title: Epigenetic alteration: new insights moving from tissue to plasma - the example of PCDH10 promoter methylation in colorectal cancer.
abstract: BACKGROUND
  Tumour-released DNA in blood represents a promising biomarker for cancer detection. Although epigenetic alterations such as aberrant promoter methylation represent an appealing perspective, the discordance existing between frequencies of alterations found in DNA extracted from tumour tissue and cell-free DNA (cfDNA) has challenged their practical clinical application. With the aim to explain this bias of agreement, we investigated whether protocadherin 10 (PCDH10) promoter methylation in tissue was associated with methylation pattern in matched cfDNA isolated from plasma of patients with colorectal cancer (CRC), and whether the strength of concordance may depend on levels of cfDNA, integrity index, as well as on different clinical-pathological features.
  
  
  METHODS
  A quantitative methylation-specific PCR was used to analyse a selected CpG site in the PCDH10 promoter of 67 tumour tissues, paired normal mucosae, and matched plasma samples. The cfDNA integrity index and cfDNA concentration were assessed using a real-time PCR assay.
  
  
  RESULTS
  The PCDH10 promoter methylation was detected in 63 out of 67 (94.0%) surgically resected colorectal tumours and in 42 out of 67 (62.7%) plasma samples. The median methylation rate in tumour tissues and plasma samples was 43.5% (6.3-97.8%) and 5.9% (0-80.9%), respectively. There was a significant correlation between PCDH10 methylation in cfDNA and tumour tissue in patients with early CRC (P<0.0001). The ratio between plasma and tissue methylation rate increases with increasing cfDNA integrity index in early-stage cancers (P=0.0299) and with absolute cfDNA concentration in advanced cancers (P=0.0234).
  
  
  CONCLUSIONS
  Our findings provide new insight into biological aspects modulating the concordance between tissues and plasma methylation profiles.
authors.affiliation: Department of Medicine, University Hospital of Verona, Hospital Policlinico G.B. Rossi, Verona, Italy. elisa.danese@univr.it
authors.forename: E
authors.initials: E
authors.lastname: Danese
authors.forename: A M
authors.initials: AM
authors.lastname: Minicozzi
authors.forename: M
authors.initials: M
authors.lastname: Benati
authors.forename: M
authors.initials: M
authors.lastname: Montagnana
authors.forename: E
authors.initials: E
authors.lastname: Paviati
authors.forename: G L
authors.initials: GL
authors.lastname: Salvagno
authors.forename: M
authors.initials: M
authors.lastname: Gusella
authors.forename: F
authors.initials: F
authors.lastname: Pasini
authors.forename: G C
authors.initials: GC
authors.lastname: Guidi
authors.forename: G
authors.initials: G
authors.lastname: Lippi
mesh_terms: D015820:Cadherins / Q000235:genetics*; D015331:Cohort Studies; D015179:Colorectal Neoplasms / Q000097:blood* / Q000235:genetics* / Q000473:pathology; D019175:DNA Methylation; D004273:DNA, Neoplasm / Q000097:blood / Q000235:genetics* / Q000302:isolation & purification; D015536:Down-Regulation; D020868:Gene Silencing; D006801:Humans; D011401:Promoter Regions, Genetic; D000091363:Protocadherins
chemical_list: D015820:Cadherins; D004273:DNA, Neoplasm; C438603:PCDH10 protein, human; D000091363:Protocadherins