PPGPO- Mestrado em Patologia Oral

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Análise imunohistoquímica do Gata-3 e Il-4 em carcinomas epidermóides de lábio inferior e língua oral
(2019-02-28) Dantas, Afonso Nóbrega; Oliveira, Patricia Teixeira de; ; ; Costa, Edja Maria Melo de Brito; ; Queiroz, Lelia Maria Guedes;
Squamous cell carcinoma (SCC) is the most common malignant neoplasm in the oral cavity, with the lower lip and oral tongue being the most frequent sites. Like all malignancies, SCC has etiology is multifactorial, with strong association with smoking, alcoholism and chronic exposure to solar radiation. At present many studies have turned attention to the role of the inflammatory response in the development and tumor growth, mainly with respect to the cellular immune response. The aim of this study is to analyze the expression of GATA-3 and IL-4 in squamous cell carcinoma of the lower lip (SCCLL) and oral tongue (SCCOT). The selected cases were classified according to the WHO histological grading of malignancy and the immunohistochemical analysis of GATA-3 and IL-4 in 57 cases of SCCLL and SCCOT. The results showed statistical significance of GATA-3 in the stromal (p < 0.05) of these lesions and IL-4 in the parenchyma (p < 0.01), in the stroma (p < 0.01) and in the parenchyma and stroma together (p < 0.01). It also showed a positive correlation between the expression of GATA-3 in the parenchyma and IL-4 in the parenchyma (r = 0.438, p = 0.001), between GATA-3 in the parenchyma and IL-4 in the stroma (r = 0.378; p = 0.004), and between GATA-3 in the parenchyma and IL-4 in the parenchyma and stroma (r = 0.348; p = 0.008). Therefore, in all cases, the presence of the Th2 response was observed, implying the opening of new horizons in this area for new studies that may contribute to the understanding of this response in the growth and development of these lesions.
Estudo da imunoexpressão do reg-gamma e da beta catenina em queilites actínicas e carcinomas de células escamosas de lábio inferior
(2019-06-28) Parente, Patrícia Davin Gomes; Pinto, Leão Pereira; ; ; Nonaka, Cassiano Francisco Weege; ; Freitas, Roseana de Almeida;
Actinic cheilitis (QA) is a potentially malignant lesion that occurs mainly in men with light skin and a history of chronic sun exposure. Currently, it is not possible to predict which cases of QA will progress to Squamous Cell Carcinoma (SCC), so some biomolecular markers have been researched. Β-catenin is a multifunctional protein that is involved in cell-cell adhesion processes. The alteration of the cadherin-catenin complex has been demonstrated in SCC and correlated with tumor invasion, metastasis and worse prognosis of patients. REGγ is a proteasome activator that can promote the degradation of multiple proteins including p53 and MDM2. Studies show that REGγ is overexpressed in numerous cancers, suggesting that REGγ overexpression is involved in cancer progression. The aim of this study was to analyze the immunohistochemical expression of β-catenin and REGγ in cases of QA and lower lip squamous cell carcinoma (CCELI), comparing the immunohistochemical findings with the clinical and pathological data, in order to verify if there is any a correlation with tumor progression and whether they act synergistically in this process. Β-catenin and REGγ immunoexpression was analyzed semi-quantitatively in 30 cases of QA and 30 cases of CCELI according to the scores: 0 (no labeling); 1 (1-25% positive cells); 2 (26-50% positive cells); 3 (51-75% positive cells); 4 (> 75% positive cells). For statistical analysis, Mann-Whitney and Spearman tests were performed (p <0.05). Both QAs and CCELIs expressed the β-catenin protein, showing an increase in cytoplasmic and nuclear expression in cases of moderate and severe dysplasias. In CCELIs, membrane β-catenin immunoexpression was higher in cases of low grade malignancy. Both QAs and CCELIs expressed the REG-γ protein but no statistical significance between its expression and the degree of epithelial dysplasia was found, as well as between the immunoexpression of REG-γ and the clinicopathological parameters analyzed in the CCELIs. The results of the present study suggest that REG-γ overexpression and reduction in membrane expression of β-catenin may be important events in lip carcinogenesis. However, we believe that this protein is involved in the process of oral carcinogenesis. In this process, correlating the immunohistochemical expression of β-catenin with the expression of REG-γ, as we did not obtain statistically significant results, we suggest that β-catenin expression may not be directly influenced by the levels of REGγ expression.
Análise da expressão imuno-histoquímica do t-bet e ifn-gama em carcinomas epidermoides de lábio inferior e língua oral
(2019-06-07) Rodrigues, Rodrigo Rodrigues; Santos, Pedro Paulo de Andrade; ; ; Souza, Lelia Batista de; ; Alves, Pollianna Muniz;
The treatment of oral squamous cell carcinomas (OSCC) has advanced in recent years and the understanding of molecular mechanisms has been fundamental. The study of markers such as T-bet, which is a specific cell transcription factor of the immune system, and IFN-γ, which is a cytokine participating in the innate immune response and specific immunity, are promising in the literature for better understanding of carcinoma behavior. In this research, 57 cases of OSCC were evaluated, 27 of them with lower lip and 30 with oral tongue of the Oral Pathology Service of the Federal University of Rio Grande do Norte, being classified according to the histological grading of malignancy of the World Health Organization (WHO) and subsequently subjected to immunohistochemistry and T-Bet was evaluated quantitatively and IFN-γ evaluated semi-quantitatively in both the parenchyma and stroma of the OSCC. We found that the LLSCC were classified mostly as well differentiated, while the moderately differentiated CEL were the most evidenced. Immunohistochemical analysis revealed strong cytoplasmic and nuclear labeling for T-Bet and IFN-γ in both the parenchyma and stroma of the OSCC. In this research, the immunohistochemical expression for T-Bet was higher in OTSCC compared to LLSCC showing a value of p = 0.001 when evaluating T-bet in the parenchyma separately and in the parenchyma and stroma. Already the percentage of immunoreactivity for IFN-γ was higher in LLSCC in relation to OTSCC with values of p = 0.049 and p = 0.040 when evaluated in stroma separately and parenchyma and stroma, respectively. The LLSCC showed a more efficient induction of Th1 response than the OTSCC, corroborating the distinct behavior of these lesions, with greater aggressiveness evident in the OTSCC.
