Molecular differential diagnosis of follicular thyroid carcinoma and adenoma based on gene expression profiling by using formalin-fixed paraffin-embedded tissues

BMC Medical Genomics

Barbara Jarzab⁸  Ralf Paschke⁷  Dariusz Lange⁹  Steffen Hauptmann⁵  Michal Jarzab⁶  Monika Kowal⁸  Tomasz Tyszkiewicz⁸  Dagmara Rusinek⁸  Ewa Stobiecka⁹  Michal Swierniak²  Tomasz Stokowy⁴  Thomas Musholt³  Markus Eszlinger⁷  Agnieszka Czarniecka¹  Aleksandra Kukulska⁸  Malgorzata Oczko-Wojciechowska⁸  Bartosz Wojtas⁸  Aleksandra Pfeifer⁴ 

[1] Clinic of Oncological and Reconstructive Surgery, Maria Skłodowska-Curie Memorial Cancer Center and Institute of Oncology, Gliwice Branch, Wybrzeże Armii Krajowej 15, Gliwice 44-101, Poland;Genomic Medicine, Department of General, Transplant, and Liver Surgery, Medical University of Warsaw Warsaw, Poland;Department of General, Visceral, and Transplantation Surgery, University Medical Center of the Johannes Gutenberg University Mainz, Germany;Faculty of Automatic Control, Electronics and Computer Science, Silesian University of Technology, Gliwice Poland;Department of Pathology, Martin Luther University Halle-Wittenberg Halle (Saale), Germany;Radiotherapy and Chemotherapy Clinic, Maria Skłodowska-Curie Memorial Cancer Center and Institute of Oncology, Gliwice Branch, Wybrzeże Armii Krajowej 15, Gliwice 44-101, Poland;Division of Endocrinology and Nephrology, University of Leipzig Leipzig, Germany;Department of Nuclear Medicine and Endocrine Oncology, Maria Skłodowska-Curie Memorial Cancer Center and Institute of Oncology, Gliwice Branch, Wybrzeże Armii Krajowej 15, Gliwice 44-101, Poland;Tumor Pathology Department, Maria Skłodowska-Curie Memorial Cancer Center and Institute of Oncology, Gliwice Branch, Wybrzeże Armii Krajowej 15, Gliwice 44-101, Poland

关键词: Formalin-fixed paraffin-embedded blocks;    Microarray;    Gene expression;    Follicular thyroid cancer;    Follicular thyroid adenoma;   

Others  :  1091916
DOI  :  10.1186/1755-8794-6-38

 received in 2013-04-03, accepted in 2013-09-30,  发布年份 2013

【 摘 要 】

Background

Differential diagnosis between malignant follicular thyroid cancer (FTC) and benign follicular thyroid adenoma (FTA) is a great challenge for even an experienced pathologist and requires special effort. Molecular markers may potentially support a differential diagnosis between FTC and FTA in postoperative specimens. The purpose of this study was to derive molecular support for differential post-operative diagnosis, in the form of a simple multigene mRNA-based classifier that would differentiate between FTC and FTA tissue samples.

Methods

A molecular classifier was created based on a combined analysis of two microarray datasets (using 66 thyroid samples). The performance of the classifier was assessed using an independent dataset comprising 71 formalin-fixed paraffin-embedded (FFPE) samples (31 FTC and 40 FTA), which were analysed by quantitative real-time PCR (qPCR). In addition, three other microarray datasets (62 samples) were used to confirm the utility of the classifier.

Results

Five of 8 genes selected from training datasets (ELMO1, EMCN, ITIH5, KCNAB1, SLCO2A1) were amplified by qPCR in FFPE material from an independent sample set. Three other genes did not amplify in FFPE material, probably due to low abundance. All 5 analysed genes were downregulated in FTC compared to FTA. The sensitivity and specificity of the 5-gene classifier tested on the FFPE dataset were 71% and 72%, respectively.

Conclusions

The proposed approach could support histopathological examination: 5-gene classifier may aid in molecular discrimination between FTC and FTA in FFPE material.

