Evaluation of promoter hypermethylation detection in body fluids as a screening/diagnosis tool for head and neck squamous cell carcinoma.
OBJECTIVE
To evaluate aberrant promoter hypermethylation of candidate tumor suppressor genes as a means to detect epigenetic alterations specific to solid tumors, including head and neck squamous cell carcinoma (HNSCC).


METHODS
Using promoter regions identified via a candidate gene and discovery approach, we evaluated the ability of an expanded panel of CpG-rich promoters known to be differentially hypermethylated in HNSCC in detection of promoter hypermethylation in serum and salivary rinses associated with HNSCC. We did preliminary evaluation via quantitative methylation-specific PCR (Q-MSP) using a panel of 21 genes in a limited cohort of patients with HNSCC and normal controls. Using sensitivity and specificity for individual markers as criteria, we selected panels of eight and six genes, respectively, for use in salivary rinse and serum detection and tested these in an expanded cohort including up to 211 patients with HNSCC and 527 normal controls.


RESULTS
Marker panels in salivary rinses showed improved detection when compared with single markers, including a panel with 35% sensitivity and 90% specificity and a panel with 85% sensitivity and 30% specificity. A similar pattern was noted in serum panels, including a panel with 84.5% specificity with 50.0% sensitivity and a panel with sensitivity of 81.0% with specificity of 43.5%. We also noted that serum and salivary rinse compartments showed a differential pattern of methylation in normal subjects that influenced the utility of individual markers.


CONCLUSIONS
Q-MSP detection of HNSCC in serum and salivary rinses using multiple targets offers improved performance when compared with single markers. Compartment-specific methylation in normal subjects affects the utility of Q-MSP detection strategies.
Carvalho
André Lopes
AL
Department of Otolaryngology-Head and Neck Surgery, Johns Hopkins Medical Institutions, Baltimore, Maryland 21287-0910, USA.
Jeronimo
Carmen
C
Kim
Michael M
MM
Henrique
Rui
R
Zhang
Zhe
Z
Hoque
Mohammad O
MO
Chang
Steve
S
Brait
Mariana
M
Nayak
Chetan S
CS
Jiang
Wei-Wen
WW
Claybourne
Quia
Q
Tokumaru
Yutaka
Y
Lee
Juna
J
Goldenberg
David
D
Garrett-Mayer
Elizabeth
E
Goodman
Steven
S
Moon
Chul-so
CS
Koch
Wayne
W
Westra
William H
WH
Sidransky
David
D
Califano
Joseph A
JA
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001826:Body Fluids / Q000737:chemistry* / Q000378:metabolism; D002294:Carcinoma, Squamous Cell / Q000175:diagnosis* / Q000235:genetics; D019175:DNA Methylation; D004273:DNA, Neoplasm / Q000032:analysis; D005260:Female; D006258:Head and Neck Neoplasms / Q000175:diagnosis* / Q000235:genetics; D006801:Humans; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012463:Saliva / Q000737:chemistry / Q000378:metabolism; D012680:Sensitivity and Specificity; D044967:Serum / Q000737:chemistry / Q000378:metabolism
D004273:DNA, Neoplasm