Expressão imuno-histoquímica de proteínas ING em desordens potencialmente malignas
(2019-02-28) Gonçalo, Rani Iani Costa; Queiroz, Lelia Maria Guedes; ; ; Costa, Antonio de Lisboa Lopes; ; Barbosa, Natalia Guimarães;
Potentially malignant disorders (PMD) are lesions that can affect the oral cavity and present a potential for malignant transformation. Molecular biomarkers have been studied to help predict the risk of malignant transformation of these lesions. Among them, the ING (inhibitor of growth) proteins (1-5) are highlighted. The objective of this study is to evaluate the expression of ING-1 and ING-2 proteins in lesions diagnosed as hyperkeratosis and ED (mild, moderate and severe), and to correlate the expression pattern observed with the degree of epithelial dysplasia. This is a cross-sectional, retrospective, semi-quantitative and comparative study. The sample consisted of 60 specimens of hyperkeratosis and epithelial dysplasia (ED), which were morphologically evaluated and reclassified according to the binary classification system of ED proposed by Kujan et al. (2006). The immunoexpression for ING-1 and ING-2 in the studied lesions was evaluated semi-quantitatively from the assignment of scores ranging from 0 to 4, according to the percentage of epithelial cells positive for those markers. For the statistical analysis, the Mann-Whitney and Spearman tests (p ≤ 0.05) were performed. Of the 60 cases analyzed, 37 (61.7%) presented as low-risk lesions and 23 (38.3%) as highrisk lesions. For both ING-1 and ING-2, 93.3% of the dysplastic lesions studied presented cytoplasmic and nuclear marking on epithelial cells, with predominance of positivity in the basal and suprabasal layers. Regarding the immunohistochemical marking scores, there was a predominance of cases with high expression (score 4) for both markers. However, when comparing these scores (nuclear, cytoplasmic and general) with the histological gradation of ED, for both ING-1 and ING-2, no statistically significant difference was observed. Thus, the results of the present study suggest that immunoexpression of ING-1 and ING-2 is not related to the degree of oral ED. However, the high expression observed in these lesions suggests that these proteins would be involved in the oral carcinogenesis process.
Expressão imuno-histoquímica das proteínas NF-kB e VEGF165 no líquen plano oral
(2019-02-28) Silva, Nelmara Sousa e; Costa, Antonio de Lisboa Lopes; ; ; Medeiros, Ana Miryam Costa de; ; Moura, Jamile Marinho Bezerra de Oliveira;
The oral Lichen Planus (LPO) is a relatively common chronic inflammatory disease whose etiopathogenesis is still not fully understood. Clinically the LPO is classified into six well identified standards: plate, Bullous, reticular, papular and atrophic erosive. The objective of this research was to compare the expression of nuclear factor Kappa (NF-κB) and vascular endothelial growth factor (VEGF165) in the LPO. This research consisted of a retrospective cross-sectional study of quantitative character and comparative immunohistochemical expression of expression of NF-κB and descriptive of VEGF165 in specimens of oral Lichen Planus Reticularis (30 cases) and erosion (10 cases) and 15 cases of inflammatory fibrous hyperplasia (control). Then analysis Immunohistochemistry to investigate differences in the expression of NF-κB (number of cores positive) in relation to the number of cells with nuclei in particular regions of the specimen negative, being used the methodology adapted from Rich et al. (2018). In the statistical analysis we used the tests (non-parametric test of Kruskal-Wallis one-way ANOVA with Bonferroni corrections and, with a significance level of 5% (p < 0.05). It was observed that for every case of LPO present in this study, there were marking the VEGF165, both in epithelial tissue as in inflammatory infiltrate cells and blood vessels. In the case of HFI, this markup had lower intensity when compared to that of LPO. For the NF-κB no statistically significant difference was found in the epithelial lining against inflammatory fibrous hyperplasia (control), already in the connective tissue (superficial and deep inflammatory infiltrate) expression in this protein was lower than in the case of LPO about inflammatory fibrous hyperplasia. It can be concluded that the inflammatory response in the LPO would be in a supporting role in pathogenesis, and it would not be the isolated factor responsible for the development and perpetuation of the LPO, and may be crucial to presence of epigenetic changes in the immune system to trigger the disease.