【 授权许可】

   
2013 Pfeifer et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

【 参考文献 】

• [1]Franc B, De-la SP, Lange F, Hoang C, Louvel A, De RA, Vilde F, Hejblum G, Chevret S, Chastang C: Interobserver and intraobserver reproducibility in the histopathology of follicular thyroid carcinoma. Hum Pathol 2003, 34:1092-1100.
• [2]Lang W, Georgii A, Stauch G, Kienzle E: The differentiation of atypical adenomas and encapsulated follicular carcinomas in the thyroid gland. Virchows Arch A Pathol Anat Histol 1980, 385:125-141.
• [3]Cheung L, Messina M, Gill A, Clarkson A, Learoyd D, Delbridge L, Wentworth J, Philips J, Clifton-Bligh R, Robinson BG: Detection of the PAX8-PPAR gamma fusion oncogene in both follicular thyroid carcinomas and adenomas. J Clin Endocrinol Metab 2003, 88:354-357.
• [4]Fagin JA, Mitsiades N: Molecular pathology of thyroid cancer: diagnostic and clinical implications. Best Pract Res Clin Endocrinol Metab 2008, 22:955-969.
• [5]Sahin M, Allard BL, Yates M, Powell JG, Wang XL, Hay ID, Zhao Y, Goellner JR, Sebo TJ, Grebe SK, et al.: PPARgamma staining as a surrogate for PAX8/PPARgamma fusion oncogene expression in follicular neoplasms: clinicopathological correlation and histopathological diagnostic value. J Clin Endocrinol Metab 2005, 90:463-468.
• [6]Esapa CT, Johnson SJ, Kendall-Taylor P, Lennard TW, Harris PE: Prevalence of Ras mutations in thyroid neoplasia. Clin Endocrinol (Oxf) 1999, 50:529-535.
• [7]Shi YF, Zou MJ, Schmidt H, Juhasz F, Stensky V, Robb D, Farid NR: High rates of ras codon 61 mutation in thyroid tumors in an iodide-deficient area. Cancer Res 1991, 51:2690-2693.
• [8]Suarez HG, du Villard JA, Severino M, Caillou B, Schlumberger M, Tubiana M, Parmentier C, Monier R: Presence of mutations in all three ras genes in human thyroid tumors. Oncogene 1990, 5:565-570.
• [9]Vasko V, Ferrand M, Di CJ, Carayon P, Henry JF, De MC: Specific pattern of RAS oncogene mutations in follicular thyroid tumors. J Clin Endocrinol Metab 2003, 88:2745-2752.
• [10]Saggiorato E, De PR, Volante M, Cappia S, Arecco F, Dei Tos AP, Orlandi F, Papotti M: Characterization of thyroid 'follicular neoplasms' in fine-needle aspiration cytological specimens using a panel of immunohistochemical markers: a proposal for clinical application. Endocr Relat Cancer 2005, 12:305-317.
• [11]Abulkheir IL, Mohammad DB: Value of immunohistochemical expression of p27 and galectin-3 in differentiation between follicular adenoma and follicular carcinoma. Appl Immunohistochem Mol Morphol 2012, 20:131-140.
• [12]Bryson PC, Shores CG, Hart C, Thorne L, Patel MR, Richey L, Farag A, Zanation AM: Immunohistochemical distinction of follicular thyroid adenomas and follicular carcinomas. Arch Otolaryngol Head Neck Surg 2008, 134:581-586.
• [13]Cochand-Priollet B, Dahan H, Laloi-Michelin M, Polivka M, Saada M, Herman P, Guillausseau PJ, Hamzi L, Pote N, Sarfati E, et al.: Immunocytochemistry with cytokeratin 19 and anti-human mesothelial cell antibody (HBME1) increases the diagnostic accuracy of thyroid fine-needle aspirations: preliminary report of 150 liquid-based fine-needle aspirations with histological control. Thyroid 2011, 21:1067-1073.
