Evaluation of “AgNOR Count / Nucleus” and “Total AgNOR Area / Nuclear Area’’ Proportions in Thyroide Cells of Cases with Nodular Goiter

Year 2012, Volume: 14 Issue: 3, 5 - 10, 01.12.2012

Nurhan Cücer Murat Oktay Yusuf Aydın Recep Eröz

Abstract

Aim: NORs are loops of DNA, tandemly repeated that transcribed into ribosomal RNA (rRNA)processed into preribosomes in the nucleoli, and which eventually become part of matureribosomes in the cytoplasm. These centers are visible in interphase nuclei when examined aftera silver staining. The aim of this study was the comparison of the AgNOR count/nucleus and‘‘Total AgNOR area/ nuclear area’’ (TNA / NA) proportions to different age and sex in caseswith nodular goiter.Materials and Methods: Fifteen female (age range 38–68) and male (age range 38–68) whosefine needle aspiration biopsy (FNAB) were compatible with non-cystic nodular goiter wereincluded in the study. Those biopsy materials obtained by FNAB were stained for AgNORdetection according to a specific protocol. One hundred nuclei per individual have beenevaluated, and both mean AgNOR count and TNA / NA values of individual cells were detectedfor each cases.Results: Male patients with nodular goiter had not significantly higher mean AgNOR count(2.06 ± 0.46 %) than mean AgNOR count of females (1.97 ± 0.49) (p = 0.62). There is not anydifferences between mean TNA / NA ratio of males (6.00 ± 1.11) and females (5.34 ± 1.13) (p= 0.12). Additionally, there is not any correlation between age and mean AgNOR count (r =0.11 p = 0.56) and TNA / NA ratio (r = 0.30; p = 0.11) in both of females and males.Conclusion: The rate of AgNOR proteins synthesis is not effected by age or sex in thyroid cellsof cases with nodular goiter. Therefore, the increment of these proteins in patients is due todisease

Keywords

NOR, AgNORs, nodular goiter, thyroid fine needle aspiration cytology

Nodüler Guatr Tanısı Almış Hastaların Tiroid Hücrelerinin AgNOR Sayısı / Çekirdek ve Toplam AgNOR Alanı / Çekirdek Alanı Oranının Değerlendirilmesi

Abstract

Amaç: Çekirdekçik organize edici bölgeler (NORs) çekirdekçik içerisinde preribozomlarınişlenerek sitoplazmada olgun ribozom parçalarını oluşturan, ribozomal RNA (rRNA)’laratranskribe edilen ve tandem olarak tekrarlanan DNA ilmekleridir. Bu merkezler gümüş boyamasonrasında incelendiğinde, interfaz çekirdeğinde görülebilirler. Bu çalışmanın amacı, nodülerguatır tanısı almış bireylerin tiroid hücrelerindeki ortalama AgNOR sayısı (herbir kişide toplamAgNOR sayısı / incelenen çekirdek sayısı oranı) ve “Toplam AgNOR alanı / çekirdek alanı”(TNA / ÇA) oranlarının cinsiyet ve yaşa göre değişip değişmediğinin araştırılmasıdır.Gereç Yöntem: Çalışmaya ince iğne aspirasyon biyopsisi (İİAB) nodüler guatr ile uyumlu olan(kistik olmayan) 15 Kadın (yaş aralığı 38–68) ve 15 erkek (yaş aralığı 38–68) birey dahil edildi.İİAB ile elde edilen biyopsi materyalleri, AgNOR’ların tespiti için spesifik bir protokole göreboyandı. Her bir birey için 100 çekirdek değerlendirilerek, ortalama AgNOR sayısı ve ‘‘TNA /ÇA’’oranları tespit edildi.Bulgular: Nodüler guatr’lı erkek hastaların ortalama AgNOR sayısı (2,06 ± 0,46), kadınhastaların ortalama AgNOR sayısından (1,97 ± 0,49) anlamlı derecede yüksek değildi (p = 0,62).Erkek hastaların ortalama ‘‘TNA / ÇA’’ oranı (6,00 ± 1,11) ile Kadın hastaların ortalama ‘‘TNA/ ÇA’’oranı (5,34 ± 1,13) arasında da istatistiksel olarak anlamlı bir fark saptanmadı (p = 0,12).Buna ilaveten Kadın hastalarda ve erkek hastalarda hem ortalama AgNOR sayısı ile yaş (r =0,11 p = 0,56) hem de ‘‘TNA / ÇA’’oranı ile yaş arasında anlamlı bir korelasyon yoktu (r = 0,30;p = 0,11).Sonuç: Nodüler guatr tanılı hastaların tiroid hücrelerindeki AgNOR proteinlerinin sentez hızı,yaş veya cinsiyetten etkilenmemektedir. Bundan dolayı, bu proteinlerin hastalarda artışı hastalığabağlıdır