Estudo imunoistoquímico das proteínas CD34 e α-SMA em cistos radiculares e cistos radiculares residuais
(2019-02-28) Freitas, Camila Tatyanne Santos de; Galvão, Hebel Cavalcanti; ; ; Santos, Pedro Paulo de Andrade; ; Nunez, Manuel Antonio Gordon;
Introduction: Radicular cysts are inflammatory lesions resulting from infectious processes of pulp necrosis. Myofibroblasts (MFs) and angiogenesis are important factors in the development and progression of cystic lesions, so that MFs secrete growth factors and proteases, as well as angiogenesis stimulates matrix deposition and cell migration, affecting the growth of periapicopathies. Objective: This study aims to evaluate the immunohistochemical expression of CD34 and α-SMA in radicular cysts (CRs) and residual radicular cysts (CRRs), in order to contribute a better understanding of expansion and progression of periapical lesions. Materials and Methods: Clinical, morphological and immunohistochemical analyzes were performed on 30 CRs and 30 CRRs. The present study constitutes a descriptive, quantitative and comparative analysis of CD34 and α-SMA positive immunohistochemical expression in the specimens. Expression of α-SMA was evaluated in the fibrous capsule, at 100x magnification 10 fields of higher immunolabeling were selected below epithelial lining, thereafter at 400X magnification, the positive cells were quantified and at the end were calculated the averages of positive cells per field. However, angiogenic index was performed by counting microvessels immunolabelled by anti-CD34 antibody in 10 (200x) fields. Statistical analysis was performed using the Mann-Whitney, Pearson's Chi-square test and Spearman's correlation, with significance level set (p <0.05). Results: The results showed that there were statistically significant differences regarding α-SMA and α-SMAxCD34, however with respect to the CD34 immunostaining, there were no statistical differences between the lesions. Conclusion: Myofibroblasts and angiogenic index among studied lesions presented a significant correlation. Thus, they are believed contribute to the repair of the periapicopathies.
Efeito da fotobiomodulação na proliferação de células-tronco da polpa de dentes decíduos esfoliados cultivadas sobre filmes de ácido poliláctico
(2018-02-23) Macedo, Rômulo Augusto de Paiva; Barboza, Carlos Augusto Galvão; ; ; Moura, Carlos Eduardo Bezerra de; ; Miguel, Márcia Cristina da Costa;
Photobiomodulation (PBM) stimulates the proliferation of different cell types, including stem cells. Stem cells from human exfoliated deciduous teeth (SHEDs) present a greater differentiation potential compared to other mesenchymal stem cells, being the focus of several studies in the field of tissue engineering. In order for cells to proliferate and differentiate in vitro it is necessary to develop a favorable microenvironment, which can be provided by a biomaterial that mimics the natural extracellular matrix, with polylactic acid (PLA) being distinguished for this purpose due to its properties of biocompatibility and biodegradability, as well as low cost. The aim of this study was to evaluate the effect of PBM in different doses on the proliferation and viability of SHEDs cultured on two-dimensional PLA scaffolds. SHEDs were cultured on the films and divided into three groups: (C) non-irradiated control; (L1) irradiated with a dose of 1 J/cm2; and (L4) irradiated with a dose of 4 J/cm2. The irradiations were performed with an InGaAlP diode laser, with wavelength of 660 nm and power of 30 Mw, in a single dose. Cell viability and proliferation were evaluated at 24, 48 and 72 h after irradiation by Alamar Blue assay, while cell morphology and distribution on the surface of the films were evaluated by SEM at 72 hours. The quantitative data were submitted to non-parametric statistical tests. Data from Alamar blue assay showed that groups L1 and L4 exhibited greater cell proliferation compared to group C, with significant differences between L1 and C at 48 and 72 h (p<0.0001) and between L4 and C in 24 (p<0.01), 48 (p<0.001) and 72 h (p<0.0001). The percentage of reduction of Alamar blue was significantly higher in L4 compared to L1 at 24 and 72 h (p<0.05). Analysis of the samples by SEM showed that in the irradiated groups the cells had a more homogeneous distribution along the surface of the films and a higher cell density compared to the group C, with this difference being even more evident in the group L4. We conclude that PBM in the studied parameters, especially with the dose of 4 J/cm2, stimulates the proliferation of SHEDs in contact with PLA films, thus constituting a methodological tool with potential application in tissue engineering techniques.
Análise da proteína de choque térmico 27 (HSP27) em carcinomas de células escamosas de língua oral
(2018-02-20) Torres, Ondina Karla Mousinho Rocha; Miguel, Márcia Cristina da Costa; ; ; Pereira, Karuza Maria Alves; ; Freitas, Roseana de Almeida;
Oral tongue squamous cell carcinoma (OTSCC) presents high rates of morbidity and mortality. New methods of early detection and risk assessment have being studied aiming to predict the prognosis of patients and directing a differentiated treatment. In this context, several molecular biomarkers have been investigated for this purpose among them the heat shock protein 27 (HSP27). This study aimed to analyze whether HSP27 has some influence on the OTSCC, in addition to correlating its immunoexpression with clinicopathological parameters and patient survival. The sample consisted of 55 cases of OTSCC and 20 specimens of oral normal mucosa. For the histomorphological study, it was used the malignancy grading systems proposed by WHO in 2005, by Brandwein-Gensler et al. (2005) and by Almangush et al. (2014). The expression of HSP27 was evaluated through the Immunoreactive Scoring System (IRS). To verify the association of HSP27 immunopositivity with the clinicopathological parameters, it was used the Pearson’s chi-square and Fisher’s exact test. The curves for survival analysis were performed using the Kaplan Meier method. For all evaluations, the significance values were set at p<0.05. A higher IRS for the oral normal mucosa was observed when compared to OTSCC cases (p<0.001), indicating that there is a reduction of expression of this protein in these neoplasms, despite this finding is documented in the literature, the involved mechanisms are unclear, so, specific molecular studies are needed to elucidate the genetic and epigenetic events involved in the reduction of HSP27 expression in OTSCC cases. No significant associations were found between the immunostaining of this protein and the clinicopathological parameters and patient survival suggesting that this protein does not seem to influence the prognosis of OTSCC patients.
