Cytology and mRNA Expression Analysis of Fine Needle Aspirates of Thyroid Nodules in an East German Region with Borderline Iodine Deficiency

Abstract

Fine needle aspiration cytology (FNAC) is widely recommended as an important tool for pre-operative identification of malignancy in patients with nodular thyroid disease. To assess the diagnostic contribution of FNAC and the potential of quantitative mRNA analysis in fine needle aspirates in daily practice, we conducted a prospective study in thyroid clinics (n=2) and endocrine practices (n=3), respectively in an East German region with borderline iodine deficiency. Two-hundred and forty-four consecutive FNACs were obtained over a period of 2 years (2002-2004) from euthyroid patients presenting for first evaluation of a solitary thyroid nodule. The mean nodule size for FNAC was 27 mm (range: 10-79 mm). In 55% of patients FNAC was performed after scintiscan detection of a cold or normal functioning thyroid nodule (CTN), while in the remainder FNAC was performed as a primary investigation. FNAC outcomes were: 57.8% benign, 22.1% indeterminate, 2.5% suspicious for malignancy, 17.6% non-diagnostic. Messenger RNA levels for a house keeping gene (β-actin) and a thyroid specific marker (thyroglobulin, Tg) were studied as basic molecular markers using real-time PCR. Both in the in vivo and ex vivo FNA series, β-actin and Tg mRNA levels were positively correlated with the thyrocyte cell yield/respective FNA smear. However, subgroup analysis showed that FNAC with histologically confirmed follicular thyroid cancer and/or microfollicular adenoma exhibited significantly lower Tg mRNA expression despite high β-actin levels. Sufficient mRNA quantities were obtained in >90% of FNA specimen to allow quantitative mRNA analysis of at least 5 further genes. In conclusion, quantitative mRNA analysis is feasible in FNA on a routine basis and provides a perspective for a molecular distinction of thyroid nodules, once specific marker genes have been defined for benign and malignant thyroid tumours respectively.

