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. 2015 Mar-Apr;81(2):158-66.
doi: 10.1016/j.bjorl.2014.12.003. Epub 2014 Dec 27.

Apoptosis and expression of argyrophilic nucleolus organizer regions in epithelial neoplasms of the larynx

Christiana Vargas Ribeiro  1 , Anilton Cesar Vasconcelos  2 , José de Souza Andrade Filho  3
Affiliations

Apoptosis and expression of argyrophilic nucleolus organizer regions in epithelial neoplasms of the larynx

Christiana Vargas Ribeiro et al. Braz J Otorhinolaryngol. 2015 Mar-Apr.

Abstract
in English, Portuguese

Introduction: Occurrence of apoptosis and expression of proliferative markers are powerful tools to establish a prognosis in the follow-up of cancer.

Objective: To evaluate the growth fraction in papillomas and laryngeal squamous cell carcinomas with three degrees of differentiation through apoptosis and the expression of nucleolus organizer regions.

Methods: Retrospective study from which paraffin material was submitted to microtomy and hematoxylin-eosin and silver staining. Stained slides were used to quantify the apoptotic index and the number of nucleolus organizer regions by morphometry.

Results: Apoptosis was significantly more frequent in well differentiated carcinomas and in papillomas, and a higher growth fraction of expressed nucleolus organizer regions and cells that expressed a greater than average number of nucleolus organizer regions were more frequently noted in undifferentiated carcinomas.

Conclusions: Thus, it was possible to verify that a high apoptotic index was associated with a lower chance of tumor differentiation in carcinomas, while a greater number of total nucleolus organizer regions, cells expressing nucleolus organizer regions above average and a higher growth fraction were associated with greater likelihood of abnormal cell proliferation and increased tumor differentiation.

Introdução: A ocorrência de apoptose e a expressão de marcadores proliferativos são ferramentas poderosas no estabelecimento do prognóstico do câncer.

Objetivo: Avaliar a fração de crescimento de papilomas e carcinomas laríngeos de células escamosas, com três graus de diferenciação, através da apoptose e expressão de regiões organizadoras de nucléolo.

Método: Estudo retrospectivo, cujo material emblocado em parafina foi submetido à microtomia e coloração em hematoxilina-eosina, e pela prata. As lâminas coradas foram utilizadas para quantificar o índice apoptótico e o número de regiões organizadoras de nucléolo (NORs) através da morfometria.

Resultados: A apoptose foi significativamente mais frequente em carcinomas bem diferenciados e em papilomas; enquanto que uma maior fração de crescimento, de NORs expressos e de células que expressaram maior número de NORs, foram mais frequentes nos carcinomas indiferenciados.

Conclusões: Foi possível verificar que o índice apoptótico elevado indica menores chances de diferenciação tumoral nos carcinomas, enquanto que um maior número de NORs totais e células expressando NORs acima da média, e uma maior fração de crescimento, determinam maiores chances de proliferação celular anormal e maior diferenciação tumoral.

Keywords: Apoptose; Apoptosis; Carcinoma de células escamosas; Laringe; Larynx; Nucleolus organizer region; Papilloma; Papiloma; Região organizadora do nucléolo; Squamous cell carcinoma.

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Figures

Figure 1
Figure 1
Grade I squamous cell carcinoma (A) Micrograph of grade I squamous carcinoma cells, with the presence of diffuse keratin pearls (CP) (HE, bar = 100 μm). (B) Micrograph of grade I squamous carcinoma cells with various apoptotic cells with condensation of cytoplasm (arrows) and anoikis (*) (HE, bar = 10 μm).
Figure 2
Figure 2
Grade I squamous cell carcinoma. Micrograph of grade I squamous cells, with invasion of connective tissue by tumor cells in the form of very delimited epithelial nests (EN) (HE, bar = 100 μm).
Figure 3
Figure 3
Grade II squamous carcinoma cells. Micrograph of grade II squamous carcinoma cells with connective tissue infiltration by tumor cells (HE, bar = 100 μm).
Figure 4
Figure 4
Grade II squamous carcinoma cells. Micrograph of grade II squamous carcinoma cells with presence of apoptotic cells with condensation of cytoplasm (arrows) and anoikis (*) (HE, bar = 10 μm).
Figure 5
Figure 5
Grade III squamous carcinoma cells. Micrograph of grade III squamous carcinoma cells with apoptotic cells with nuclear hyperchromatacism (arrows) (HE, bar = 10 μm).
Figure 6
Figure 6
Grade III squamous carcinoma cells. Micrograph of grade III squamous carcinoma cells with presence of a few apoptotic cells with condensation of cytoplasm (arrows) and anoikis (*) (HE, bar = 10 μm).
Figure 7
Figure 7
Papilloma. Micrograph of papilloma with exophytic projections extending below the surface of the mucosa (HE, bar = 100 μm).
Figure 8
Figure 8
Papilloma. Micrograph of papilloma with cell apoptosis with condensation of cytoplasm (arrows) and anoikis (*) (HE, bar = 10 μm).
Figure 9
Figure 9
Grade I squamous carcinoma cells with AgNORs. Micrograph of grade I squamous carcinoma cells with AgNORs in the form of isolated lumps or forming large aggregates, with rounded or irregular shapes filling the entire nucleus or present in the nucleolus as satellites (AgNOR, bar = 10 μm).
Figure 10
Figure 10
Grade II squamous carcinoma cells with AgNORs. Micrograph of grade II squamous cell carcinoma containing AgNORs forming rounded clumps filling the entire nucleolus or as small satellites scattered by the nucleus (AgNOR, bar = 10 μm).
Figure 11
Figure 11
Grade III squamous carcinoma cells with AgNORs. Micrograph of grade III squamous carcinoma cells with AgNORs forming dark clumps together and occupying the entire nucleolus or isolated as small satellites in large nuclei (AgNOR, bar = 10 μm).
Figure 12
Figure 12
Papilloma with AgNORs. Micrograph of papilloma with AgNORs forming small clumps present in the nucleus as satellites or occupying the entire nucleolus (AgNOR, bar = 10 μm).
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