Keywords

AgNORs, NOR, nodüler guatr, tiroid ince iğne aspirasyon sitolojisi

References

• Mennel HD, Müller I. Morphometric investigation on nuclear and nucleolar arrangement and AgNOR content in the rat hippocampus under normal and ischemic conditions. Exp. Toxicol. Pathol. 46:491–501, 1994.
• Ploton D, Menager M, Jeannesson P, Himber G, Pigeon F, Adnet JJ. Improvement in the staining and in the visualization of the argyrophilic proteins of the nucleolar organizer region at the optical level. Histochem. J. 18:5–14, 1986.
• Derenzini M. The AgNORs. Micron 31:117–120, 2000.
• Garcia-Moreno LM, Conejo NM, Pardo HG, Gomez M, Martin FR., Alonso MJ, Arias JL. Hippocampal AgNOR activity after chronic alcohol consumption and alcohol deprivation in rats. Physiol. Behav. 72:115–121, 2001.
• Raska I, Shaw PJ, Cmarko D. New insights into nucleolar architecture and activity. Int. Rev. Cytol. 255:177–235, 2006.
• Sirri V, Urcuqui-Inchima S, Roussel P, Hernandez-Verdun D. Nucleolus: the fascinating nuclear body. Histochem. Cell Biol. 129:13–31, 2008.
• Hayashida M, Miyaoka T, Tsuchie K, Yasuda H, Wake R, Nishida, A, Inagaki T, Toga T, Nagami H, Oda T, Horiguchi J. neuropathological changes in rats: a nucleolar organizer regions in tissue sections and cellular smears. Anal. Quant. Cytol. Histol. 14:14–23, 2009. related behavioral and
• Canepa M, Gambini C, Sementa AR, Borgiani L, Rovida S. Nucleolar organizer regions and Ki-67 immunostaining in ductal breast cancer: a comparative study. Pathologica 82:125-32, 1990.
• Soomro IN, Whimister WF. Growth fraction in lung tumors determined by Ki-67 immunostaining and comparison with AgNOR scores. J Pathol 162:217–22, 1990.
• Trere D. AgNOR staining and quantification. Micron 31.127– 131, 2000.
• Imamoglu N, Demirtas H, Donmez-Altuntas H, Hamurcu Z, Ilten A. NORs Expression increases on metaphase chromosomes of Down syndrome lymphocytes, in concordance with the mitogen concentration in the culture medium. Cytometry Part B-Clin Cytom. 66B:36–39, 2005b.
• Cucer N, Imamoglu N, Tozak H, Demirtas H, Sarac F, Tatlisen A, Ozturk F. Two-dimensional agnor evaluation as a prognostic variable in urinary bladder carcinoma: A different approach via total agnor area/nucleus area per cell. Micron 38.674–679, 2007.
• Eroz R, Cucer N, Karaca Z, Unluhizarci K, Ozturk F. The Evaluation of Argyrophilic Nucleolar Organizing Region Proteins in Fine-Needle Aspiration Samples of Thyroid, Endocr. Pathol. 22.74–78, 2011.
• Eroz R, Cucer N, Unluhizarci K, Ozturk F. Evaluation of AgNOR spot number in thyroid papillary carcinoma and normal cells nuclei. Journal of Health Sciences. 19(2):102- 107, 2010.
• Eroz R, Okur M, Berik O. Down sendromlu çocuklarda AgNOR sayısı ile gelişim arasında bir ilişki var mı? Sağlık Bilimleri Enstitüsü Dergisi 1(1):8-11, 2011.
• Eroz R, Okur M, Ozkan A, Berik O, Gunes C. Does higher NORs expression affect the developmental stages of down syndrome infants? Genetic Counseling 23(2):249-253, 2012.
• Eroz R., Unluhizarcı K, Cucer N, Baltaci D, Oktay M. Kistik Nodüler Guatrlı Olguların Tiroid Hücrelerindeki AgNOR Sayısı ve AgNOR Yüzey Alanı/Çekirdek Alanı Oranının Yaş ve Cinsiyete göre Karşılaştırılması Konuralp Tıp Dergisi 4(2):31-35, 2012.
• Eroz R, Tasdemir S, Dogan H. Is there any relationship between decreased AgNOR protein synthesis and human hair loosing? Biotechnic & Histochemistry Early Online:1–5, 2012.
• Pich A, Chiusa L, Margaria E. Role of the argyrophilic nucleolar organizer regions in tumor detection and prognosis. Cancer Detect Prev 19:282-91, 1995.
• Pich A, Chiusa L, Margaria E. Prognostic relevance of AgNORs in tumor pathology. Micron 31:133-41, 2000.
• Montironi R, Bracischi A, Scarpelli M, Matera G, Alberti R. Value of quantitative nucleolar features in the preoperative cytological diagnosis of follicular neoplasias of the thyroid. J Clin Pathol 44:509-514, 1991.
• Crissmann JD, Drozdowicz S, Johnson C, Kini SR. Fine needle aspiration diagnosis of hyperplastic and neoplastic follicular nodules of the thyroid: a morphometric study. Anal Quant Cytol Histol 13:41-52, 1991.
• Salmon I, Gasperin P, Remmelink M, Rahier I, Rocmans P, Pasteels JL, Heimann R, Kiss R. Ploidy level and proliferative activity measurements in a series of 407 thyroid tumors or other pathologic conditions. Hum Pathol 24(8):912-920, 1993.
