title: A del T poly T (8) mutation in the 3' untranslated region (UTR) of the CDK2-AP1 gene is functionally significant causing decreased mRNA stability resulting in decreased CDK2-AP1 expression in human microsatellite unstable (MSI) colorectal cancer (CRC).
abstract: BACKGROUND
  We have previously published results indicating that decreased expression of CDK2-AP1 in MSI human colorectal cancer is associated with deletion mutations in the poly (T) 8 repeat sequence within the 3'-UTR of the CDK2-AP1 gene. In this study, we test the hypothesis that the del T mutation results in decreased CDK2-AP1 expression by causing reduced mRNA stability.
  
  
  METHODS
  We introduced wild-type and mutant 3'-UTR sequences fused to a green fluorescent protein (GFP) gene separately into human CRC cell lines and quantified the expression of the GFP gene. Native CDK2-AP1 mRNA stability was measured in human CRC cell lines, using an actinomycin D assay and the mRNA structure folding software mfold 3.2.
  
  
  RESULTS
  Mutant GFP-3'-UTR samples demonstrated significantly reduced GFP expression compared with wild-type GFP-3'-UTR as measured by both FACS and real-time PCR. Both the actinomycin D assay and mfold software demonstrated significantly reduced mRNA stability for the del T poly (T) 8 transcript compared with the wild type.
  
  
  CONCLUSIONS
  In summary, these novel results support our hypotheses that the del T poly (T) 8 observed in the 3'-UTR of the CDK2-AP1 gene in human MSI CRC is functionally significant and results in decreased CDK2-AP1 expression. The results also indicate the mechanism of this decreased expression is caused at least in part by decreased mRNA stability.
authors: Department of Surgery and Molecular Genetics, Albert Einstein College of Medicine, New York, NY, USA.
authors: Joongho
authors: J
authors: Shin
authors: Ziqian
authors: Z
authors: Yuan
authors: Kenneth
authors: K
authors: Fordyce
authors: Prashanth
authors: P
authors: Sreeramoju
authors: Tara S
authors: TS
authors: Kent
authors: John
authors: J
authors: Kim
authors: Victor
authors: V
authors: Wang
authors: David
authors: D
authors: Schneyer
authors: Thomas K
authors: TK
authors: Weber
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics; D045744:Cell Line, Tumor; D015179:Colorectal Neoplasms / Q000235:genetics* / Q000503:physiopathology; D017353:Gene Deletion; D015972:Gene Expression Regulation, Neoplastic; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D053842:Microsatellite Instability; D009690:Nucleic Acid Conformation; D020871:RNA Stability / Q000235:genetics*; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics; D012334:RNA, Neoplasm / Q000737:chemistry / Q000235:genetics; D025521:Tumor Suppressor Proteins / Q000235:genetics*
chemical_list: D020413:3' Untranslated Regions; C096151:CDK2AP1 protein, human; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D025521:Tumor Suppressor Proteins; D049452:Green Fluorescent Proteins