Stage migration and treatment outcome in carcinoma tongue – A comparison of seventh and eighth AJCC pathological staging systems

Background: The eighth edition of the American Joint Committee on Cancer (AJCC) for oral cancer has incorporated additional pathological features like depth of invasion (DOI) and extranodal extension (ENE) into T and N staging. The incorporation of these two factors will impact the staging and, hence, the treatment decisions. The aim of the study was to clinically validate the new staging system in predicting the outcome in patients treated for carcinoma oral tongue. The study also examined the correlation of pathological risk factors with survival. Methods: We studied 70 patients with squamous cell carcinoma of the oral tongue who underwent primary surgical treatment at a tertiary care center in the year 2012. All these patients were restaged pathologically according to the new AJCC eighth staging system. The 5-year overall survival (OS) and disease-free survival (DFS) were calculated using the Kaplan–Meier method. Akaike information criterion and concordance index were calculated between both staging systems to identify a better predictive model. Log-rank test and univariate Cox regression analysis were conducted to find out the significance of different pathological factors on outcome. Results: Incorporation of DOI and ENE resulted in 47.2% and 12.8% stage migration, respectively. DOI less than 5 mm was associated with a 5-year OS and DFS of 100% and 92.9%, respectively, compared to 88.7% and 85.1%, respectively, when the DOI was more than 5 mm. Presence of lymph node involvement, ENE, and perineural invasion (PNI) were associated with inferior survival. The eighth edition had lower Akaike information criterion and improved concordance index values compared with the seventh edition. Conclusion: The eighth edition of AJCC allows better risk stratification. Restaging of cases based on the eighth edition AJCC staging manual resulted in significant upstaging with difference in survival.

A Retrospective Analysis of Disease-free Survival of 2332 Cases of Cancer of the Oral Tongue from a Single Institute in Eastern India from 1998-2017

Background : Over half the patients of Oral Tongue Cancers in India present with locally advanced disease and Nodal Metastasis. Additionally many of them avoid surgical intervention due to fear or belief that Cancer is a Death Sentence.Materials and Methods : A Retrospective analysis of all Oral Tongue Cancer patients treated at Mahavir Cancer Sansthan from 1998 to 2017 was done. The primary aim was to find the Disease Free Survival (DFS) rates of these patients. The secondary aim was to examine if surgical excision improved DFS rates. Results : The mean DFS for all stages was 51 months varying from 90 months in stage 1 to 30 months in stage 4. One in every three patients survived without recurrence of disease for more than five years. The addition of surgical excision at any stage of Cancer, when possible, resulted in a significant increase in DFS.

Estudo imuno-histoquímico e in vitro de proteínas do sistema ativador de plasminogênio em carcinomas de células escamosas de língua oral / Immunohistochemical and in vitro study of plasminogen activator system proteins in oral tongue squamous cell carcinomas

