DETERMINING SPECIFIC THYROID TRANSCRIPTS
IN PERIPHERAL BLOOD:
A SINGLE CENTER STUDY EXPERIENCE Makazlieva T, Eftimov A, Vaskova O, Tripunoski T,
Miladinova D, Risteski S, Jovanovic H, Jakovski Z
Tanja Makazlieva and Aleksandar Eftimov contributed equally to this study
and are considered first coauthors. *Corresponding Author: Tanja Makazlieva, Ph.D., Institute of Pathophysiology and Nuclear Medicine,
Medical Faculty, Mother Teresa Street, No. 17, 1000, Skopje, Republic of Macedonia.
Mobile: +389-75-313-665. E-mail: tmakazlieva@medf.ukim.edu.mk or tmakazlieva@gmail.com page: 13
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INTRODUCTION
Thyroid carcinoma (TC) comprises a spectrum of different
tumors with a wide range of biological behavior and
prognosis [1-3]. Differentiated thyroid carcinomas (DTC)
including papillary thyroid carcinoma (PTC) and follicular
thyroid carcinomas (FTC) originate from follicular epithelial
cells. The progression of DTC is usually slow, but
local involvement of lymph nodes is very often possible, as
well as loco-regional relapses as late as 30 years after the
initial diagnosis. In some cases, distant metastatic disease is
present even at initial presentation of the disease [4-6]. The
recommendations for long-term follow-up of DTC usually
include ultrasound of the neck, serum thyroglobulin (sTg)
levels and radioiodine whole body scan (131I WBS). The
sTg level is a very useful marker in detecting progression
of the disease, but this analysis has no diagnostic value in
patients with coexisting autoimmune thyroid disease and
positive anti-thyroglobulin antibodies (aTg). Endogenous
aTg may cause false-negative results in immunometric assays
for sTg determination, and sTg levels in those patients
may not reveal the exact state of the disease [7].
According to the new guidelines, in some patients with
microcarcinomas and low-risk tumors, the radioiodine ablation
(RAI) is not recommended, leading to a situation where
it is inconvenient to apply sTg as a tumor marker, imposing the need for new biomarkers of the disease [8]. Experiences
with diagnostic detection of circulating tumor cells in patients
with some solid tumors as breast, colon and prostate
carcinomas, are a motivation for researchers to apply similar
methodology in TC [9]. For this purpose, the techniques
based on the latest trends in molecular biology can have the
most appropriate application in diagnosis of metastatic TC.
The use of the real-time polymerase chain reaction
(RT-PCR) method for detection of circulating malignant
thyroid cells was first introduced by Ditkoff et al. [10]
evaluating Tg-mRNA transcripts in peripheral blood in both
healthy individuals and patients with TC, which yielded
promising results, leading to an increased number of published
papers on this topic in the years to come. The reports
about possible the usefulness of this approach in different
studies vary greatly [11]. The main objective of this study
was to apply and analyze gene expression of thyreotropin
receptor [thyroid stimulating hormone receptor (TSHRmRNA)]
gene and thyroglobulin (Tg) gene in peripheral
blood of healthy individuals and patients with DTC, using
RT-PCR and relative quantification with the 2–ΔΔCt method.
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