Imunoexpressão de CLIC4 e α-SMA em carcinoma de células escamosas oral e carcinoma verrucoso oral
(2018-02-26) Xerez, Mariana Carvalho; Costa, Antonio de Lisboa Lopes; ; ; Galvão, Hebel Cavalcanti; ; Morais, Maria de Lourdes Silva de Arruda;
Among the malignant neoplasias affecting the oral cavity, oral squamous cell carcinoma (OSCC) is the most frequent presenting high rates of morbidity and mortality, and oral verrucous carcinoma (OVC), which exhibits a behavior distinct from a low grade variant of oral squamous cell carcinoma. The development and progression of these malignant neoplasms are related to the imbalance in the regulation of cell division and death associated to the tumor microenvironment. CLIC4 protein is related to the regulation of the cell cycle, being supraregulated in response to apoptosis and also participating in the process of transdifferentiation of fibroblasts in cancer-associated fibroblasts (CAF), which now express αSMA. CAFs are the main cellular components of the tumor microenvironment, having been related to the aggressiveness and prognosis of several tumors. The aim of this study was to evaluate the immunoexpression of CLIC4 and α-SMA proteins in oral carcinomas. The sample consisted of 20 cases of OSCC, 15 cases of OVC and 5 cases of OM. From the medical records, clinical data regarding the age, gender and location of the lesion were collected. A morphological analysis was performed on HE and semiquantitative immunohistochemical expression of the CLIC4 and α-SMA proteins in samples of paraffin-shaped lesion material. Pearson's Chi-square test and Fisher's exact test were used to verify associations. The Spearman test was used to search for correlation. Significance was set at of 5%. Results: Among the OSCC, the majority occurred in male patients (n = 11; 55.0%), with a mean age of 66.95 years and the most affected anatomic location was tongue (n = 9; 45.0%). The OVC was the female sex (n = 9; 60.0%), the mean age was 61.71 years, and the alveolar ridge was the most affected (n = 4; 26.7%) and lip mucosa (n = 4; 26.7%). For the analysis of CLIC4 expression, its cellular location was considered. Comparing the lesions for CLIC4 labeling in the nucleus, cytoplasm and nucleus /cytoplasm of tumor epithelial cells, no significant difference was observed. When comparing CLIC4 expression in the OSCC and OVC stroma, a significant difference was observed (p <0.0001). The analysis of α-SMA immunostaining in mesenchymal cells of OSCC and OVC revealed an increase in the expression of this protein in the tumor stroma of both tumors, and no significant difference was observed between the lesions. When comparing the immunoexpression of αSMA and CLIC4 in the stroma of OSCC and OVC, a positive correlation was observed but not significant in OSCC (r: 0.350 and p: 0.130). In the OVC, a positive and significant correlation was observed between the CLIC4 and α-SMA proteins (r: 0.612 and p: 0.015). In conclusion, the results of the present study suggest that the decrease or absence of CLIC4 protein nuclear marking associated with increased expression of this protein in the tumor stroma seems to contribute to the process of carcinogenesis and progression of CCEO and CVO and may have an influence on expression of α-SMA.
Análise comparativa da imunoexpressão de twist e da podoplanina entre carcinomas de células escamosas de língua oral e de lábio inferior
(2018-02-23) Rolim, Larissa Santos Amaral; Pinto, Leão Pereira; ; ; Nonaka, Cassiano Francisco Weege; ; Souza, Lelia Batista de;
Introduction: Oral Squamous Cell Carcinoma (OSCC) is a major public health problem worldwide. It is believed that the mechanism responsible for tumor progression is based on collective invasion of cell groups or individual cells, called Epithelial-Mesenchymal Transition (EMT), where there is a decrease in the expression of epithelial biomarkers and increased expression of mesenchymal biomarkers as Twist. Podoplanin (PDPN), a transmembrane glycoprotein, is assumed to be involved in the motility of neoplastic cells that are undergoing through EMT to guide the cell complex during an invasion. Objective: To perform a comparative analysis of the immunoexpression of PDPN and Twist among 40 cases of lower lip CCEs and 36 cases of oral tongue, (primary tumor size, regional and distal lymph node metastasis, clinical staging, histological grade of malignancy and histological pattern of invasion). Methods: To evaluate the histopathological grade of malignancy, the system proposed by Brandwein-Gensler et al. (2005) was used. For the two markers, four types of immunohistochemical analysis were performed: analysis of the invasion front, tumor compressive areas, large tumor islands (> 15 neoplastic cells) and nests/dissociated cells (<15 neoplastic cells). The non-parametric Mann-Whitney test was used for the analysis of the relationships between the clinical and histopathological parameters and the PDPN and Twist immunoexpressions. To verify possible correlations between the PDPN and Twist immunoexpressions, the Spearman (r) correlation test was performed. For all statistical tests, a significance level of 5% (p <0.05) was established. Results: No statistically significant differences were found between PDPN and Twist immunoexpression and tumor size, metastasis, and clinical staging on lower lip SCC (p > 0.05). However, statistically significant differences were found between the cytoplasmic immunoexpression of PDPN and the invasion patterns type 4 and 5 (p = 0.032) of Brandwein-Gensler et al. (2005). Also, no statistically significant differences were found between PDPN and Twist immunoexpression and tumor size, metastasis and clinical staging on oral tongue SCC (p > 0.05). Statistically significant differences were observed between the cytoplasmic (p = 0.006), membrane (p = 0.030) and general (p = 0.025) PDPN expression in oral tongue SCC with the invasion patterns type 4 and 5 of Brandwein-Gensler et al. (2005). In addition, the Spearman correlation test demonstrated statistically significant negative correlations between PDPN membrane expression and Twist general expression (r = -0.356, p = 0.024) and Twist cytoplasmic expression (r = -0.336; p = 0.034) in the lower lip SCCs. Conclusions: PDPN expression is inversely related to Twist in lower lip SCC, it was demonstrated that both proteins are associatved with a worse invasion pattern in SCC. It has also been hypothesized that the relationship of PDPN with Twist in OSCC may be more involved with a partial transition from the epithelial to the mesenchymal phenotype than a complete transition coordinated by PDPN.
