The relevance of miRNAs as promising biomarkers in lip cancer.

OBJECTIVES: This study aimed to analyze the expression of miR-181b, miR-21, miR-31, and miR-345 in actinic cheilitis with and without epithelial dysplasia and lower lip squamous cell carcinomas, and to verify if the deregulated expression of these miRNAs would be indicative of malignant transformation. MATERIALS AND METHODS: The sample was selected from formalin-fixed paraffin-embedded tissues of 19 actinic cheilitis without epithelial dysplasia, 32 actinic cheilitis with epithelial dysplasia, 42 lower lip squamous cell carcinomas, and 10 nonaltered oral mucosa of the lip. The microRNA (miR, miRNA) expression was quantified by real-time RT-PCR and the expression of the selected miRNAs among the groups of actinic cheilitis and lower lip cancer was compared by chi-square. RESULTS: A higher expression of miR-181b, miR-31, and miR-345 was found in actinic cheilitis without epithelial dysplasia in comparison to that in actinic cheilitis with epithelial dysplasia and with lower lip cancer. There were no differences in miR-21 expression between actinic cheilitis and lower lip cancer. Hierarchical clustering analysis showed a tendency for a downregulation of miR-181b, miR-21, miR-31, and miR-345 in most patients with lower lip cancers. CONCLUSIONS: The upregulation of miR-181b, miR-31, and miR-345 expression in actinic cheilitis without epithelial dysplasia and the decrease in the expression of these miRNAs in actinic cheilitis with epithelial dysplasia and in lower lip cancer are potential biomarkers of malignant progression. CLINICAL RELEVANCE: This miRNA signature can help to identify actinic cheilitis with potential to progress to lip cancer.

Association of lymph vessel density with occult lymph node metastasis and prognosis in oral squamous cell carcinoma.

BACKGROUND: The aims of this study were to determine intra (ILVD) and peritumoral (PLVD) lymphatic vessel density (LVD), and to investigate the relationship of LVD with occult metastasis and prognosis. METHODS: Eighty-seven oral squamous cell carcinomas, in clinical stages I or II, arising in the tongue or floor of the mouth were stained with podoplanin. Lymphatic vessels were quantified in intra and peritumoral areas by sequential analysis and hot spot evaluation. Associations of the ILVD and PLVD with clinicopathologic parameters were determined by Chi-square or Fisher's exact test. The 5 and 10-year survival rates were calculated by the Kaplan-Meier and compared using the log-rank test. RESULTS: No significant association was observed between ILVD or PLDV and clinicopathologic variables including occult lymph node metastasis, or clinical follow-up. However, ILVD showed a significant association with regional recurrence (p = 0.040). The perineural invasion was associated with PLVD (p = 0.041). Disease-specific (p = 0.044) and disease-free survivals (p = 0.016) had significant association with PLVD. CONCLUSIONS: The intra or peritumoral lymphatic vessel density had no predictive value for occult lymph node metastasis in the early stages of oral cancer arising in the tongue or floor of mouth.

Moesin expression by tumor cells is an unfavorable prognostic biomarker for oral cancer.

BACKGROUND: Moesin is a member of the ERM (ezrin, radixin and moesin) proteins that participate in cell migration and tumor invasion through transductional signals sent to actin filaments by glycoproteins, such as podoplanin. METHODS: This study aimed to evaluate the participation of moesin and podoplanin in the invasive tumor front of oral squamous cell carcinomas, and their influence on patients' prognosis. Podoplanin and moesin immunoexpressions were evaluated by a semi-quantitative score method, based on the capture of 10 microscopic fields, at 400X magnification, in the invasive tumor front of oral squamous cell carcinomas. The association of moesin and podoplanin expression with clinicopathological variables was analyzed by the chi-square, or Fisher's exact test. The 5 and 10 years survival rates were calculated by the Kaplan-Meier method and the survival curves were compared by using the log-rank test. RESULTS: The immunohistochemical expression of moesin in the invasive front of oral squamous cell carcinomas was predominantly strong, homogenously distributed on the membrane and in the cytoplasm of tumor cells. The expression of moesin was not associated with clinical, demographic and microscopic features of the patients. Otherwise, podoplanin expression by malignant epithelial cells was predominantly strong and significantly associated with radiotherapy (p = 0.004), muscular invasion (p = 0.006) and lymph node involvement (p = 0.013). Strong moesin expression was considered an unfavorable prognostic factor for patients with oral squamous cell carcinomas, clinical stage II and III (p = 0.024). CONCLUSIONS: These results suggested that strong moesin expression by malignant cells may help to determine patients with oral squamous cell carcinoma and poor prognosis.

