Quantitative detection of methylated ESR1 and 14-3-3-sigma gene promoters in serum as candidate biomarkers for diagnosis of breast cancer and evaluation of treatment efficacy.
The aim of the present study was to investigate the association between gene hypermethylation and main clinicopathological features of breast cancer, including diagnosis and treatment response. A sensitive SYBR green methylation-specific PCR technique was used to analyze the utility of circulating DNA with CpG island hypermethylation of ESR1, APC, RARB, 14-3-3-sigma and E-cad gene promoter regions as breast cancer biomarkers. Analyses were conducted of preoperative sera from 106 women with breast cancer, 34 with benign breast disease and 74 with no evidence of breast disease and of post-treatment sera from 60 of the breast cancer patients. Mean serum values of methylated ESR1 and 14-3-3-sigma gene promoters significantly differed between breast cancer patients and healthy controls (p = 0.0112 for ESR1 and p = 0.0047 for 14-3-3-sigma). When their results were combined, it was found that hypermethylation of these two genes differentiated between breast cancer patients and healthy controls (p < 0.0001) with a sensitivity of 81% (95% confidence interval: 72-88%) and specificity of 88% (95% CI: 78-94%). Presence of methylated ESR1 in serum of breast cancer patients was associated with the ER negative phenotype (p = 0.0179). Serum hypermethylation at ESR1 and 14-3-3-sigma loci was observed in cancer patients, in situ carcinoma and benign breast disease. No significant differences in methylated ERS1 or 14-3-3-sigma values were observed between pre-surgery and post-treatment measurements. Preliminary clinical applications of this approach have revealed several shortcomings, including a frequent presence of methylated 14-3-3-sigma in sera from women with breast benign disease. These findings cast some doubts on the utility for early cancer diagnosis of highly sensitive techniques to identify hypermethylation of specific gene promoters in DNA extracted from serum. Although numerous issues remain to be resolved, the quantitative measurement of circulating methylated DNA remains a promising tool for cancer risk assessment.
Martínez-Galán
Joaquina
J
Servicio de Oncología Radioterápica, Hospital Virgen de las Nieves, Granada, Spain.
Torres
Blanca
B
Del Moral
Rosario
R
Muñoz-Gámez
José Antonio
JA
Martín-Oliva
David
D
Villalobos
Mercedes
M
Núñez
María Isabel
MI
Luna
Juan de Dios
Jde D
Oliver
Francisco Javier
FJ
Ruiz de Almodóvar
José Mariano
JM
D048948:14-3-3 Proteins; D014408:Biomarkers, Tumor / Q000378:metabolism*; D001943:Breast Neoplasms / Q000097:blood* / Q000175:diagnosis*; D016022:Case-Control Studies; D019175:DNA Methylation; D004273:DNA, Neoplasm / Q000378:metabolism; D018450:Disease Progression; D047628:Estrogen Receptor alpha / Q000378:metabolism*; D005092:Exonucleases / Q000378:metabolism*; D005095:Exoribonucleases; D005260:Female; D006801:Humans; D009363:Neoplasm Proteins / Q000378:metabolism*; D010641:Phenotype; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012680:Sensitivity and Specificity; D016896:Treatment Outcome
D048948:14-3-3 Proteins; D014408:Biomarkers, Tumor; D004273:DNA, Neoplasm; D047628:Estrogen Receptor alpha; D009363:Neoplasm Proteins; D005092:Exonucleases; D005095:Exoribonucleases; C413442:SFN protein, human