Grading Of Oral Cancer Research Articles

An integrated approach for identification of a panel of candidate genes arbitrated for invasion and metastasis in oral squamous cell carcinoma

Oral squamous cell carcinoma (OSCC) is known for its aggressiveness associated with poor prognosis. The molecular mechanisms underlying the invasion and metastasis are still poorly understood. An improved understanding of these mechanisms shall precede the development of new diagnostic tools and targeted therapies. We report an integrated approach using bioinformatics to predict candidate genes, coupled with proteomics and immunohistochemistry for validating their presence and involvement in OSCC pathways heralding invasion and metastasis. Four genes POSTN, TNC, CAV1 and FSCN1 were identified. A protein–protein interaction network analysis teamed with pathway analysis led us to propose the role of the identified genes in invasion and metastasis in OSCC. Further analyses of archived FFPE blocks of various grades of oral cancer was carried out using TMT-based mass spectrometry and immunohistochemistry. Results of this study expressed a strong communiqué and interrelationship between these candidate genes. This study emphasizes the significance of a molecular biomarker panel as a diagnostic tool and its correlation with the invasion and metastatic pathway of OSCC. An insight into the probable association of CAF's and these biomarkers in the evolution and malignant transformation of OSCC further magnifies the molecular-biological spectrum of OSCC tumour microenvironment.

Iron expression in the biopsies of patients with different grades of Oral Squamous Cell Carcinoma

Oral Squamous Cell Carcinoma (OSCC) is becoming a frequent problem all over the world and its etiological factors are different in different areas of the world. Iron is a pivotal nutrient and its deficiency is considered as malnutrition. The possibility that low iron intake might cause oral cancer needs to be investigated Objective: To evaluate the expression of iron in different grades of oral cancer. Methods: A total of 40 biopsy samples of OSCC were included in this study, comprising of 35% well and moderately differentiated and 25% in poorly differentiated cases.Iron staining was performed on all the cases and routine hematoxylin-eosin staining was also performed. Cases were then assessed by two independent hisptopathologists Results: Increased expression of iron in early stage of cancer was observed, which is well differentiated stage. Moderate expression was observed in moderately differentiated stage and almost no expression was observed in highly advanced stage of cancer which is poorly differentiated stage. Conclusions: According to this study, it has been concluded that there is more prevalence of oral cancer in males than in females because the males of this region are more involve in cigratte smoking and the use of oral drugs like pan, gutka, mishri and other types of chewing drugs. Iron deficiency in the body may lead to OSCC.

Quantitative analysis of tumor-associated tissue eosinophils and tumor-associated blood eosinophils in oral squamous cell carcinoma.

Background:Stromal response to cancer is usually characterized by intense lymphoplasmacytic infiltrate. However, recently, the attention has shifted to tumor-associated tissue eosinophils (TATE). Tumor-associated blood eosinophils (TABE) are rare in solid cancers; however, carcinoma of the head and neck shows its prevalence.Aim:The aim of the study was to investigate the prevalence and relationship of tissue and blood eosinophils in various grades of oral cancer. The purpose of the article is to emphasize the possible clinical and biological significance of eosinophils in patients of oral squamous cell carcinoma (OSCC) so that appropriate therapeutic strategies can be devised accordingly.Study Design:Thirty histologically confirmed cases of oral squamous cell carcinoma were divided into well, moderate and poorly differentiated carcinoma. Eosinophilic infiltration in the tissue was graded as low, moderate and massive TATE. The number of eosinophils per 100 WBCs was taken as the differential eosinophil count. Blood eosinophilia (BE) >6% was considered to be TABE.Materials and Methods:Hematoxylin and eosin-stained tissue sections at 5 μ were evaluated. Prolonged staining in dilute 0.05% aqueous eosin demonstrated eosinophils selectively. Blood smears were stained by Leishman stain.Statistical Analysis:Student's t-test, Chi-square test, ANOVA, Newman–Keuls Multiple Comparison Test and Karl Pearson correlation coefficient® method were used.Results:The mean TATE value was highest in poorly differentiated carcinoma. TABE was seen only in a few cases and was associated mostly with poorly differentiated OSCC.Conclusion:There was a statistically significant correlation between TATE and histological grades of OSCC. Eosinophilia of the peripheral blood is an adverse sign in patients with carcinoma.

A Novel Surface Enhanced Raman Catheter for Rapid Detection, Classification, and Grading of Oral Cancer.