O carcinoma de células escamosas (CCE) é a neoplasia maligna mais frequente da cavidade oral e apresenta prognóstico desfavorável. Assim sendo, pesquisas têm buscado esclarecer o papel de biomarcadores no comportamento biológico do CCE oral. Nesta perspectiva, destacam-se o ativador de plasminogênio tipo uroquinase (uPA) e seu receptor (uPAR), além do inibidor do ativador de plasminogênio-1 (PAI-1). O presente trabalho analisou, por meio de imuno-histoquímica, a expressão das proteínas uPA, uPAR e PAI-1 no CCE de língua oral (CCELO) e sua relação com parâmetros clinicopatológicos. Este experimento também avaliou os efeitos in vitro da proteína recombinante humana PAI-1 (rhPAI-1) na linhagem celular SCC25, derivada de CCELO. A imunoexpressão de uPA, uPAR e PAI-1 foi analisada em 60 casos de CCELO, de forma semiquantitativa, nas células neoplásicas do front de invasão tumoral. Visando a associação dos achados imuno-histoquímicos com variáveis clinicopatológicas e taxas de sobrevida, os casos foram classificados nas categorias baixa expressão (≤50% das células positivas) e alta expressão (>50% das células positivas). No experimento in vitro, foram analisados os seguintes grupos: G0 (controle; células cultivadas na ausência de rhPAI-1), G10 (células tratadas com rhPAI-1 a 10 nM) e G20 (células tratadas com rhPAI-1 a 20 nM). Diferenças entre estes grupos foram investigadas através dos ensaios: viabilidade celular (Alamar Blue), ciclo celular (marcação com iodeto de propídio, PI), apoptose/necrose (marcação com Anexina V e PI), atividade migratória (Wound healing) e invasão celular (Transwell). A análise imuno-histoquímica revelou alta expressão do uPA na maioria dos CCELOs, mas sem relações significativas com parâmetros clinicopatológicos. As expressões do uPAR e do PAI-1, em nível membranar, foram associadas a recidivas locais (p=0,019) e ao elevado tumor budding (p=0,046), respectivamente. A expressão membranar do PAI-1 também apresentou associação significativa com o alto escore de risco histopatológico (p=0,043). A análise estatística evidenciou ausência de associações significativas entre as variáveis imunohistoquímicas (uPA, uPAR e PAI-1) e indicadores de prognóstico do CCELO (sobrevida específica e sobrevida livre da doença). No estudo in vitro, decorridas 24 horas da administração da rhPAI-1, os grupos G10 e G20 exibiram maior viabilidade celular em comparação ao grupo controle (p=0,020), assim como aumento da progressão para a fase S do ciclo celular (p=0,024). No que concerne aos percentuais de células apoptóticas e necróticas, não foram encontradas diferenças significativas entre os grupos. Nos grupos celulares cultivados na presença da rhPAI1, também foi constatado aumento da atividade migratória (p=0,039) e do potencial de invasão (p=0,039), respectivamente, nos intervalos de 24 horas e 72 horas. Os achados deste estudo sugerem o envolvimento das proteínas uPA, uPAR e PAI-1 na patogênese do CCELO. Entretanto, a expressão destes biomarcadores pode não estar relacionada com a sobrevida dos pacientes. Os resultados in vitro demonstram que o PAI-1 exerce efeitos estimulatórios na proliferação, migração e invasão celular, podendo assim contribuir para a agressividade biológica do CCELO (AU).

Squamous cell carcinoma (SCC) is the most frequent malignant neoplasm of the oral cavity and has an unfavorable prognosis. Thus, studies have sought to clarify the role of biomarkers in the biological behavior of oral SCC. Within this context, urokinase-type plasminogen activator (uPA) and its receptor (uPAR), as well as plasminogen activator inhibitor 1 (PAI-1), are particularly interesting. The present study analyzed, by means of immunohistochemistry, the expressions of uPA, uPAR and PAI-1 in oral tongue SCC (OTSCC) and their relationship with clinicopathological parameters. This experiment also evaluated the in vitro effects of recombinant human PAI-1 (rhPAI-1) on the OTSCC-derived cell line SCC-25. The immunoexpression of uPA, uPAR and PAI-1 was analyzed semiquantitatively in neoplastic cells of the invasion front of 60 OTSCC cases. Aiming to determine the association between immunohistochemical findings, clinicopathological variables and survival rates, the cases were classified as low expression (≤50% of positive cells) and high expression (>50% of positive cells). The following groups were analyzed in the in vitro experiment: G0 (control; cells cultured in the absence of rhPAI-1), G10 (cells treated with 10 nM rhPAI-1), and G20 (cells treated with 20 nM rhPAI-1). Differences between these groups were investigated using the following assays: cell viability (Alamar Blue), cell cycle (staining with propidium iodide, PI), apoptosis/necrosis (staining with Annexin V and PI), migratory activity (Wound healing), and cell invasion (Transwell). Immunohistochemical analysis revealed high expression of uPA in most OTSCC cases, but there were no significant associations with clinicopathological parameters. The high membrane expression of uPAR and PAI-1 was associated with local recurrence (p=0.019) and high tumor budding (p=0.046), respectively. Membrane expression of PAI-1 also presented a significant association with high-risk cases (p=0,043). Statistical analysis demonstrated no significant associations between the immunohistochemical variables (uPA, uPAR and PAI-1) and prognostic indicators of OTSCC (disease-specific and disease-free survival). In the in vitro experiment, 24 hours after administration of rhPAI-1, G10 and G20 exhibited greater cell viability compared to the control group (p=0.02), as well as increased progression to the S phase of the cell cycle (p=0.024). There were no significant differences in the percentages of apoptotic or necrotic cells between groups. In the groups cultured in the presence of rhPAI-1, migratory activity (p=0.039) and invasion potential (p=0.039) were found to be increased after 24 and 72 hours, respectively. The findings of this study suggest the involvement of uPA, uPAR and PAI-1 in the pathogenesis of OTSCC. Nevertheless, the expression of these biomarkers may not be related to survival of patients. The in vitro results suggest that PAI-1 exerts stimulatory effects on cell proliferation, migration and invasion and may therefore contribute to the biological aggressiveness of OTSCC (AU).

