Bugalho M J, Domingues R S, Pinto A C, Garrão A, Catarino A L, Ferreira T, Limbert E, Sobrinho L
Serviço de Endocrinologia, Instituto Português de Oncologia, Lisboa, Portugal.
Eur J Endocrinol. 2001 Oct;145(4):409-13. doi: 10.1530/eje.0.1450409.
Recent studies have assigned clinical significance and prognostic value to the detection of thyroglobulin (Tg) mRNA in the blood of patients subjected to total thyroidectomy for a papillary or follicular thyroid carcinoma. In this study, we investigated the diagnostic specificity of Tg mRNA detection, analysing blood samples from healthy volunteers and from patients previously subjected to total thyroidectomy for reasons other than a carcinoma of the follicular epithelium.
Total RNA was extracted from whole blood, reverse-transcribed and the cDNA amplified for Tg and glyceraldehyde-3-phosphate dehydrogenase with specific primers. Expression levels were analysed by using a semi-quantitative PCR. In a few cases, Lymphoprep gradients were used to separate the mononuclear and polymorphonuclear cells prior to further analysis by reverse transcription/PCR.
Our data suggested that all individuals expressed Tg mRNA. Moreover, no differences in the expression levels between subjects with and without thyroid glands were documented. Documentation of Tg expression by the mononuclear and polymorphonuclear layers in patients without thyroid glands support the hypothesis that both lymphocytes and granulocytes express Tg and may justify a background expression in blood, independently of the presence of follicular cells in circulation.
Tg mRNA expression is not limited to follicular cells of the thyroid gland, and its expression by normal blood cells should be considered in tests performed for diagnostic purposes.
最近的研究已赋予甲状腺球蛋白(Tg)mRNA检测以临床意义和预后价值,该检测针对因乳头状或滤泡状甲状腺癌接受全甲状腺切除术的患者血液进行。在本研究中,我们通过分析健康志愿者以及之前因滤泡上皮癌以外的原因接受全甲状腺切除术的患者的血样,来研究Tg mRNA检测的诊断特异性。
从全血中提取总RNA,进行逆转录,并使用特异性引物对Tg和甘油醛-3-磷酸脱氢酶的cDNA进行扩增。通过半定量PCR分析表达水平。在少数情况下,在通过逆转录/PCR进行进一步分析之前,使用淋巴细胞分离液梯度来分离单核细胞和多形核细胞。
我们的数据表明,所有个体均表达Tg mRNA。此外,有甲状腺和无甲状腺的受试者之间的表达水平没有差异。无甲状腺患者单核细胞层和多形核细胞层中Tg表达的记录支持了淋巴细胞和粒细胞均表达Tg的假说,并且可能解释了血液中的背景表达,而与循环中滤泡细胞的存在无关。
Tg mRNA的表达不限于甲状腺滤泡细胞,在进行诊断性检测时应考虑正常血细胞对其的表达。