Relação entre mastócitos e e-caderina em ceratocistos odontogênicos e cistos radiculares
(2018-02-27) Pinheiro, Juliana Campos; Santos, Pedro Paulo de Andrade; ; ; Carvalho, Cyntia Helena Pereira de; ; Souza, Lelia Batista de;
Mast cells are secretory cells that release different chemical mediators, among them tryptase, participating in both physiological and pathological stimuli. E-cadherin is an element of the cadherin family known to play an important role in the regulation of intercellular adhesion in epithelial tissues. The present study aimed to evaluate the relationship between mast cells and E-cadherin expression in odontogenic keratocysts and radicular cysts. The immunohistochemical evaluation of tryptase consisted of the quantification of degranulated and non-degranulated mast cells in 15 fields, 5 in the epithelial tissue, 5 in the superficial connective tissue and 5 fields in the deep connective tissue. The evaluation of E-cadherin was performed semi-quantitatively, using the scores: 1 (0 to 50%) and 2 (> 50%), also observing the cellular immunoreactivity pattern (membrane, cytoplasmic, membrane and cytoplasmic) in addition to the verification of the size of the lesions from the data collection in the clinical records of the patients. The analysis of the cystic lesions studied revealed a higher total mean number of mast cells in the radicular cysts when compared to odontogenic keratocysts (95% CIs). With regard to the amount of mast cells in the epithelial component, a higher median was observed in the radicular cysts in relation to odontogenic keratocysts; in the connective tissue, odontogenic keratocysts were the largest median. When verifying the density of degranulated and non-degranulated mast cells through the Kruskal-Wallis non-parametric test, it was found that the highest concentrations were related to degranulated mast cells in the epithelium of radicular cysts (KW = 30,343; p = 0,000). Odontogenic keratocysts the highest densities of these degranulated cell types were found in connective tissue (KW = 0,092; p = 0,762). When examining the expression of E-cadherin in the epithelial component, a similarity was observed between cystic lesions with greater expression in odontogenic keratocysts (p = 0,798), which presented membrane marking in 63.3% of the cases. Spearman's correlation analysis showed a negative correlation between E-cadherin expression and total mast cells (r = -0,311; p = 0,016), number of degranulated mast cells (r = -0,255; p = 0,049) and the location of these cell types in total and deep connective tissue in both radicular cysts and odontogenic keratocysts. There was also another negative correlation between intraepithelial mast cells (r = -0,265; p = 0,016) and cystic lesion size. In view of these findings, we observed that the higher concentration of mast cells in the radicular cysts confirms their inflammatory nature, highlighting the greater amount of degranulated mast cells in the epithelial component, reflecting less e-cadherin expression when compared to odontogenic keratocysts. Possible tryptase activity in the alteration of adhesion joints and consequent increase in epithelial permeability. Regarding the size of the lesions, it was verified that the higher mast cell densities in the epithelial component were identified in smaller lesions, suggesting an action of these cell types in an initial stage of growth of cystic lesions studied.
Imunossupressão das proteínas XPF e XRCC1 em carcinoma epidermoide de língua oral e lábio inferior
(2018-02-23) Moreira, Deborah Gondim Lambert; Freitas, Roseana de Almeida; ; ; Andrade, Ana Luiza Dias Leite de; ; Souza, Lelia Batista de;
The DNA repair system involves genes and proteins essential for the maintenance of genome integrity and consequent control of various cellular processes. Alterations in these genes and proteins are involved in the development of tumors, in their evolution and resistance to radio and chemotherapy treatments. The aim of this study was to analyze the immunoexpression of DNA repair proteins, XPF and XRCC1 in squamous cell carcinoma of the lower lip (SCCLL) and oral tongue (SCCOT) and investigate possible associations with the clinical and histopathological groups. For this purpose, 80 cases of SCCLL and SCCOT were selected and evaluated by grading systems proposed by Brandwein-Gensler et al. (2005) and Almangush et al. (2014). The immunohistochemical expression was performed semi-quantitatively, was used for statistical analysis the Chi-square tests, or when applicable, Fisher's exact test, to evaluate an association between the expressions of the proteins as clinical-pathological characteristics. A high nuclear expression of XPF was observed in 72.5% of cases of SCCOT and in 70% of cases of SCCLL. Regarding the XRCC1 immunoexpression in cases of SCCOT, high expression was observed in 60% of the cases and in the SCCLL 70% presented high nuclear expression, and no cytoplasmic expression was observed. Statistically significant associations were observed between immunoexpression of proteins with some clinical and histopathological parameters (p<0.05), however, there were no associations with the prognosis of the patients. The high expression of XPF and XRCC1 proteins in the cases of SCCOT and SCCLL suggest its importance in development and progression of these tumors. The results of this research indicate that immunoexpression of the studied proteins is not associated with a clinical and histopathological characteristics determinant for the biological tumor behavior.