Análise da expressão imunoistoquímica de regy e de proteínas envolvidas na regulação da apoptose em carcinomas epidermoides de língua oral / Analysis of the immunohistochemical expression of regy and of proteins involved in the regulation of apoptosis in oral tongue squamous cell carcinomas

O câncer na cavidade oral é uma das lesões malignas mais frequentes na população mundial. Como o processo de desenvolvimento das neoplasias malignas remete a danos que promovem um desequilíbrio na regulação da divisão e morte celular, nos últimos anos, diversos estudos foram realizados com o intuito de verificar a influência desses danos no comportamento global das células e na evolução da doença. Nesse contexto, pesquisas recentes mostraram que alterações na expressão de REG podem desempenhar um importante papel na progressão tumoral de várias neoplasias malignas, por interferir na regulação da apoptose. Diante disso, o objetivo desse trabalho foi investigar a expressão imunoistoquímica dos marcadores REG, p53, MDM-2, Bcl-2 e Bax em carcinomas epidermoides de língua (CELs) oral, com a finalidade de promover uma análise comparativa da imunoexpressão destas proteínas com os parâmetros clínico-patológicos de agressividade da lesão, no intuito de identificar se o REG contribui para a progressão do tumor e se interfere na expressão das proteínas relacionadas a apoptose. Para tanto, foram coletadas informações clínicas de 58 pacientes acometidos por CELs. Em seguida, foi realizada análise histopatológica e imunoistoquímica dos marcadores supracitados, em amostras de material biológico parafinado da lesão. Os resultados mostraram que os tumores com metástase nodal e de alto grau histopatológico de malignidade apresentavam percentuais significativamente menores de REG (p<0,05). Em adição, a análise da expressão de p53, MDM-2 e Bax nos diferentes parâmetros clínico-patológicos avaliados nesse trabalho, não revelou diferenças significativas nos percentuais de imunopositividade desses marcadores. Com relação ao Bcl-2, foi visto que tumores de alto grau de malignidade e com óbitos relacionados a doença apresentavam percentual significativamente menor de positividade dessa proteína (p<0,05). Por fim, o teste de correlação de Spearman demonstrou existir fraca correlação positiva, estatisticamente significativa, entre os percentuais de REG e das proteínas MDM-2 e Bcl-2. Baseado nesses achados, pode-se concluir que a redução da expressão de REG parece contribuir para a progressão do CEL oral e pode influenciar na expressão das proteínas relacionadas a regulação da apoptose (AU).

Cancer of the oral cavity is one of the most common malignancies in the world. Since the development of malignant neoplasms is related to damage that causes an imbalance in the regulation of cell division and cell death, many studies have been conducted in recent years to verify the influence of this damage on overall cell behavior and on the progression of the disease. Within this context, recent studies suggest that changes in the expression of REG play an important role in the progression of different malignant tumors by interfering with the regulation of apoptosis. Therefore, the objective of the present study was to investigate the immunohistochemical expression of REG, p53, MDM-2, Bcl-2 and Bax in oral tongue squamous cell carcinoma (OTSCC), correlating the immunoexpression of these proteins with clinicalpathological parameters of tumor aggressiveness, in order to determine whether REG contributes to tumor progression and interferes with the expression of apoptosisrelated proteins. For this purpose, the clinical data of 58 patients with OTSCC were collected and paraffin-embedded tumor specimens were submitted to histopathological analysis and immunohistochemistry using the markers cited above. The results showed significantly higher expression of REG (p<0.05) in low-grade tumors and without lymph node metastases. In addition, comparison of the expression patterns of p53, MDM-2 and Bax according to the clinical-pathological parameters studied revealed no significant differences in the percentage of immunostaining for these markers. Regarding Bcl-2, a significantly lower percentage of immunostaining for this protein was observed in tumors with a high grade of malignancy and tumors associated with deaths related to the disease (p<0.05). Finally, Spearman's correlation test demonstrated a significant weak positive correlation between the percentages of REG and of MDM-2 and Bcl-2. These findings suggest that the reduced expression of REG contributes to the progression of OTSCC and may influence the expression of proteins related to the regulation of apoptosis (AU).