Thyroid lesion - β-actin - Tg mRNA - scintiscans

References

• 1 Hampel R, Kulberg T, Klein K, Jerichow JU, Pichmann EG, Clausen V, Schmidt I. Goiter incidence in Germany is greater than previously suspected.  Med Klin. 1995;  90 324-329
• 2 Volzke H, Ludemann J, Robinson DM, Spieker KW, Schwahn C, Kramer A, John U, Meng W. The prevalence of undiagnosed thyroid disorders in a previously iodine-deficient area.  Thyroid. 2003;  13 803-810
• 3 Reiners C, Wegscheider K, Schicha H, Theissen P, Vaupel R, Wrbitzky R, Schumm-Draeger PM. Prevalence of thyroid disorders in the working population of Germany: ultrasonography screening in 96,278 unselected employees.  Thyroid. 2004;  14 926-932
• 4 Elisei R, Bottici V, Luchetti F, Di Coscio G, Romei C, Grasso L, Miccoli P, Iacconi P, Basolo F, Pinchera A, Pacini F. Impact of routine measurement of serum calcitonin on the diagnosis and outcome of medullary thyroid cancer: experience in 10,864 patients with nodular thyroid disorders.  J Clin Endocrinol Metab. 2004;  89 163-168
• 5 Karges W, Dralle H, Raue F, Mann K, Reiners C, Grussendorf M, Hufner M, Niederle B, Brabant G. Calcitonin measurement to detect medullary thyroid carcinoma in nodular goiter: German evidence-based consensus recommendation.  Exp Clin Endocrinol Diabetes. 2004;  112 52-58
• 6 Hegedus L, Bonnema SJ, Bennedbaek FN. Management of simple nodular goiter: current status and future perspectives.  Endocr Rev. 2003;  24 102-132
• 7 Gharib H, Goellner JR. Fine-needle aspiration biopsy of the thyroid: an appraisal.  (Review) Ann Intern Med. 1993;  118 282-289
• 8 Bennedbaek FN, Perrild H, Hegedus L. Diagnosis and treatment of the solitary thyroid nodule. Results of a European survey.  Clin Endocrinol. 1999;  50 357-363
• 9 Baloch ZW, Hendreen S, Gupta PK, LiVolsi VA, Mandel SJ, Weber R, Fraker D. Interinstitutional review of thyroid fine-needle aspirations: impact on clinical management of thyroid nodules.  Diagn Cytopathol. 2001;  25 231-234
• 10 Baloch ZW, Sack MJ, Yu GH, LiVolsi VA, Gupta PK. Fine-needle aspiration of thyroid: an institutional experience.  Thyroid. 1998;  8 565-569
• 11 Hamburger JI. Diagnosis of thyroid nodules by fine needle biopsy: use and abuse.  J Clin Endocrinol Metab. 1994;  79 335-339
• 12 Oertel YC. Some thoughts about fine needle aspirations of thyroid nodules.  Thyroid. 2004;  14 86-87
• 13 Oertel YC. A pathologist trying to help endocrinologists to interpret cytopathology reports from thyroid aspirates.  J Clin Endocrinol Metab. 2002;  87 1459-1461
• 14 Dietlein M, Dressler J, Eschner W, Leisner B, Reiners C, Schicha H. Procedure guideline for thyroid scintigraphy (version 2).  Nuklearmedizin. 2004;  42 120-122
• 15 DeLellis R, Heitz P, Lloyd R, Eng C. WHO classification of tumors .Pathology and genetics of tumors of endocrine organs. IARC Press 2004
• 16 Weber F, Shen L, Aldred MA, Morrison CD, Frilling A, Saji M, Schuppert F, Broelsch CE, Ringel MD, Eng C. Genetic classification of benign and malignant thyroid follicular neoplasia based on a three-gene combination.  J Clin Endocrinol Metab. 2005;  90 2512-2521
• 17 Fagin JA. Mini review: branded from the start-distinct oncogenic initiating events may determine tumor fate in the thyroid.  Mol Endocrinol. 2002;  16 903-911
• 18 Takano T, Miyauchi A, Yoshida H, Kuma K, Amino N. Decreased relative expression level of trefoil factor 3 mRNA to galectin-3 mRNA distinguishes thyroid follicular carcinoma from adenoma.  Cancer Lett. 2005;  219 91-96
• 19 Brauer VF, Brauer WH, Fuhrer D, Paschke R. Iodine nutrition, nodular thyroid disease, and urinary iodine excretion in a German university study population.  Thyroid. 2005;  15 364-370
• 20 Hegedus L. Clinical practice. The thyroid nodule.  N Engl J Med. 2004;  351 1764-1771
• 21 Cibas ES, Ducatman BS. Cytology. Diagnostic principles and clinical correlates. 2nd Edition. Saunders 2003
• 22 Führer D, Eszlinger M, Karger S, Krause K, Engelhardt C, Hasenclever D, Dralle H, Paschke R. Evaluation of insulin-like growth factor II, cyclooxygenase-2, ets-1 and thyroid-specific thyroglobulin mRNA expression in benign and malignant thyroid tumours.  Eur J Endocrinol. 2005;  152 785-790
• 23 Orell SR, Philipps J. The Thyroid Fine needle biobsy and cytological diagnosis of thyroid lesions.  Monographys in Clinical Cytology Vo, 14. Karger
• 24 Hedayati N, McHenry CR. The clinical significance of an isofunctioning thyroid nodule.  Am Surg. 2003;  69 311-316
• 25 Singer PA, Cooper DS, Daniels GH, Ladenson PW, Greenspan FS, Levy EG, Braverman LE, Clark OH, McDougall IR, Ain KV, Dorfman SG. Treatment guidelines for patients with thyroid nodules and well-differentiated thyroid cancer. American Thyroid Association.  Arch Intern Med. 1996;  156 2165-2172
• 26 de Vos tot Nederveen Chappel RJ, Bouvy ND, Bonjer HJ, van Muiswinkel JM, Chadha S. Fine needle aspiration cytology of thyroid nodules: how accurate is it and what are the causes of discrepant cases?.  Cytopathology. 2001;  12 399-405
• 27 Mazzaferri EL, Robbins RJ, Spencer CA, Braverman LE, Pacini F, Wartofsky L, Haugen BR, Sherman SI, Cooper DS, Braunstein GD, Lee S, Davies TF, Arafah BM, Ladenson PW, Pinchera A. A consensus report of the role of serum thyroglobulin as a monitoring method for low-risk patients with papillary thyroid carcinoma.  J Clin Endocrinol Metab. 2003;  88 1433-1441
• 28 Nikiforova MN, Lynch RA, Biddinger PW, Alexander EK, Dorn GW, II, Tallini G, Kroll TG, Nikiforov YE. RAS point mutations and PAX8-PPAR rearrangement in thyroid tumors: evidence for distinct molecular pathways in thyroid follicular carcinoma.  J Clin Endocrinol Metab. 2003;  88 2318-2326
• 29 Gardner HA, Ducatman BS, Wang HH. Predictive value of fine-needle aspiration of the thyroid in the classification of follicular lesions.  Cancer. 1993;  71 2598-2603

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