CD34 Angiogenesis Marker in Oral Squamous Cell Carcinoma: An Immunohistochemical Study.

A histopathologic comparison between synchronous and single primary oral squamous cell carcinomas

PurposeThe aim of the study is to investigate the immunohistochemical expression of both Alpha smooth muscle actin and Transforming Growth Factor beta and compare their expression in oral papillary squamous cell carcinoma with their expression in different histological grades of oral squamous cell carcinoma. A correlation between these immuno-histochemical expressions and histological findings will then be performed. The research question is “Do the percentages of α-SMA and TGF-β immune-expression in OPSCC differ from that in the conventional OSCC?”.MethodsThis will be achieved by collecting archival blocks of oral papillary squamous cell carcinoma and different grades of oral squamous cell carcinoma, staining the specimens with Transforming Growth Factor beta and alpha smooth muscle actin, then measuring the mean staining index of expression in each group and the area percent of both markers.ResultsResults revealed that transforming growth factor beta expression in the epithelium was high in all cases of well-differentiated squamous cell carcinoma, most oral papillary squamous cell carcinoma, and poorly differentiated oral squamous cell carcinoma. On the other hand, different grades of oral squamous cell carcinoma showed a high staining index of alpha smooth muscle actin expression in the stroma. While cases of oral papillary squamous cell carcinoma were either moderate or low-staining.ConclusionsOral papillary squamous cell carcinoma has a favourable prognosis compared to different histological grades, and the prognosis does not depend only on histological grade but also on other prognostic factors.

MicroRNA-based classifiers for diagnosis of oral cavity squamous cell carcinoma in tissue and plasma

Background:Oral squamous cell carcinoma (OSCC) is the most prevalent carcinoma of the head-and-neck region. Despite advanced tissue diagnosis, the survival period remains at 5 years. New molecular and cell markers are under investigation to improve survival rate.Objective:The objective of this study was to assess angiogenic activity by analyzing CD105 microvascular density (MVD) count in different histologic grades of OSCC.Subjects and Methods:Sixty samples are categorized into four groups. Immunohistochemical method with anti-CD105 was used for analyzing MVD.Results:The mean count of MVD is higher in OSCC than in normal mucosa and also significant among different histologic grades. A highly significant correlation is observed among different groups of OSCC samples.Conclusion:CD105 is a predictive marker for neoangiogenesis in OSCC.

The TNM staging system for oral squamous cell carcinoma (OSCC) provides clinicians a dependable foundation for patient prognosis and management decisions, but in clinical practice, treatment outcomes of patients with OSCC are sometimes unsatisfactory. This retrospective study investigated the association between survival and clinicopathological characteristics and histological grades of 2535 patients with OSCC. Additionally, the present study aimed to compare the predictive abilities of histological grades with other common prognostic factors. The enrolled patients were divided into three groups by two experienced pathologists into well-differentiated, moderately differentiated, and poorly differentiated groups, according to the WHO classification. Finally, we designed an observational, retrospective study based on the histological grading of tumors to compare their clinicopathological characteristics and conducted survival analysis among the three groups. Advanced tumor stage was diagnosed in 23.9%, 44.0%, and 55.1% of patients with grades 1-3 OSCC, respectively. By T status, T3 or T4 tumors were found in approximately 22%, 34%, and 40% of patients with grades 1-3 OSCC, respectively. By N status, lymph node metastases were found in 6.1%, 29.3%, and 45.9% of patients with grades 1-3 OSCC, respectively. Thus, significant survival differences were observed based on different OSCC histological grades. Meanwhile, in the multivariate (adjusted) analysis, N1 and N2 stages, extranodal spread, and poor differentiation were associated with a higher recurrence risk than the other common prognostic factors. In conclusion, 5% of patients in our study presented with poorly differentiated OSCC at diagnosis. Furthermore, grade 3 OSCC has worse prognosis and is more aggressive than grades 1 and 2 OSCC. In the future, we should focus on modifying individual therapy for poorly differentiated OSCC to achieve improved outcomes.