Análise comparativa da imunoexpressão de OCT4 e CD44 em carcinoma epidermoide de lábio inferior e queilite actínica
(2018-02-22) Sena, Dáurea Adília Cóbe; Souza, Lelia Batista de; ; ; Moura, Jamile Marinho Bezerra de Oliveira; ; Pinto, Leão Pereira;
The carcinogenesis process is characterized by the transformation of a normal cell into a malignant cell, due to cumulative genetic alterations, influenced by the genetic predisposition of the patient, as well as by environmental factors. In the lower lip, exposure to UV radiation is the main etiological factor associated with the development of epidermoid carcinomas and actinic cheilitis. By the hierarchical model theory in carcinogenesis, the development, growth and metastasis of cancer are driven by a small population of tumor stem cells, with multipower ability and self-renewal. The aim of this study was to compare the expression of OCT4 and CD44 in 40 cases of lower lip epidermoid carcinoma (LLEC) and 40 cases of actinic cheilitis (AC), being analyzed the clinicopathological features and immunoexpression of these proteins. This markers can be used to provide information about diagnosis, progression and prognosis in these lesions. OCT4 and CD44 expression was assessed semiquantitatively by percentage of positive epithelial cells (PP) and intensity of expression (IE), resulting in total immunolabeling score (PIT) ranging from 0 to 7. OCT4 immunoexpression was shown to be both nuclear and cytoplasmic for most cases of AC and LLEC, while for CD44 it was membranous. No statistically significant differences were found between OCT4 and CD44 immunoexpression and clinicopathological parameters, except for lymph node metastasis in which there was a decrease in OCT4 expression in the core tumor of cases with lymph node metastasis. There was a negative and statistically significant correlation between the immunoexpression of these markers in the core tumor. Also, decreased CD44 expression was observed in LLEC cases when compared to AC cases. The results of this study suggest a higher participation of these proteins in early stages of carcinogenesis. In addition, the imbalance of OCT4 and CD44 immunoexpression in the tumor core suggests the presence of different subpopulations of neoplastic cells with distinct pluripotency phenotype, associated with different degrees of cell differentiation.
Queilite actínica: índice de análise clínica
(2018-02-23) Medeiros, Cristianne Kalinne Santos; Oliveira, Patricia Teixeira de; ; ; Medeiros, Ana Miryam Costa de; ; Nunez, Manuel Antonio Gordon;
Introduction: Actinic cheilitis (AC) is an inflammatory condition of the lips caused by chronic exposure to ultraviolet radiation and it is considered a potentially malignant disorder by the World Health Organization (WHO). Objective: develop and test a clinical index for Actinic Cheilitis (ACCI). Methodology: Based on a literature review, a questionnaire composed by 35 clinical characteristics of the AC was elaborated, which were selected and associated with a clinical severity score by a group of 52 surgeon-dentists. After establishing exclusion criteria, 16 characteristics were selected to compose the index. Twenty patients were selected to compose our sample after application the ACCI and incisional biopsy. The biopsied specimens were classified according to the histopathological grading for oral epithelial dysplasia of the WHO (EL-NAGGAR et al., 2017) and the Binary System (KUJAN et al., 2006). The association between the clinical characteristics and the grade of histopathological severity of the lesion was analyzed using Fisher's Exact Test (p <0.05). The precision of the index was tested based on the percentage of sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), accuracy and ROC curve to cut-off points 5, 7, 9, and 11. Results: There was a prevalence of males (80 (90%), whites (90%), with occupational sun exposure (75%). The most frequent clinical characteristics were loss of lip / skin demarcation (100%), dryness (85%), white spot (80%) and flaking (65%). There wasn`t statistically significant association between the clinical characteristics of AC and histopathological gradations analyzed. The cut-off point that presented the best performance for the ROC curve and the highest percentage for specificity, PPV and accuracy was the value 11. Conclusion: The cut-off point 11 obtained the best precision for the ACCI, and this index value was indicated for biopsy, since it presented smaller number of false-positive individuals according to the two histopathological grading analyzed.
Estudo da expressão imuno-histoquímica das proteínas ING3 e ING4 em lesões odontogênicas epiteliais benignas
(2018-02-15) Vargas, Yailit Del Carmen Martinez; Queiroz, Lelia Maria Guedes; ; ; Medeiros, Ana Miryam Costa de; ; Oliveira, Denise Helen Imaculada Pereira de;
Odontogenic cysts and tumors represent a heterogeneous group of lesions originated from epithelial or ectomesenchymal tissues from the embryonic development of the dental organ, affecting only the gnathic bones. Considering that these epithelial lesions have a distinct biological behavior, they have become the subject of numerous investigations. INGs genes play an important role in cell cycle control, senescence, DNA replication and repair, cell proliferation and apoptosis in various neoplasms. The aim of this study was to comparatively evaluate the immunohistochemical expression of ING3 and ING4 proteins in a series of cases of benign epithelial odontogenic lesions. The sample consisted of 20 odontogenic keratocysts (OK), 20 ameloblastomas (AM) and 15 adenomatoid odontogenic tumors (AOT). The immunohistochemical staining was evaluated semiquantitatively on each case studied according to the percentage of positively immunostained epithelial cells, which ranged from 0 to 3. For statistical analysis were used the Kruskal-Wallis and Spearman tests (p <0.05). Results: Analysis of ING3 expression was positive in the cytoplasm and nucleus of epithelial cells without statistically significant difference between group of lesions studied. Analysis of ING4 expression revealed positivity in the cytoplasm of epithelial cells of all lesions studied. The cytoplasmic immunostaining score for ING4 revealed a statistically significant difference between the groups of lesions studied, with a higher frequency in AOT (p <0.05). After pairing, there was a significant difference between the AM and OK, AM and AOT (p <0.05). And the analysis of ING4 nuclear expression in the studied lesions revealed loss of nuclear expression, followed by reduced nuclear expression for cases of AM and OK, and positivity in the nucleus of AOT epithelial cells. The nuclear immunostaining score for ING4 revealed a statistically significant difference between the groups of lesions studied, with a higher frequency in AOT (p <0.05). After pairing, we observed a significant difference between AM and OK and between AM and AOT and between OK and AOT (p <0.05). In conclusion, the results of the present study suggest that the regulation of ING3 and ING4 in cell growth indicate that these proteins may play an important role in the development and progression of odontogenic lesions with different biological behaviors.