• [14]Tan A, Etit D, Bayol U, Altinel D, Tan S: Comparison of proliferating cell nuclear antigen, thyroid transcription factor-1, Ki-67, p63, p53 and high-molecular weight cytokeratin expressions in papillary thyroid carcinoma, follicular carcinoma, and follicular adenoma. Ann Diagn Pathol 2011, 15:108-116.
• [15]Paunovic I, Isic T, Havelka M, Tatic S, Cvejic D, Savin S: Combined immunohistochemistry for thyroid peroxidase, galectin-3, CK19 and HBME-1 in differential diagnosis of thyroid tumors. APMIS 2012, 120:368-379.
• [16]Nikiforova MN, Tseng GC, Steward D, Diorio D, Nikiforov YE: MicroRNA expression profiling of thyroid tumors: biological significance and diagnostic utility. J Clin Endocrinol Metab 2008, 93:1600-1608.
• [17]Weber F, Teresi RE, Broelsch CE, Frilling A, Eng C: A limited set of human MicroRNA is deregulated in follicular thyroid carcinoma. J Clin Endocrinol Metab 2006, 91:3584-3591.
• [18]Rossing M, Borup R, Henao R, Winther O, Vikesaa J, Niazi O, Godballe C, Krogdahl A, Glud M, Hjort-Sorensen C, et al.: Down-regulation of microRNAs controlling tumourigenic factors in follicular thyroid carcinoma. J Mol Endocrinol 2012, 48:11-23.
• [19]Borup R, Rossing M, Henao R, Yamamoto Y, Krogdahl A, Godballe C, Winther O, Kiss K, Christensen L, Hogdall E, et al.: Molecular signatures of thyroid follicular neoplasia. Endocr Relat Cancer 2010, 17:691-708.
• [20]Chevillard S, Ugolin N, Vielh P, Ory K, Levalois C, Elliott D, Clayman GL, El-Naggar AK: Gene expression profiling of differentiated thyroid neoplasms: diagnostic and clinical implications. Clin Cancer Res 2004, 10:6586-6597.
• [21]Hinsch N, Frank M, Doring C, Vorlander C, Hansmann ML: QPRT: a potential marker for follicular thyroid carcinoma including minimal invasive variant; a gene expression, RNA and immunohistochemical study. BMC Cancer 2009, 9:93. BioMed Central Full Text
• [22]Lubitz CC, Gallagher LA, Finley DJ, Zhu B, Fahey TJ III: Molecular analysis of minimally invasive follicular carcinomas by gene profiling. Surgery 2005, 138:1042-1048.
• [23]Takano T, Miyauchi A, Yoshida H, Kuma K, Amino N: High-throughput differential screening of mRNAs by serial analysis of gene expression: decreased expression of trefoil factor 3 mRNA in thyroid follicular carcinomas. Br J Cancer 2004, 90:1600-1605.
• [24]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.
• [25]Fryknas M, Wickenberg-Bolin U, Goransson H, Gustafsson MG, Foukakis T, Lee JJ, Landegren U, Hoog A, Larsson C, Grimelius L, et al.: Molecular markers for discrimination of benign and malignant follicular thyroid tumors. Tumour Biol 2006, 27:211-220.
• [26]Cerutti JM, Delcelo R, Amadei MJ, Nakabashi C, Maciel RM, Peterson B, Shoemaker J, Riggins GJ: A preoperative diagnostic test that distinguishes benign from malignant thyroid carcinoma based on gene expression. J Clin Invest 2004, 113:1234-1242.
• [27]Chudova D, Wilde JI, Wang ET, Wang H, Rabbee N, Egidio CM, Reynolds J, Tom E, Pagan M, Rigl CT, et al.: Molecular classification of thyroid nodules using high-dimensionality genomic data. J Clin Endocrinol Metab 2010, 95:5296-5304.
• [28]Alexander EK, Kennedy GC, Baloch ZW, Cibas ES, Chudova D, Diggans J, Friedman L, Kloos RT, LiVolsi VA, Mandel SJ, et al.: Preoperative diagnosis of benign thyroid nodules with indeterminate cytology. N Engl J Med 2012, 367:705-715.