The TNM staging system for oral squamous cell carcinoma (OSCC) provides clinicians a dependable foundation for patient prognosis and management decisions, but in clinical practice, treatment outcomes of patients with OSCC are sometimes unsatisfactory. This retrospective study investigated the association between survival and clinicopathological characteristics and histological grades of 2535 patients with OSCC. Additionally, the present study aimed to compare the predictive abilities of histological grades with other common prognostic factors. The enrolled patients were divided into three groups by two experienced pathologists into well-differentiated, moderately differentiated, and poorly differentiated groups, according to the WHO classification. Finally, we designed an observational, retrospective study based on the histological grading of tumors to compare their clinicopathological characteristics and conducted survival analysis among the three groups. Advanced tumor stage was diagnosed in 23.9%, 44.0%, and 55.1% of patients with grades 1–3 OSCC, respectively. By T status, T3 or T4 tumors were found in approximately 22%, 34%, and 40% of patients with grades 1–3 OSCC, respectively. By N status, lymph node metastases were found in 6.1%, 29.3%, and 45.9% of patients with grades 1–3 OSCC, respectively. Thus, significant survival differences were observed based on different OSCC histological grades. Meanwhile, in the multivariate (adjusted) analysis, N1 and N2 stages, extranodal spread, and poor differentiation were associated with a higher recurrence risk than the other common prognostic factors. In conclusion, 5% of patients in our study presented with poorly differentiated OSCC at diagnosis. Furthermore, grade 3 OSCC has worse prognosis and is more aggressive than grades 1 and 2 OSCC. In the future, we should focus on modifying individual therapy for poorly differentiated OSCC to achieve improved outcomes.

Expression of KITENIN and its association with tumor progression in oral squamous cell carcinoma

Immunohistochemical evaluation of Yes-associated protein1 (YAP1) expression in oral leukoplakia and squamous cell carcinoma: Implications for prognosis.

Angiogenesis is a complex event mediated by angiogenic factors released from cancer cells and immune cells. It has been reported to be associated with progression, aggressiveness and metastases of various malignant tumors including oral squamous cell carcinoma (OSCC). Similarly, mast cells have also been reported to play a role in tumor progression and metastases by promoting angiogenesis. The present study aims at comparison of microvascular density (MVD) and mast cell density (MCD) in normal oral mucosa (NM) and among various grades of OSCC. MVD was assessed immunohistochemically using anti-Factor VIII related von Willebrand factor, and MCD using anti-mast cell tryptase in a study sample of 30 cases of OSCC and 10 cases of clinically normal oral mucosa. The mast cells in normal oral mucosa and oral squamous cell carcinoma strongly expressed mast cell tryptase. The density of mast cells and micro vessels were significantly higher in OSCC compared to normal oral mucosa. The MCD and MVD were higher in moderately differentiated OSCC than in well differentiated OSCC ( P > 0.05) and normal oral mucosa ( P < 0.05). Pearson's correlation revealed a positive correlation between MCD and MVD ( r=0.33; P=0.077). These findings indicate that mast cells may play a role in up regulation of tumor angiogenesis in OSCC probably through mast cell tryptase.

Background. Oral squamous cell carcinoma is the sixth most frequent malignant tumor worldwide and the third most common cancers in developing countries. Oral leukoplakia is the best-known precursor lesion of oral squamous cell carcinoma. The aim of the present study was to compare immunohistochemical expression of antiapoptotic protein survivin in normal oral mucosa, oral leukoplakia, and oral squamous cell carcinoma. Method. Total 45 specimens of formalin fixed paraffin embedded tissue blocks, 15 in each of the following: normal oral mucosa, leukoplakia, and oral squamous cell carcinoma were used for the study. Immunohistochemical reaction for survivin protein was performed for the 4 µm thick histological sections taken on positively charged slides. Results. 20% normal mucosa cases, 53.33% cases of leukoplakia, and 80% of oral squamous cell carcinoma were found out to be survivin positive. One way ANOVA test indicated statistically significant difference of survivin expression between the three different groups (p < 0.001). Conclusion. A high incidence of survivin protein expression in oral epithelial dysplasia and squamous cell carcinoma samples indicate that survivin protein expression may be an early event in initiation and progression of oral squamous cell carcinoma.