Imunoexpressão de twist e e-caderina em queilites actínicas
(2018-01-29) Cavalcante, Israel Leal; Medeiros, Ana Miryam Costa de; ; ; Cavalcante, Roberta Barroso; ; Freitas, Roseana de Almeida;
Background: Actinic cheilitis (AC) is a potentially malignant oral lesion (PMOL) that results from chronic exposure to the sun's rays. Currently, it is not possible to predict which cases of AC will progress to squamous cell carcinoma of the lower lip, therefore, some biomolecular markers have been subject of investigation. In this context, Twist protein, a highly conserved transcription factor that belongs to the family of helix-loop-helix acts on the negative regulation of E-cadherin acting as a great regulator of the mesenchymal phenotype. Little is known about the role of Twist in PMOLs, however, recent studies have shown that this transcription factor may be involved in the process of oral carcinogenesis process since its early stages. Aim: This study aims to evaluate the immunoexpression of Twist and E-cadherin in ACs as well as to verify if this expression is associated with the morphological progression of these lesions. Methods: The epithelial immunoexpression of E-cadherin and Twist was analyzed semi-quantitatively in 86 cases of ACs with different degrees of epithelial dysplasia, according to the scores: (1) <25% positive cells; (2) 25 to 50% positive cells; (3) 50 to 75% positive cells and (4) >75% positive cells. Median comparisons of the E-cadherin and Twist expression scores with the actinic cheilitis’ histological grade were performed using the non-parametric Mann-Whitney test. Results: Analysis of Twist and E-cadherin expression in the actinic cheilitis’ epithelium revealed membrane and cytoplasmic immunoreactivity for E-cadherin in all epithelial layers except in the keratin layer. For Twist, nuclear and cytoplasmic immunoreactivity were observed in all epithelial layers, except for the keratin layer. Statistically significant differences were not found in the medians of total E-cadherin and Twist scores and in the different cell compartments analyzed (membrane and cytoplasmic for E-cadherin and nuclear and cytoplasmic for Twist) in relation to the actinic cheilitis’ histological grade (p > 0,05). Conclusion: The results of this study suggest a high immunoexpression of Twist and E-cadherin in cases of ACs, however, these proteins appear not to be associated with morphological progression in the analyzed cases.
Estudo imunoistoquímico das subpopulações de macrófagos em cistos radiculares e granulomas periapicais de acordo com aspectos clínicos e morfológicos
(2018-02-23) França, Glória Maria de; Galvão, Hebel Cavalcanti; ; ; Andrade, Ana Luiza Dias Leite de; ; Santos, Pedro Paulo de Andrade;
Introduction: Radicular cysts (RCs) and periapical granulomas (PGs) develop in response to inflammatory mediators from pulp necrosis. Macrophages play an important role in the etiopathogenesis of these lesions and are involved in the protective response of the host as well as in the perpetuation of inflammatory reactions. The infiltration of macrophages subpopulations M1 and M2 into sites of chronic inflammation allows the phagocytosis of foreign agents, the release of chemical mediators and may also act as antigen-presenting cells. Aim: To identify the subpopulations of macrophages M1/M2 in RCs and PGs, additionally relates them to clinical and morphological aspects. Material and Methods: A clinic, histopathologic and immunohistochemical analysis were performed on 30 RCs and 30 PGs. The M1 macrophages subpopulation was evaluated by the percentages of CD68 immunostaining associated with the TNF-α inflammatory cytokine and the M2 macrophages subpopulation by its specific CD163 antibody. Clinical, radiographic, symptomatology, treatment and morphological parameters of the lesions were collected and a statistical significance level of 95% was adopted for statistical analysis. Results: The ratio of CD68+/CD163+ cells was higher in the RCs (median=1.22, p=0.002), and the highest TNF-α immunostaining scores were found in the RCs (p=0.018); in the PGs, the CD68+/CD163+ ratio was lower and associated with a greater CD163+ immunostaing (median=1.02, p<0.001). In addition, CD68+ cells had higher percentages of immunostaining in RCs with smaller sizes (p=0.034). Conclusion: The increased CD68 immunostaining associated with TNF-α cytokine in RCs suggests the greatest differentiation for a M1 phenotype. The greater CD163 immunostaining in PGs provide greater differentiation for the M2 phenotype. Therefore, the inflammatory state promoted by the M1 macrophages is related to appearance and maintenance of RCs, on the other hand, the immunomodulatory state of the M2 macrophages is related to the development of PGs.
Imunoexpressão das proteínas twist e e-caderina em carcinoma epidermoide de lábio inferior
(2018-02-16) Morais, Everton Freitas de; Freitas, Roseana de Almeida; ; ; Costa, Antonio de Lisboa Lopes; ; Santos, Edilmar de Moura;
Lower lip squamous cell carcinoma (LLSCC) accounts for approximately 30% of all cases of oral cancer, and results mainly from chronic exposure to ultraviolet radiation. E-cadherin and Twist proteins represent biomarkers involved in the epithelial-mesenchymal transition process, a determinant event in the potential for tumor invasion. The objective of this study was to analyze the immunoexpression pattern of Twist and E-cadherin proteins in LLSCC, investigating possible associations with clinical-pathological parameters indicative of tumor behavior, as well as to evaluate the correlation between proteins. We also evaluated the correlation between Twist and E-cadherin proteins in LLSCC cases. For the analysis of histopathological grading systems of malignancy, the systems proposed by Bryne et al. (1992), Brandwein-Gensler et al. (2005) and Almangush et al. (2014). Increased expression of E-cadherin (general, membrane and cytoplasmic) in LLSCC classified in early clinical stages (stage I and II) (p <0.05) and higher expression of E-cadherin (general and membrane) in cases with remission after 5 years of follow-up (p<0.05). No significant relationships of E-cadherin expression were observed with tumor size, regional metastasis, relapse and the histopathological grade of malignancy. In relation to the Twist protein, it was found a greater general, nuclear and cytoplasmic expression of the same in lesions classified in late stages (III and IV), with recurrence and a high degree of malignancy (p<0.05). In addition, a greater overall expression of Twist protein was also found in high-risk lesions according to the system proposed by Brandwein-Gensler et al. (2005). There was also a significant positive correlation between Twist nuclear immunoexpression and cytoplasmic E-cadherin expression (r = 0.261, p = 0.046). In turn, a significant negative correlation was observed between Twist cytoplasmic expression and E-cadherin membrane expression (r = -0.286; p = 0.028). The results of the present study suggest the potential involvement of Twist and E-cadherin proteins in the modulation of events related to the worst prognosis of LLSCC.