Podoplanin, ezrin, and Rho-A proteins may have joint participation in tumor invasion of lip cancer.

INTRODUCTION: Podoplanin and ezrin connection through Rho-A phosphorylation have been suggested as part of the activation pathway, in the process of tumor invasion and cell movement in oral squamous cell carcinomas. OBJECTIVE: The aim of this study was to evaluate the correlation among podoplanin, ezrin, and Rho-A immunoexpressions in 91 squamous cells carcinomas of the lower lip and their influence in patient's prognosis. MATERIAL AND METHODS: The immunoexpressions of podoplanin, ezrin, and Rho-A were evaluated through a semi-quantitative score method, based on the capture of 10 microscopic fields at the front of tumor invasion. The association and correlation of these proteins with the clinicopathological features were verified by Fischer's exact test and Spearman's test. The prognostic values were analyzed by Kaplan-Meier method and log-rank test. RESULTS: A statistically significant association between strong cytoplasmic podoplanin expression and alcohol (p = 0.024), loco-regional recurrences (p = 0.028), and lymph node metastasis (pN+) (p = 0.010) was found. The membranous (p = 0.000 and r = 0.384) and cytoplasmic (p = 0.000 and r = 0.344) podoplanin expression was statistically correlated with ezrin expression. Also, membranous podoplanin was significantly correlated with Rho-A expression (p = 0.006 and r = 0.282). The expressions of podoplanin, ezrin, and Rho-A were not significant prognostic factors for patients with squamous cell carcinomas of the lower lip. CONCLUSIONS: Therefore, our results confirm a correlation among podoplanin, ezrin, and Rho-A expressions in squamous cell carcinoma of the lip suggesting a cooperative participation of these proteins in cell movement and invasion. CLINICAL RELEVANCE: Furthermore, strong cytoplasmic podoplanin expression could be helpful to identify patients with squamous cell carcinoma of the lip and lower risk of loco-regional recurrences.

An Uncommon Occurrence of Three-Fourth Molars Concomitant to Hypodontia in a Nonsyndromic Patient.

Bilaterally mandibular and maxillary fourth molars, concomitant to hypodontia, are uncommon. These dental anomalies, called hypohyperdontia, are rare and etiology is not well defined. Several complications may occur as a consequence of supernumerary teeth, such as teeth impaction, resorption of adjacent teeth, and development of cysts and tumors. The authors present an uncommon occurrence of three-fourth molars concomitant to hypodontia and dentigerous cyst associated with impacted mandibular second and third molars in a nonsyndromic patient. The authors attempt to discuss the occurrence of bilaterally fourth molars and reinforce the importance of clinical approach, considering risks and benefits for the patient.

The relationship between ezrin and podoplanin expressions in keratocystic odontogenic tumors.

BACKGROUND: The aims of this study were to investigate the immunolocalization of ezrin and its relationship with the podoplanin expression in keratocystic odontogenic tumors. MATERIAL AND METHODS: The immunohistochemical expressions of ezrin and podoplanin by odontogenic epithelium were evaluated in keratocystic odontogenic tumors using monoclonal antibodies. RESULTS: Our results showed strong cytoplasmic ezrin and membranous podoplanin expressions in basal epithelial layer of all keratocystic odontogenic tumors. The cytoplasmic and membranous ezrin expressions were also detected in suprabasal epithelial layers of tumors. Statistically significant difference between cellular immunolocalization of ezrin and podoplanin odontogenic epithelium were found by Wilcoxon's test (p < 0.05). No correlation between both proteins in keratocystic odontogenic tumors was detected by Spearman test. CONCLUSIONS: These results suggest that ezrin and podoplanin may contribute to the expansive growth and local invasiveness of keratocystic odontogenic tumors. Additionally, as both proteins were overexpressed by odontogenic epithelium, their possible roles need to be further explored in benign odontogenic tumors.

Palisaded encapsulated neuroma in tongue - A commonly misdiagnosed peripheral nerve sheath tumor.