Background:Oral squamous cell carcinoma (OSCC) is the most common malignancy in this region, and thus, further elucidation of its tumoral mechanisms is important. One of the main roles of the acute-phase protein orosomucoid-1 (ORM1) is the promotion of angiogenesis, which is key for tumor nutrition and growth.Aim:Our aim was to evaluate the immunohistochemical expression of ORM1 and the angiogenic activity indicated by microvascular density (MVD) in OSCC samples according to histological grade.Materials and Methods:Formalin-fixed, paraffin-embedded sections from 45 OSCC cases were submitted to immunohistochemistry: 25 were well-differentiated OSCC, 18 were moderately differentiated OSCC and 2 were poorly differentiated OSCC. ORM1 staining was evaluated by a semiquantitative method, and CD34-positive blood vessels were quantified to calculate the MVD. The results were statically analyzed.Results:All cases exhibited immunoexpression of ORM1 and CD34. However, no significant differences were found between the expression of both markers among the histological grades. In addition, the presence of ORM1 in inflammatory cells and in the extracellular matrix was detected in most cases.Conclusion:These results suggest that the induction of angiogenesis is not the main role of ORM1 in OSCC and may be associated with the regulation of the immune/inflammatory response or the transport of protumoral molecules, such as sialyl-Lewis X or phorbol esters, which requires confirmation in future studies.

Toll-like receptors (TLRs) play an essential role in the activation of innate immunity. TLRs are expressed in B-lymphocytes, monocytes, dendritic cells and epithelial cells. We examined the immunohistochemical expressions of TLR4 and TLR9 in various grades of oral epithelial dysplasia (OED), oral squamous cell carcinoma (OSCC) and normal oral mucosa (NOM) to determine the association between TLR4 and TLR9 in the progression of lesions from dysplasia to carcinoma. Expressions of TLR4 and TLR9 were assessed using immunohistochemistry (IHC) on paraffin embedded tissue blocks of various grades of OED (28 cases), OSCC (27 cases) and NOM (10 cases). Expression of TLR4 was high in all grades of OED and OSCC. Expression of TLR9 was high in well differentiated squamous cell carcinoma and moderately differentiated squamous cell carcinomas, and moderate to low in poorly differentiated squamous cell carcinomas. Although expression was high in case of TLR4, it was not statistically significant. Expression of TLR9 was statistically significant. In OED, expression of TLR9 was less than that of TLR4. Our results indicated that the pattern of expression of TLR4 and TLR9 increased significantly from mild to severe dysplasia compared to controls. Expression of TLR4 and TLR9 reflects progression of OED to OSCC, which suggests that TLR may play a role in tumorigenesis and that it could be used as a target for OSCC prevention and therapy in the future.

Studies have addressed the relevance of c-Jun and c-Fos proteins in cancer development. In the present study, the expression of c-Jun and c-Fos, the major components of transcription factor activator protein (AP1), were evaluated to determine possible alterations to these factors in oral squamous cell carcinoma (OSCC). Fifteen cases of normal oral mucosa and 20 cases of OSCC were retrieved from the Archives of the Surgical Pathology Service at the University of São Paulo. The samples of normal oral mucosa or OSCC originated from different oral mucosal sites. Tissues were submitted for immunohistochemical analysis to detect c-Jun and c-Fos proteins. The OSCC was classified as well, intermediate or poorly differentiated. The results showed that both c-Jun and c-Fos are expressed in normal oral mucosa and in OSCC. In normal mucosa, immunoreactivity for c-Jun was detected in the cytoplasm of the upper basal layers, while in OSCC, c-Jun was detected in the nuclei of the cells. C-Fos expression was observed in the nuclei of cells, both in normal mucosa and in OSCC, but its expression varied according to the cell layer in normal mucosa, and the differentiation of OSCC. The nuclear expression of c-Jun in OSCC, in contrast to its cytoplasmic expression in normal oral mucosa, indicates that c-Jun may have a role in the development of oral cancer. In contrast, the absence of both c-Jun and c-Fos in poorly differentiated carcinoma might be useful in understanding the cell cycle events important in uncontrolled cell growth.

