title: The effect of colonoscopy on whole blood gene expression profile: an experimental investigation for colorectal cancer biomarker discovery.
abstract: OBJECTIVE
  Gene expression profiling of whole blood is showing great promise for the discovery of novel biomarkers for colorectal cancer (CRC) detection. Given the relatively low incidence rate of CRC in the general population, most blood samples collected prior to a colonoscopy were confirmed to be noncancerous afterward. Previous studies have relied on blood samples collected after a colonoscopy to reach the sufficient number of CRC cases. The present study aimed to determine the colonoscopy-induced variability in the blood transcriptome and its potential impact on biomarker discovery.
  
  
  METHODS
  Whole blood gene expression profiling was conducted using Affymetrix HG-U133Plus2 arrays. We analyzed 20 paired blood samples collected from healthy controls before and after colonoscopy, and 20 blood samples collected from CRC patients after colonoscopy.
  
  
  RESULTS
  Utilizing hierarchical clustering analysis, samples collected before and after colonoscopy from the same subjects were closely clustered together, suggesting that the blood gene expression profiles primarily reflect the heterogeneity within each individual. A total of 914 genes were differentially expressed between controls and CRC patients, while gene expression did not differ between samples collected before and after colonoscopy. Using real-time PCR technology, we further validated six genes from published biomarkers in this study. Our results confirmed that the biomarkers identified in previous studies were not likely to be biased due to the colonoscopy effect.
  
  
  CONCLUSIONS
  Our results showed that the colonoscopy-induced variations were minute compared to individuals' heterogeneity and disease-induced variations. These findings may serve as a basis for future development of blood-based transcriptomic biomarkers for the diagnosis and treatment of CRC.
authors.affiliation: Department of Oncology, Fudan University Shanghai Medical College, Shanghai, China.
authors.forename: Ye
authors.initials: Y
authors.lastname: Xu
authors.forename: Qinghua
authors.initials: Q
authors.lastname: Xu
authors.forename: Li
authors.initials: L
authors.lastname: Yang
authors.forename: Fang
authors.initials: F
authors.lastname: Liu
authors.forename: Xun
authors.initials: X
authors.lastname: Ye
authors.forename: Fei
authors.initials: F
authors.lastname: Wu
authors.forename: Shujuan
authors.initials: S
authors.lastname: Ni
authors.forename: Cong
authors.initials: C
authors.lastname: Tan
authors.forename: Guoxiang
authors.initials: G
authors.lastname: Cai
authors.forename: Xia
authors.initials: X
authors.lastname: Meng
authors.forename: Sanjun
authors.initials: S
authors.lastname: Cai
authors.forename: Xiang
authors.initials: X
authors.lastname: Du
mesh_terms: D000328:Adult; D000368:Aged; D014408:Biomarkers, Tumor / Q000097:blood / Q000235:genetics*; D016000:Cluster Analysis; D003113:Colonoscopy; D015179:Colorectal Neoplasms / Q000097:blood / Q000235:genetics*; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods; D006801:Humans; D008297:Male; D008875:Middle Aged; D020411:Oligonucleotide Array Sequence Analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D059467:Transcriptome / Q000235:genetics*
chemical_list: D014408:Biomarkers, Tumor