Palisaded encapsulated (solitary circumscribed) neuromas are benign neural tumors, rarely found in oral mucosa. This case reports a 24-year-old man that presented a unique soft nodule at the left side of the tongue. An excisional biopsy was performed under local anesthesia and histopathological examination of the surgical specimen revealed a well circumscribed mass composed of spindled Schwann cells, often aligned and fasciculated, forming occasional nodules embedded in a fibrous stroma. Histopathological analysis showed the presence of positive cells for S-100, EMA, CD57, and collagen IV. Based on clinical and microscopical features, the diagnosis established was of palisaded encapsulated neuroma. This case report aims to discuss the differential diagnosis among palisaded encapsulated neuroma and the other neural tumors that affect the oral mucosa.

A Rare Case of Intraductal Papilloma Arising from Minor Salivary Gland in the Floor of the Mouth.

A 77-year-old woman with a rare oral intraductal papilloma arising from the minor salivary gland located on the floor of the mouth and causing the mucus retention is reported. Microscopically, the lesion was characterized by unicystic cavity exhibiting the lumen partially filled by papillary projections of the ductal epithelium with varying degree of oncocytic metaplasia. Based on the histopathological analysis, the differential diagnosis of oral intraductal papillomas and other ductal neoplasms of salivary origin are discussed.

Beta adrenergic receptor activation inhibits oral cancer migration and invasiveness.

OBJECTIVE: The aim of this study was to verify ß2-AR expression in oral squamous cell carcinoma cell lines (SCC-9 and SCC-25), and to investigate the role of this receptor in migration and invasion of these neoplastic cells. DESIGN: SCC-9 and SCC-25 cells were investigated for gene and protein expression of ß2-AR. Cell migration and invasion were analyzed by wound healing assay and transwell invasion camera system. Different concentrations (0.1, 1 and 10 µM) of norepinephrine were used to stimulate, and 1 µM propranolol was used to block the beta-adrenergic receptors on cancer cells. Differences in median values of SCC-9 and SCC-25 and ß2-AR protein expression were analyzed by Friedman test and in case of significant differences; pairwise comparisons were performed using Bonferroni correction. RESULTS: The results showed that the ß2-AR gene and protein expression were observed in both oral cancer cell lines. The concentration of 10 µM of norepinephrine significantly inhibited (p ≤ 0.05) migration of SCC-9 and SCC-25 cell lines. Furthermore, there was a significant reduction (p ≤ 0.05) in the effect of norepinephrine on cell migration when the ß2-AR was inhibited by propranolol. The blockade by propranolol showed a tendency to reverse the effect of norepinephrine on the invasiveness of SCC-9 and SCC-25. CONCLUSIONS: The use of beta-adrenergic receptor agonists could become an adjuvant therapeutic target in the treatment of this malignancy.

Tissue eosinophilia and its association with tumoral invasion of oral cancer.

This study investigated if tumor-associated tissue eosinophilia (TATE) could be associated with the process of tissue invasion in oral squamous cell carcinomas (OSCCs) and its influence on patient's prognosis. Forty-three patients treated for OSCCs with or without lymph nodes involvement, at A. C. Camargo Cancer Hospital, Brazil, were selected for TATE analysis. Two degrees of tissue eosinophilia were established in OSCC: absent/mild and intense. The TATE was evaluated in relation to the clinicopathological features and prognostic value using chi( 2) test and the Kaplan-Meier method. Most of the patients with OSCC in advanced clinical stage presented muscular infiltration and significantly intense TATE whereas those with tumors in early stage frequently showed absent/mild eosinophilia (P = .009). The TATE showed no prognostic value for 5-year and 10-year survival rates of the OSCC. These findings suggest that intense TATE seems to reflect the stromal invasion of the OSCCs that occur in advanced clinical stage.

The stem cell markers expression CD44v6 and podoplanin in lip cancer: clinical significance.