Construction of epithelial membrane protein 1 eukaryotic expression vector and its influence on migration and invasion of human oral tongue squamous carcinoma cells / 华西口腔医学杂志

<p><b>OBJECTIVE</b>This study aimed to construct a eukaryotic expression vector pEGFP-N1-EMP1 of epithelial mem-brane protein 1 (EMP1) and investigate its influence on migration and invasion of human oral tongue squamous carcinoma cells.</p><p><b>METHODS</b>The human EMP1 gene was amplified by reverse transcription polymerase chain reaction and then ligated into the pEGFP-N1 vector by double restriction endonuclease digestion to construct pEGFP-N1-EMP1 recombinant plasmid. After sequencing identification, pEGFP-N1-EMP1 recombinant plasmid and pEGFP-N1 plasmid were transfected into human oral tongue squamous carcinoma Tb3.1 cell line. The expression of green fluorescent protein in cells was observed after transfection using an inverted fluorescence microscope. The overexpression of EMP1 mRNA was identified at 24, 48, and 72 h after transfection by real-time fluorescence quantitative polymerase chain reaction. The effect of EMP1 overexpression on migration and invasion of Tb3.1 cells was detected by Transwell assay.</p><p><b>RESULTS</b>The full-length EMP1 gene sequence was successfully obtained. Sequence analysis showed that the EMP1 gene was inserted into the pEGFP-N1 vector correctly. Green fluorescence was observed in the transfected cells under fluorescence microscopy. The results of real-time fluorescence quantitative polymerase chain reaction indicated that the expression of EMP1 at 24 h after pEGFP-N1-EMP1 transfection was significantly higher than the other groups. Transwell assays indicated that overexpression of the EMP1 gene could significantly inhibit the migration and invasion ability of Tb3.1 cells.</p><p><b>CONCLUSIONS</b>The eukaryotic expression vector of EMP1 was successfully constructed, and EMP1 overexpression was confirmed to inhibit the migration and inva-sion of oral tongue squamous carcinoma cells in vitro. This study laid a foundation for further investigation on the influence of the EMP1 gene on the metastasis of oral tongue squamous carcinoma and its molecular mechanism.
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A Case of Solitary Angiokeratoma of the Oral Tongue Tip / 대한이비인후과학회지

Angiokeratoma is a rare, cutaneous disorder that typically manifests as multiple lesions and is often associated with a number of metabolic disorders. Although solitary cutaneous forms have been reported, localized lesions within the oral tongue have not been described so far. We report a 16-year-old male with a solitary pigmented lesion of his oral tongue tip. The lesion was a mixture of purple and white in color with a granulomatous appearance. Histopathological examination of the lesion obtained by excisional biopsy showed features consistent with angiokeratoma. We report a case of solitary angiokeratoma of the oral tongue tip and review the clinical and pathologic features of this unusual condition.

Prognostic Value of Volume-Based 18F-Fluorodeoxyglucose PET/CT Parameters in Patients with Clinically Node-Negative Oral Tongue Squamous Cell Carcinoma

OBJECTIVE: To evaluate the prognostic value of volume-based metabolic parameters measured with 18F-fluorodeoxyglucose (18F-FDG) positron emission tomography (PET) in patients with clinically node-negative (cN0) oral tongue squamous cell carcinoma (OTSCC) as compared with other prognostic factors. MATERIALS AND METHODS: In this study, we included a total of 57 patients who had been diagnosed with cN0 tongue cancer by radiologic, 18F-FDG PET/CT, and physical examinations. The maximum standardized uptake value (SUVmax), average SUV (SUVavg), metabolic tumor volume (MTV), and total lesion glycolysis (TLG) for primary tumors were measured with 18F-FDG PET. The prognostic significances of these parameters and other clinical variables were assessed by Cox proportional hazards regression analysis. RESULTS: In the univariate analysis, pathological node (pN) stage, American Joint Committee on Cancer (AJCC) stage, SUVmax, SUVavg, MTV, and TLG were significant predictors for survival. On a multivariate analysis, pN stage (hazard ratio = 10.555, p = 0.049), AJCC stage (hazard ratio = 13.220, p = 0.045), and MTV (hazard ratio = 2.698, p = 0.033) were significant prognostic factors in cN0 OTSCC patients. The patients with MTV > or = 7.78 cm3 showed a worse prognosis than those with MTV < 7.78 cm3 (p = 0.037). CONCLUSION: The MTV of primary tumor as a volumetric parameter of 18F-FDG PET, in addition to pN stage and AJCC stage, is an independent prognostic factor for survival in cN0 OTSCC.