• [29]Wu ZJ, Irizarry RA, Gentleman R, Martinez-Murillo F, Spencer F: A model-based background adjustment for oligonucleotide expression arrays. J Am Stat Assoc 2004, 99:909-917.
• [30]Slawski M, Daumer M, Boulesteix AL: CMA: a comprehensive Bioconductor package for supervised classification with high dimensional data. BMC Bioinformatics 2008, 9:439. BioMed Central Full Text
• [31]Vandesompele J, De PK, Pattyn F, Poppe B, Van RN, De PA, Speleman F: Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 2002, 3:RESEARCH0034.
• [32]Foukakis T, Gusnanto A, Au AY, Hoog A, Lui WO, Larsson C, Wallin G, Zedenius J: A PCR-based expression signature of malignancy in follicular thyroid tumors. Endocr Relat Cancer 2007, 14:381-391.
• [33]Saxen E, Franssila K, Bjarnason O, Normann T, Ringertz N: Observer variation in histologic classification of thyroid cancer. Acta Pathol Microbiol Scand A 1978, 86A:483-486.
• [34]Lange D, Sporny S, Sygut J, Kulig A, Jarzab M, Kula D, Jarzab B: [Histopathological diagnosis of thyroid cancer in a multicenter trial]. Endokrynol Pol 2006, 57:336-342.
• [35]Karger S, Krause K, Gutknecht M, Schierle K, Graf D, Steinert F, Dralle H, Fuhrer D: ADM3, TFF3 and LGALS3 are discriminative molecular markers in fine-needle aspiration biopsies of benign and malignant thyroid tumours. Br J Cancer 2012, 106:562-568.
• [36]England SK, Uebele VN, Kodali J, Bennett PB, Tamkun MM: A novel K+ channel beta-subunit (hKv beta 1.3) is produced via alternative mRNA splicing. J Biol Chem 1995, 270:28531-28534.
• [37]Lu R, Kanai N, Bao Y, Schuster VL: Cloning, in vitro expression, and tissue distribution of a human prostaglandin transporter cDNA(hPGT). J Clin Invest 1996, 98:1142-1149.
• [38]Gumienny TL, Brugnera E, Tosello-Trampont AC, Kinchen JM, Haney LB, Nishiwaki K, Walk SF, Nemergut ME, Macara IG, Francis R, et al.: CED-12/ELMO, a novel member of the CrkII/Dock180/Rac pathway, is required for phagocytosis and cell migration. Cell 2001, 107:27-41.
• [39]Hamm A, Veeck J, Bektas N, Wild PJ, Hartmann A, Heindrichs U, Kristiansen G, Werbowetski-Ogilvie T, Del MR, Knuechel R, et al.: Frequent expression loss of Inter-alpha-trypsin inhibitor heavy chain (ITIH) genes in multiple human solid tumors: a systematic expression analysis. BMC Cancer 2008, 8:25. BioMed Central Full Text
• [40]Kinoshita M, Nakamura T, Ihara M, Haraguchi T, Hiraoka Y, Tashiro K, Noda M: Identification of human endomucin-1 and -2 as membrane-bound O-sialoglycoproteins with anti-adhesive activity. FEBS Lett 2001, 499:121-126.
• [41]Barden CB, Shister KW, Zhu B, Guiter G, Greenblatt DY, Zeiger MA, Fahey TJ III: Classification of follicular thyroid tumors by molecular signature: results of gene profiling. Clin Cancer Res 2003, 9:1792-1800.
• [42]Eszlinger M, Krohn K, Hauptmann S, Dralle H, Giordano TJ, Paschke R: Perspectives for improved and more accurate classification of thyroid epithelial tumors. J Clin Endocrinol Metab 2008, 93:3286-3294.
• [43]Yu S, Liu Y, Wang J, Guo Z, Zhang Q, Yu F, Zhang Y, Huang K, Li Y, Song E, et al.: Circulating microRNA profiles as potential biomarkers for diagnosis of papillary thyroid carcinoma. J Clin Endocrinol Metab 2012, 97:2084-2092.