• De Micco C, Ruf J, Chrestian MA, Gros N, Henry JF, Carayon P. Immunohistochemical study of thyroid peroxidase in normal, hyperplastic and neoplastic human thyroid tissues. Cancer 67(12):3036-3041, 1991.
• Kotani T, Asada Y, Aratake Y, Umeki K, Yamamoto I, Tokudome R, Hirai K, Kuma K, Konoe K, Araki Y, et al. Diagnostic usefulness of dipeptidyl aminopeptidase IV monoclonal antibody in paraffin-embedded thyroid follicular tumours. J Pathol 168(1):41-45, 1992.
• Ploton D, Visseaux-Coletto B, Canellas J-C, Bourzat C, Adnet JJ, Lechki C, Bonnet N. Semiautomatic quantification of silver-stained nucleolar organizer regions in tissue sections and cellular smears. Anal Quant Cytol Histol 14(1):14- 23, 1992.
• Rüschoff J. Nucleolar organizer regions in pathomorphologic tumor diagnosis. Veroff Pathol 139: 1-144, 1992.
• Benn PA & Perle M. Chromosome staining and banding techniques. In: Rooney D.E. Czepulkowski BH. (eds). Human Cytogenetics, Constitutional Analysis, actical approach, Vol 1. Oxford Univ. Press 91-118, 1986.
• Lindner LE. Improvements in the silver-staining technique for nucleolar organizer regions (AgNOR). Journal of Histochemistry and Cytochemistry 41:439-445, 1993.
• Demirtas H, Imamoglu N, Donmez H, Cucer N, Yilmaz A, Candemir Z. Condensed chromatin surface and NOR surface enhancement in mitogen-stimulated lymphocytes of Down syndrome patients. Ann Genet-Paris 44:77-82, 2001.
• Howell WM. Selective staining of nucleolus organizer regions (NORs). In: Busch, H., Rothblum, L. (Eds.). The Cell Nucleus, Academic Press, New York, pp. 89–143, 1982.
• Pich A, Chiusa L, Navone R. Prognostic relevance of cell proliferation in head and neck tumours. Annals of Oncology 15:1319-1329, 2004.
• Jones AS, Roland NJ, Caslin AW, Cooke TG, Cooke LD, Foster G. A comparison of cellular proliferation markers in squamous cell Carcinoma of head and neck. J Laryngol Otol 108:859-64, 1994.
• Underwood JCE. AgNOR measurements as indices of proliferation, ploidy and prognosis. J Clin Pathol: Mol Pathol 48 M:239-40, 1995.
• Hall P, Crocker J, Watts A, Stansfeld A. A comparison of region non immunostaining Histopathology 12:373-381, 1988. and s lymphoma. in Hodgkin’
• Jan Mohamed M, Armstrong J, Crocker J, Leyland J, Hulten M. The relationship between number of interphase NOR and NOR bearing chromosomes in non Hodgkin’s lymphoma. J Pathol. 158 (1): 3-7, 1989.
• Kini SR, Miller JM, Hamburger JI, Smith-Purslow MJ. Cytopathology of follicular lesions of the thyroid gland. Diagn Cytopathol 1:123-132, 1985.
• Hamburger JI, Hamburger SW. Fine needle biopsy of thyroid nodules: avoiding the pitfalls. NY State J Med 86: 241-249, 1986.
• Andersen JS, Lyon CE, Fox AH, Leung AKL, Lam YW, Steen H et al. Directed proteomic analysis of the human nucleolus. Curr. Biol. 12 (1):1–11, 2002.
• Denton TE, Liem SL, Cheng KM, Barrett JV. The relationship between aging and ribosomal gene activity in humans as evidenced by silver staining. Mech. Ageing Dev. 15 (1):1–7, 1981.
• Das BC, Rani R, Mitra AB, Luthra UK. The number of silver- staining NORs (rDNA) in lymphocytes of newborns and its relationship to human development. Mech. Ageing Dev. 36 (2):117–123, 1986.
• Buys CH, Osinga J, Anders GJ. Age-dependent variability of ribosomal RNA–gene activity in man as determined from frequencies of silver staining nucleolus organizing regions on metaphase chromosomes of lymphocytes and fibroblasts. Mech. Ageing Dev. 11 (1):55–75, 1979.
• Pedrazzini E, Mamaev N, Slavutsky I. Age related decrease of NOR activity in bone marrow metaphase chromosomes from healthy individuals. Mol. Pathol. 51(1): 39–42, 1998.
• Thomas S, Mukherjee AB. A longitudinal study of human age- related ribosomal RNA gene activity as detected by silver-stained NORs. Mech. Ageing Dev. 92(2–3):101–109, 1996.
• Camargo RS, Shirata NK, di Loreto C, Garcia EA, Castelo A, Longatto Filho A. Significance of AgNOR measurement in thyroid lesions. Analysis and Quantitaive Cytology and Histology 28:188-192, 2006.
• Slowinska-Klencka D, Klencki M, Popowicz B, & Levinski A. Multiparameter analysis of AgNOR in thyroid lesions: comparison with PCNA expression. Histol Histopathol 19:785-792, 2004.