed real-time PCR (qRT-PCR)], and mRNA quality (functional analysis of separated alleles in yeast).
  
  
  RESULTS
  Although spectrophotometry and fluorometry results were comparable for both homogenization methods, TP53 expression values obtained by amplicon densitometry and qRT-PCR were significantly and consistently better after automated homogenization (p<0.005) for both uvula and tumor samples. Functional analysis of separated alleles in yeast results was better with the automated technique for tumor samples.
  
  
  CONCLUSIONS
  Automated tissue homogenization appears to be a versatile, quick, and reliable method of cell disruption and is especially useful in the case of small malignant samples, which show unreliable results when processed by manual homogenization.
authors.affiliation: Center for Applied Molecular Technology, Université catholique de Louvain, Brussels, Belgium.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Van der Vorst
authors.forename: Anne-France
authors.initials: AF
authors.lastname: Dekairelle
authors.forename: Léonid
authors.initials: L
authors.lastname: Irenge
authors.forename: Marc
authors.initials: M
authors.lastname: Hamoir
authors.forename: Annie
authors.initials: A
authors.lastname: Robert
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Gala
mesh_terms: D006801:Humans; D009061:Mouth Mucosa / Q000473:pathology*; D009062:Mouth Neoplasms / Q000175:diagnosis*; D012313:RNA / Q000235:genetics / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013048:Specimen Handling / Q000379:methods; D013064:Speech Disorders / Q000601:surgery; D046509:Tissue Culture Techniques / Q000379:methods*; D016159:Tumor Suppressor Protein p53 / Q000235:genetics; D014609:Uvula / Q000601:surgery
chemical_list: D016159:Tumor Suppressor Protein p53; D012313:RNA