This study aimed to analyze the immunoexpression of cancer stem cell markers, CD44v6, and podoplanin in 91 patients with lip squamous cell carcinomas (LSCC). The immunostaining of podoplanin and CD44v6 was evaluated in ten high-power fields (× 400 magnification) at the invasive front of LSCC, using a semi-quantitative score method. Chi-square test or Fisher's exact test was used to verify the association of podoplanin and CD44v6 expressions with clinicopathologic variables. Spearman's correlation test was used to analyze the correlation between the two antibodies in lip cancer. Disease-free survival probabilities in 5 and 10 years were estimated according to the Kaplan-Meier method and compared using the log-rank test. The independent effects of the significant variables were analyzed by Cox proportional hazards regression model. A strong podoplanin expression was observed in the membrane and cytoplasm of most lip tumor cells, and this was inversely associated with locoregional recurrence (p = 0.028) and with histopathological grade of malignancy (p = 0.026). Additionally, CD44v6 immunostaining was strongly expressed in the membrane of tumor cells in 95.4% of the LSCC. Patients with strong membranous (p = 0.016) or strong cytoplasmic (p = 0.030) podoplanin-positive tumors resulted in significantly better disease-free survival than those who had podoplanin weak/negative tumors, confirming podoplanin expression as a favorable independent prognostic factor. Podoplanin and CD44v6 were strongly expressed by tumor cells and podoplanin immunoexpression can help to determine lip cancer patients with lower risk for disease recurrence.

Atypical intraosseous mucoepidermoid carcinoma with two years of evolution and cutaneous infiltration.

The intraosseous mucoepidermoid carcinoma is a rare lesion that frequently affects the posterior region of the mandible. This case reports a multilocular osteolytic radiolucency with two years of evolution, that expanded and perforated the cortical bone, with irregular and indefinite margins, that extended from the mandibular angle to the ascendant ramus, with cutaneous ulceration, detected in a 51 years-old male. An incisional biopsy was performed and confirmed the diagnosis of low-grade intraosseous mucoepidermoid carcinoma. The patient was submitted to partial mandibulectomy, neck dissection and post-operative radiotherapy. In three years of follow-up, there was no evidence of tumor recurrence. This case report reinforces that even a low-grade intraosseous mandibular mucoepidermoid carcinoma tends to expand and to perforate the bone cortical, infiltrating to adjacent soft tissues, in long time of evolution. Key words:Intraosseous, mucoepidermoid carcinoma, mandible.

Immunoexpression of proteins involved in cytoskeleton remodeling in benign odontogenic lesions.

OBJECTIVE: The present study was designed to analyze the immunolocalization of proteins involved in cytoskeleton remodeling, such as moesin and Rho-A, in benign odontogenic lesions that present with expansive growth and invasive clinical behavior. MATERIALS AND METHODS: Expressions of moesin and Rho-A in odontogenic epithelium were evaluated by immunohistochemical analysis in 45 odontogenic lesions using monoclonal antibodies. RESULTS: Our results demonstrated strong membranous and cytoplasmic expressions of moesin in the epithelial cells in 66.7% and 44.4% of the odontogenic lesions, respectively. Furthermore, Rho-A expression in odontogenic epithelium was strong in the membrane and cytoplasm of 51.1% and 62.2% of the odontogenic lesions, respectively. A statistically significant correlation was found between the membranous and cytoplasmic expressions of moesin (p = 0.000) and those of Rho-A (p = 0.048) in odontogenic epithelial cells, while no statistically significant correlation was found between moesin and Rho-A expressions (p > 0.05). CONCLUSIONS: The present study confirmed the strong expressions of moesin and Rho-A by odontogenic epithelial cells, suggesting their involvement in the development of benign odontogenic lesions. However, this study has failed to detect the connection between the moesin and Rho-A interaction in expansive growth and local invasiveness of these lesions.

Moesin Involvement in Oral Carcinogenesis of the Lower Lip.

BACKGROUND/AIM: This study analyzed moesin immunoexpression in 91 lip squamous cell carcinomas and its influence in patients' prognosis. MATERIALS AND METHODS: Moesin immunoexpression was evaluated at the invasive tumor front by a semi-quantitative score method. The association of moesin with the clinicopathological variables was analyzed by the Chi-square test, the survival rates were calculated by Kaplan-Meier and the survival curves compared using the log-rank test. RESULTS: The expression of moesin was strong at the invasive tumor front and weak/negative in differentiated cells such as keratin pearls. There was no association between moesin expression and the clinicopathological variables, but there was a tendency for patients with lip cancer and strong moesin expression to have lower 5- and 10-year overall and disease-free survival rates. CONCLUSION: Our results confirm the participation of moesin in oral carcinogenesis and suggest that this protein can influence the survival rates of patients with lip squamous cell carcinoma.