Cdk4 and cdk6, key players in G1 phase, have been shown to play an important role in the development of oral squamous cell carcinoma (OSCC). This study investigated the expression of these two proteins in OSCC and premalignant lesions including oral leukoplakia (OL) with and without dysplasia and determined if alterations in the expression of these two proteins could be used as markers of malignant transformation. Expressions of cdk4 and cdk6 were evaluated in 61 samples including OSCC, OL with and without dysplasia and normal oral mucosa using immunohistochemistry method. Nuclear staining of the keratinocytes was considered positive and the percentage of positive cells was calculated. Expression of cdk4 was found in 11/15 (73.33%) OSCC, 13/14 (92.85%) OL with dysplasia, 13/20 (65%) OL without dysplasia and 3/12 (25%) normal mucosa. Expression of cdk6 was detected in 9/15 (60%) OSCC, 3/14 (21.43%) OL with dysplasia, 5/20 (25%) OL without dysplasia and 1/12 (8.33%) normal mucosa. In cdk4 stained specimens, the frequency of positive cases and the percentage of positive cells in normal mucosa was significantly lower than OL with dysplasia and OSCC. For cdk6 staining, the prevalence of positive cases and the percentage of positive cells in normal mucosa were significantly lower than OSCC. Overexpressions of cdk4 and cdk6 were observed in OSCC, indicating that these two proteins play a crucial role in OSCC. The aberrant expression of cdk4 was found in OL with dysplasia, suggesting that cdk4 may be involved in the early event of carcinogenesis.

Background Oral squamous cell carcinoma (OSCC) is the most prevalent malignant head and neck tumor, excluding the nonmelanoma skin cancer. Despite recent advances in the diagnosis and treatment, the disease's mortality rate is nonetheless high. The presence of isolated neoplastic cells or small clusters of up to four cells at the tumor’s invasive front, named tumor budding, is associated with a worse prognosis in OSCC. Angiogenesis has also been recognized as a determining factor in the progression of malignancies and in the development of metastases. Several studies have investigated the assessment of microvascular density (MVD) as a potential prognostic factor in OSCC. This study aimed to evaluate, in OSCC, differences in MVD between tumors with high-intensity tumor budding and tumors with low-intensity or no tumor budding. In samples with high-intensity tumor budding, differences in MVD between the budding area and the area outside the budding were also evaluated. Moreover, the study assessed differences in MVD concerning clinicopathological characteristics such as sex, age, tobacco smoking, tumor location and tumor size.Material and Methods One hundred and fifty [150] samples of OSCC were subjected to immunohistochemistry to assess the intensity of tumor budding (by immunostaining for multi-cytokeratin) and MVD (by immunostaining for CD34 and CD105, independently). The data were treated using descriptive and analytical statistics.Results There were no differences in MVD, assessed by immunostaining for CD34 or CD105, concerning clinicopathological characteristics such as sex, age, tobacco smoking, tumor location and tumor size (p > 0.05). Tumors with high-intensity tumor budding did not show differences in MVD, assessed by immunostaining for CD34 or CD105, when compared to tumors with low-intensity or no tumor budding (p > 0.05). However, in samples with high-intensity tumor budding, the MVD assessed by immunostaining for CD34 was higher in the budding area than in the area outside the budding (p < 0.05). This difference was not observed when MVD was assessed by immunostaining for CD105 (p > 0.05).Conclusions The higher MVD in the budding area may be an additional indication that this is a peculiar region of the tumor, associated with biological phenomena related to tumor progression. Key words:Oral squamous cell carcinoma, tumor budding, microvascular density.