Estudos dos aspectos clínico-patológicos e da imunoexpressão das proteínas APE1 e XRCC1 em uma série de leucoplasias orais
(2018-02-20) Barros, Caio César da Silva; Silveira, Ericka Janine Dantas da; Miguel, Marcia Cristina da Costa; ; ; ; Pereira, Karuza Maria Alves; ; Queiroz, Lelia Maria Guedes;
Background: Oral leukoplakia (OL) is the most frequent oral potentially malignant disorder in dental practice. At histopathological examination, most of the OLs exhibit histological changes that may or may not be associated with epithelial dysplasia (ED), however, there is no consensus if the presence of ED is indicative of malignant transformation. DNA repair is an important genetic mechanism related to carcinogenesis. In this process, specific genes and proteins act in the detection and removal of the damage with subsequent DNA repair, highlighting in this research the base excision repair (BER) pathway which APE1 and XRCC1 proteins act. Thus, the aim of this study was to verify if the APE1 and XRCC1 proteins are associated with clinicopathological features. Methods: Forty patients diagnosed with OL were selected, clinical data were collected in the clinical records and during each patient’s follow-up. For each case, morphological analysis was performed, in case where there was ED the epithelial dysplasia grading according with the World Health Organization was performed. The immunohistochemical expression of APE1 and XRCC1 was evaluated in 24 cases of OL and in six cases of normal oral mucosa (NOM). A semiquantitative analysis was performed, where the cases were classified as negative immunoexpression (0-10% of positive cells), low (11-50% of positive cells) and high immunoexpression (> 50% of positive cells). Pearson’s chi-square and Fisher's exact tests were performed to determine the association between protein expression and clinicopathological features (significance of p≤0.05). Results: Of the 40 cases of OL 65% were female with a mean age of 58 years. Thirty patients had tobacco consumption habit. Tongue was the most affected location (27.5%). Thirty-two cases presented homogeneous lesions while 60% and 41.2% of the cases showed multiple lesions and lesions recurrence, respectively. A significant association was observed in the cases graded as moderate/severe ED with the non-homogeneous clinical aspect (p=0.039). APE1 and XRCC1 proteins showed high expression in NOM and OL independent of the presence or absence of epithelial dysplasia. APE1 demonstrated nuclear immunostaining in 60% of cases graded as moderate or severe epithelial dysplasia (p=0.005). Conclusions: The non-homogeneous clinical aspect of OL was a strong indicator of the presence of moderate/severe epithelial dysplasia. The APE1 and XRCC1 proteins do not appear to be related to molecular processes involved in the development and with the clinicopathological features of oral leukoplakias.
Estudo da expressão imuno-histoquímica das proteínas P53, BAX e BCL-2 em tumores benignos derivados do epitélio odontogênico
(2016-02-19) Tenorio, Jefferson da Rocha; Queiroz, Lelia Maria Guedes; ; ; Sobral, Ana Paula Veras; ; Souza, Lelia Batista de;
Os tumores odontogênicos (TOs) representam um grupo muito complexo e heterogêneo de lesões que são oriundos de tecidos que participam da formação do órgão dental. Mesmo quando agrupados em lesões da mesma origem, como aqueles derivados do epitélio odontogênico, os TOs podem apresentar distintos comportamentos biológicos. Tal característica pode estar relacionada com a expressão de proteínas ligadas à apoptose e controle do ciclo celular. Material e método: A amostra foi constituída de 60 TOs sendo 20 Ameloblastomas (AMBs), 20 Tumores odontogênicos ceratocísticos (TOCs) e 20 Tumores odontogênicos adenomatoides (TOAs). Foi realizada a técnica imuno-histoquímica, pelo método da estreptoavidina-biotina-peroxidase, utilizando os anticorpos anti: p53, BCL-2 e BAX. Para análise estatística, foram utilizados os testes Kruskal-Wallis e Spearman (p<0,05). Resultados: Todos os TOs apresentaram marcação para as proteínas estudadas. Não houve associação estatisticamente significativa entre a expressão das proteínas entre os TOs analisados, no entanto foi identificado correlação positiva entre a expressão de p53 e BCL-2 (r=0,200) juntamente com correlação negativa entre p53 e BAX (r=-0,100). Além disso, as proteínas p53 e BAX estiveram expressas de modo similar entre os AMBs e TOCs. Por semelhante modo, a proteína BCL-2 apresentou-se expressa de maneira equivalente entre AMBs e TOAs. Conclusão: Proteínas reguladoras da apoptose bem como proteínas ligadas ao ciclo celular estão envolvidas no desenvolvimento dos tumores odontogênicos e sua expressão diferencial está relacionada com o comportamento biológico dessas lesões.

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