Background: Differential diagnosis of follicular thyroid carcinoma (FTC) from follicular
thyroid adenoma (FTA) is often difficult since presence or absence of capsular/vascular
invasion can not be determined by preoperative fine needle aspiration cytology,
and may not be judged unanimously on permanent sections even among experienced pathologists.
Determination of molecular-genetic factors such as trefoil factor 3 (TFF3) mRNA in the follicular thyroid tumors may be useful aid to improve the
accuracy of diagnosis, though it is considered to be unstable and relatively low
concentrated genetic substance. Purpose of our study is to investigate expression
level of TFF3 mRNA
of thyroid follicular tumors using formalin-fixed, paraffin-embedded (FFPE) tissue. Methods: Study population included FFPE sections from 19 FTC cases, 20 FTA cases, 11
adenomatous goiter (G) cases and 12 samples of normal thyroid tissue (N) adjacent
to thyroid tumors. RNeasy FFPE kit was used for extraction of total RNA. Purification
and concentration values were determined by spectrophotometer. Extracted RNA was
used for cDNA synthesis in reverse transcription. Synthesized cDNA subsequently
proceeded for relative quantification of TFF3 mRNA by RT-qPCR using TFF3 primers. Glyceroldehyde-3-phosphate
dehydrogenase (GAPDH) and hypoxanthin phosphorobosyltransferase1 (HPRT1) were used
as control genes. The mean and standard deviation of TFF3 mRNA expression level were analyzed by
software Multiplate RQ. Results: Extraction by the FFPE kit yielded high concentration of RNA in all cases.
Purification values were
 M. Eszlinger, K. Krohn, S. Hauptmann, H. Dralle, T. Giordano and R. Paschke, “Perspectives for Improved and More Accurate Classification of Thyroid Epithelial Tumors,” Journal of Clinical Endocrinology & Metabolism, Vol. 93, No. 9, 2008, pp. 3286-3294.
 M. Pulcrano, H. Boukheris, M. Talbot, B. Caillou, C. Dupuy, A. Virion, V. F. De and M. Schlumberger, “Poorly Differentiated Follicular thyroid carcinoma: Prognostic Factors and Relevance of Histological Classification,” Thyroid, Vol. 17, No. 7, 2007, pp. 639-646 doi:10.1089/thy.2007.0029
 B. Hamberger, H. Gharib, L. J. Melton 3rd, J. R. Goellner and A. R. Zinsmeister, “Fine-Needle Aspiration Biopsy of Thyroid Nodules. Impact on Thyroid Practice and Cost of Care,” American Journal of Medicine, Vol. 73, No. 3, 1982, pp. 381-384. doi:10.1016/0002-9343(82)90731-8
 J. M. Cerutti, R. Delcelo, M. J. Amadei, C. Nakabashi, R. M. Maciel, B. Peterson, J. Shoemaker and G. J. Riggins, “Apreoperative Diagnostic Test That Distinguishes Benign from Malignant Thyroid Carcinoma Based on Gene Expression,” Journal of Clinical Investigation, Vol. 113, No. 8, 2004, pp. 1234-1242.