Factors Related to Regional Recurrence in Early Stage Squamous Cell Carcinoma of the Oral Tongue

OBJECTIVES: This study analyzed various clinical and histopathologic factors for patients with early stage squamous cell carcinoma (SCC) of the oral tongue to define a high risk group for regional recurrence and finally to find out the indication of elective neck dissection (END). METHODS: Retrospective chart review was performed for 63 patients with T1-T2N0 SCC of the oral tongue who underwent partial glossectomy with/without END. Clinical and histopathologic factors assessed were age, gender, clinical T stage, tumor cell differentiation, depth of invasion, pathologic nodal status, and intrinsic muscle involvement, perineural invasion, lymphovascular emboli and resection margin involvement. RESULTS: Five year overall survival rate was 97.1% in stage I and 76.2% in stage II, and 5-yr disease free survival rate was 76.7% in stage I and 43.5% in stage II. Rates of occult nodal metastasis in stage I and II were 15.4% and 42.9%, respectively. Overall regional recurrence rate was 15.9%, which consisted of 10.2% in stage I and 35.7% in stage II. The success rate of salvage treatment was 100% in stage I and 40% in stage II. Higher T stage, higher histologic grade, depth of invasion > or =3 mm, presence of intrinsic muscle involvement were significantly related to regional recurrence (P=0.035, P=0.011, P=0.016, P=0.009, respectively). In stage I, the non-END group (n=36) showed 13.9% of regional recurrence rate, while END group (n=13) did not have any regional recurrence (P=0.198). Five year disease free survival rate of END group was significantly higher than non-END group (100% and 68.7%, respectively, P=0.045). CONCLUSION: We recommend to perform END in early stage SCC of the oral tongue if the primary tumor has T2 stage, and T1 stage with higher histologic grade, depth of invasion more than 3 mm, or presence of intrinsic muscle involvement.

Expression of E-Cadherin and Matrix Metalloproteinase-2 in Squamous Cell Carcinoma of the Oral Tongue / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: To evaluate the relationship of the clinicopathologic findings in squamous cell carcinomas of the oral tongue, we studied the expression pattern of E-cadherin, and matrix metalloproteinase-2 (MMP-2) by immunohistochemical stain. MATERIALS AND METHOD: Twenty-five patients with squamous cell carcinomas of the oral tongue were classified as their tumor stage, lymph node metastasis and histopathological grade and their paraffin-embedded surgical specimens were investigated by immunohistochemical analysis using monoclonal E-cadherin antibody and MMP-2 antibody. RESULTS: There was not any significant correlation between the expression of E-cadherin and the tumor stage (p>0.05), but it showed significantly reduced expression of E-cadherin in moderately or poorly differentiated carcinoma (p=0.034). The expression of MMP-2 increased significantly with respect to the metastasis of the cervical lymph node in early carcinoma (p=0.045) but it was not correlated with primary tumor size or histological differentiation (p>0.05). We cannot find any inverse relationship between E-cadherin and MMP-2 expression in oral tongue squamous cell carcinoma. CONCLUSION: The expression of E-cadherin and MMP-2 may be related the disease processes of squamous cell carcinomas of the oral tongue and will provide the possibility of the its biochemical parameters.

Clinical and Therapeutic Aspects of Squamous Cell Carcinoma of Oral Tongue / 대한치료방사선과학회지

Fourty nine patients with squamous cell carcinoma of oral tongue were reviewed retrospectively for the evaluation of clinical manifestation and for the comparison between therapeutic modalites. The gross shape of the tumor was infiltrative in 22, ulcerative in 12, and ulceroinfiltrative type in 10 patients. Direct extension of the tumor was most commonly to the floor of the mouth. The incidence of nodal metastasis generally increased with tumor stage. 55% of the patients showed neck nodal metastasis at the time of diagnosis. Ipsilateral subdigastric node were most commonly involved, followed by submandibular nodes. The 5-year survival rate of patients treated with surgery and radiotherapy was 58.7% in contrast to 21.6% in radiation alone group. Overall 5-year survival rate was 31%. In radiation alone group, half of the patients in stage I, II were locally controlled. But the local control in stage III, IV was much inferior to early lesions. Especially, of 4 patients combined with implantation tecnhnique, 3 were completely controlled. 5-year survival rate of these implanted patients was 50%, 49.4% of patients treated over 7,000 cGy survived 5 years. This was significant in contrast to 6.4% of the group treated below 7,000 cGy. The most common sites of failures were primary sites. In early lesions primary radiotherapy with implantation would be an appropriate treatment in cancer of oral tongue, operation reserved for radiation failure. Operation and adjuvant radiotherapy is recommended in cases of advanced disease.