A rare case report of an adenomatoid odontogenic tumor associated with odontoma in the maxilla.

INTRODUCTION: The adenomatoid odontogenic tumor (AOT) associated with odontoma has been described as a new entity with overlapping features of adenomatoid-like structures and odontoma. Presentation of the case: We report the second case of AOT associated with odontoma in the posterior maxillary region. A 22-year-old patient presented expansion of the vestibular and palatine cortical wall of maxilla. Radiographic examination and computed tomography confirmed erosion of maxillary walls, root resorption, displacement of a neighboring tooth and radiopacities within the lesion. Histopathological analysis showed a cystic cavity lined by odontogenic epithelium, organized as duct like structures and tubular dentin. The diagnosis established was AOT associated with odontoma. DISCUSSION: As fewer cases have been described, the aim of this study is to report clinical behavior and evolution of adenomatoid odontogenic tumor associated with odontoma, as it have not yet been described and its origin is not completely established. The hamartomatous or neoplastic origin of this tumor is under debate. CONCLUSIONS: Therefore, it is necessary similar cases to be published to increase the knowledge about the clinical behavior and evolution of this tumor, to enable such lesions to be more clearly defined in the next classification of odontogenic tumors.

Correlation of Beta-2 Adrenergic Receptor Expression in Tumor-Free Surgical Margin and at the Invasive Front of Oral Squamous Cell Carcinoma.

Background. The beta-2 adrenergic receptor is expressed by neoplastic cells and is correlated with a wide spectrum of tumor cell mechanisms including proliferation, apoptosis, angiogenesis, migration, and metastasis. Objectives. The present study aimed to analyze the expression of the beta-2 adrenergic receptor (ß2-AR) in tumor-free surgical margins of oral squamous cell carcinomas (OSCC) and at the invasive front. Sixty-two patients diagnosed with OSCC, confirmed by biopsy, were selected for the study. The clinicopathological data and clinical follow-up were obtained from medical records and their association with ß2-AR expression was verified by the chi-square test or Fischer's exact test. To verify the correlation of ß2-AR expression in tumor-free surgical margins and at the invasive front of OSCCs, Pearson's correlation coefficient test was applied. Results. The expression of ß2-AR presented a statistically significant correlation between the tumor-free surgical margins and the invasive front of OSCC (r = 0.383; p = 0.002). The immunohistochemical distribution of ß2-AR at the invasive front of OSCC was also statistically significant associated with alcohol (p = 0.038), simultaneous alcohol and tobacco consumption (p = 0.010), and T stage (p = 0.014). Conclusions. The correlation of ß2-AR expression in OSCC and tumor-free surgical margins suggests a role of this receptor in tumor progression and its expression in normal oral epithelium seems to be constitutive.

Correlation of Beta-2 Adrenergic Receptor Expression inTumor-Free Surgical Margin and at the Invasive Front ofOral Squamous Cell Carcinoma

Background. The beta-2 adrenergic receptor is expressed by neoplastic cells and is correlated with a wide spectrum of tumor cell mechanisms including proliferation, apoptosis, angiogenesis, migration, and metastasis. Objectives. The present study aimed to analyze the expression of the beta-2 adrenergic receptor (ß2-AR) in tumor-free surgical margins of oral squamous cell carcinomas (OSCC) and at the invasive front. Sixty-two patients diagnosed with OSCC, confirmed by biopsy, were selected for the study. The clinicopathological data and clinical follow-up were obtained from medical records and their association with ß2-AR expression was verified by the chi-square test or Fischer's exact test. To verify the correlation of ß2-AR expression in tumor-free surgical margins and at the invasive front of OSCCs, Pearson's correlation coefficient test was applied. Results. The expression of ß2-AR presented a statistically significant correlation between the tumor-free surgical margins and the invasive front of OSCC (r = 0.383; p = 0.002). The immunohistochemical distribution of ß2-AR at the invasive front of OSCC was also statistically significant associated with alcohol (p = 0.038), simultaneous alcohol and tobacco consumption (p = 0.010), and T stage (p = 0.014). Conclusions. The correlation of ß2-AR expression in OSCC and tumor-free surgical margins suggests a role of this receptor in tumor progression and its expression in normal oral epithelium seems to be constitutive