 F. Weber, L. Shen, M. A. Aldred, C. D. Morrison, A. Frilling, M. Saji, F Schuppert, C. E. Broelsch, M. D. Ringel and C. Eng, “Genetic Classification of Benign and Malignant Thyroid Follicular neoplasia Based on a Three-Gene Combination,” Journal of Clinical Endocrinology & Metabolism, Vol. 90, No. 5, 2005, pp. 2512-2521. doi:10.1210/jc.2004-2028
 T. Takano, A. Miyauchi, H. Yoshida, K. Kuma and N. Amino, “Decreased Relative Expression Level of Trefoil Factor 3 mRNA to Galectin-3 mRNA Distinguishes Thyroid Follicular carcinoma from Adenoma,” Cancer Letters, Vol. 219, No. 1, 2005, pp. 91-96. doi:10.1016/j.canlet.2004.07.004
 T. Takano, A. Miyauchi, H. Yoshida, K. Kuma and N. Amino, “High-Throughput Differential Screening of mRNAs by Serial Analysis of Gene Expression: Decreased Expression of Trefoil Factor 3 mRNA in Thyroid Follicular carcinomas,” British Journal of Cancer, Vol. 90, No. 8, 2004, pp. 1600-1605. doi:10.1038/sj.bjc.6601702
 M. Eszlinger, K. Krohn, K. Berger, J. Lauter, S. Kropf, M. Beck, D. Fuhrer and R. Paschke, “Gene Expression Analysis Reveals Evidence for Increased Expression of Cell Cycle-Associated Genes and Gq-Protein-Protein Kinase C Signaling in Cold Thyroid Nodules,” Journal of Clinical Endocrinology & Metabolism, Vol. 90, No. 2, 2005, pp. 1163-1170. doi:10.1210/jc.2004-1242
 D. Fuhrer, M. Eszlinger, S. Karger, K. Krause, C. Engelhardt, D. Hasenclever, H. Dralle and R. Paschke, “Evaluation of Insulin-Like Growth Factor II, Cyclooxygenase-2, Ets-1 and Thyroid-Specific Thyroglobulin mRNA Expression in Benign and Malignant Thyroid Tumours,” European Journal of Endocrinology, Vol. 152, No. 5, 2005, pp. 785-790. doi:10.1530/eje.1.01912
 T. Foukakis, A. Gusnanto, A. Y. Au, A. Hoog, W. O. Lui, C. Larsson, G. Wallin and J. Zedenius, “A PCR-Based Expression Signature of Malignancy in Follicular Thyroid Tumors,” Endocrine-Related Cancer, Vol. 14, No. 2, 2007, pp. 381-391. doi:10.1677/ERC-06-0023
 K. Taniguchi, T. Takano, A. Miyauchi, K. Koizumi, Y. Ito, Y. Takamura, M. Ishitobi, Y. Miyoshi, T. Taguchi, Y. Tamaki, K. Kato and S. Noguchi, “Differentiation of Follicular Thyroid Adenoma from Carcinoma by Means of Gene Expression Profiling with Adapter-Tagged Competitive Polymerase Chain Reaction,” Oncology, Vol. 69, No. 5, 2005, pp. 428-435. doi:10.1159/000089998
 K. Krause, M. Eszlinger, O. Gimm, S. Karger, C. Engelhardt, H. Dralle and D. Fuhrer, “TFF3 Based Candidate Gene Discrimination of Benign and Malignant Thyroid Tumours in a Region with Borderline Iodine Deficiency,” Journal of Clinical Endocrinology & Metabolism, Vol. 93, No. 4, 2008, pp. 1390-1393.
 L. Jin, et al., “HMGA2 Expression Analysis in Cytological and Paraffin-Embedded Tissue Specimens of Thyroid Tumors by Relative Quantitative RT-PCR,” Diagnostic Molecular Pathology, Vol. 20, No. 2, 2011, pp. 71-80. doi:10.1097/PDM.0b013e3181ed784d
 M. T. Tetzlaff, et al., “Differential Expression of Mi-RNAs in Papillary Thyroid Carcinoma Compared to Multinodular Goiter Using Formalin Fixed Paraffin Embedded Tissues,” Endocrine Pathology, Vol. 18, No. 3, 2007, pp. 163-173. doi:10.1007/s12022-007-0023-7
 T. Takano, F. Matsuzuka, H. Sumizaki, K. Kuma and N. Amino, “Rapid Detection of Specific Messenger RNAs in Thyroid Carcinomas by Reverse Transcription-PCR with Degenerate Primers: Specific Expression of Oncofetalfibronectin Messenger RNA in Papillary Carcinoma,” Cancer Research, Vol. 57, No. 17, 1997, pp. 3792-3797.
 T. Takano, A. Miyauchi, T. Yokozawa, F. Matsuzuka, G. Liu, T. Higashiyama, S. Morita, K. Kuma and N. Amino, “Accurate and Objective Preoperative Diagnosis of Thyroid Papillary Carcinomas by Reverse Transcription-PCR Detection of Oncofetalfibronectin Messenger RNA in Fine-Needle Aspiration Biopsies,” Cancer Research, Vol. 58, No. 21, 1998, pp. 4913-4917.
 T. Takano, A. Miyauchi, T. Yokozawa, F. Matsuzuka, I. Maeda, K. Kuma and N. Amino, “Preoperative Diagnosis of Thyroid Papillary and Anaplastic Carcinomas by Real-Time Quantitative Reverse Transcription-Polymerase Chain Reaction of Oncofetalfibronectin Messenger RNA,” Cancer Research, Vol. 59, No. 18, 1999, pp. 4542-4545.