mRNA Relative Expression of Estrogen Receptors in Peripheral Blood Mononuclear Cells (PBMC) of Healthy Donors

CARDOZO MA; GAYDOU L; FOLLONIER A; BOSQUIAZZO VL; RAMOS JG

Congreso; Reunión Conjunta de Sociedades de Biociencias I; 2017

SAIC

Cell free DNA (cfDNA) in plasma has the clinical potential to be a more specific tumour marker fordiagnosis and prognosis, as well as for early disease detection. Currently, determination of theJAK2V617F mutation and measurement of its allele burden (AB) is performed in genomic DNA(gDNA) using both peripheral blood (PB) and bone marrow (BM) samples from patients withchronic myeloproliferative neoplasms (MPNs). With this procedure, not all malignant alleles couldbe detected. In this work we compared the %AB of JAK2V617F (percentage of allelesJAK2V617F / total alleles JAK2) in gDNA and cfDNA samples using a quantitative rel time PCRmethodology. For quantification we used a calibration curve made with a mixture of gDNA fromhealthy individuals and gDNA derived from a homozygous JAK2V617F-Human-erythroleukemiacell line (ATCC Hel 92.1.7). JAK2 status mutation was analyzed in 10 patients with MPNs, primaryand secondary myelofibrosis (MF) [n: 4], Escential thrombocytosis (ET) and Policitema Vera (PV)(n: 6), which were diagnosed according to the 2008 World Health Organization (WHO)classification. gDNA was obtained from peripheral blood using a modified Miller and Dykestechnique, while cfDNA was obtained from plasma using the QIAmp DNA Blood Mini kit. Weshowed a highly significative correlation between the %AB of JAK2V617F in cfDNA and gDNA(Spearman P=0.0028). Also, when we analyzed paired samples, we observed a higher %AB incfDNA (P=0.001) compared to the gDNA compartment. These data show that plasma is enrichedwith tumour-specific nucleic acid and could be the sample of choice for testing JAK2 mutations.