Fabrication and testing of a novel nanostructured surface-enhanced Raman catheter device is reported for rapid detection, classification, and grading of normal, premalignant, and malignant tissues with high sensitivity and accuracy. The sensor part of catheter is formed by a surface-enhanced Raman scattering (SERS) substrate made up of leaf-like TiO2 nanostructures decorated with 30 nm sized Ag nanoparticles. The device is tested using a total of 37 patient samples wherein SERS signatures of oral tissues consisting of malignant oral squamous cell carcinoma (OSCC), verrucous carcinoma, premalignant leukoplakia, and disease-free conditions are detected and classified with an accuracy of 97.24% within a short detection-cum-processing time of nearly 25-30 min per patient. Neoplastic grade changes detected using this device correlate strongly with conventional pathological data, enabling correct classification of tumors into three grades with an accuracy of 97.84% in OSCC. Thus, the potential of a SERS catheter device as a point-of-care pathological tool is shown for the rapid and accurate detection, classification, and grading of solid tumors.

Cancer stem cell markers in patterning differentiation and in prognosis of oral squamous cell carcinoma.

Differentiation is a major histological parameter determining tumor aggressiveness and prognosis of the patient; cancer stem cells with their slow dividing and undifferentiated nature might be one of the factors determining the same. This study aims to correlate cancer stem cell markers (CD44 and CD147) with tumor differentiation and evaluate their subsequent effect on prognosis. Immunohistochemical analysis in treatment naïve oral cancer patients (n = 53) indicated that the expression of CD147 was associated with poorly differentiated squamous cell carcinoma and moderately differentiated squamous cell carcinoma (p < 0.01). Furthermore, co-expression analysis showed that 45% each of moderately differentiated squamous cell carcinoma and poorly differentiated squamous cell carcinoma patients were CD44high/CD147high as compared to only 10% of patients with well-differentiated squamous cell carcinoma. A three-way analysis indicated that differentiation correlated with recurrence and survival (p < 0.05) in only the patients with CD44high/CD147high cohort. Subsequently, relevance of these cancer stem cell markers in patterning the differentiation characteristics was evaluated in oral squamous cell carcinoma cell lines originating from different grades of oral cancer. Flowcytometry-based analysis indicated an increase in CD44+/CD147+ cells in cell lines of poorly differentiated squamous cell carcinoma (94.35 ± 1.14%, p < 0.001) and moderately differentiated squamous cell carcinoma origin (93.49 ± 0.47%, p < 0.001) as compared to cell line of well-differentiated squamous cell carcinoma origin (23.12% ± 0.49%). Expression profiling indicated higher expression of cancer stem cell and epithelial-mesenchymal transition markers in SCC029B (poorly differentiated squamous cell carcinoma originated; p ≤ 0.001), which was further translated into increased spheroid formation, migration, and invasion (p < 0.001) as compared to cell line of well-differentiated squamous cell carcinoma origin. This study suggests that CD44 and CD147 together improve the prognostic efficacy of tumor differentiation; in vitro results further point out that these markers might be determinant of differentiation characteristics, imparting properties of increased self-renewal, migration, and invasion.

Cyclooxygenase-2 expression in oral precancerous and cancerous conditions and its inhibition by caffeic acid phenyl ester-enriched propolis in human oral epidermal carcinoma KB cells

Oral cancer accounts for 3-5% of all cancers worldwide. The present study was undertaken to investigate the correlation between overexpression of cyclooxygenase-2 (COX-2) and various grades of oral cancer, and to ascertain the inhibitory effect of propolis in the human oral carcinoma cell line. For ex vivo studies, 45 patients with oral submucous fibrosis (OSF; n=15), oral leukoplakia (OLP; n=18) and oral squamous cell carcinoma (OSCC; n=18) were recruited, and a biopsy was done to determine COX-2 protein expression by Western blotting and immunohistochemistry (IHC). For the in vitro study, COX-2 levels were measured in human oral epidermal carcinoma cell line by immune blotting and IHC. The results of ex vivo studies by Western blotting revealed that COX-2 protein levels were highly upregulated in OSCC tissue, followed by OLP and OSF. The levels of COX-2 expression also showed a positive correlation with the grade (severity) of each oral precancerous and cancerous condition. Immunohistochemistry analysis revealed the presence of intense COX-2 staining in the cells of OSCC tissue, equivalent to the OLP and OSF specimens. In the in vitro study of oral carcinoma KB cells, Western blotting and IHC analysis showed that caffeic acid phenyl ester (CAPE)-rich propolis and celecoxib, a standard COX-2 inhibitor, markedly downregulated COX-2 expression. These results suggest that propolis exhibits a chemopreventive potential by lowering COX-2 expression in the oral carcinoma KB cell line. Hence, propolis might be used as an adjuvant therapy for treating oral cancer with standard chemotherapy drugs.

Expression of type III collagen correlates with poor prognosis in oral squamous cell carcinoma.

The aim of the present study was to evaluate, compare, and correlate the types of collagen fibers seen in different grades of oral cancer. Thirty cases of histologically-diagnosed, well-, moderately-, and poorly-differentiated oral squamous cell carcinoma (OSCC) were retrieved from the archives of the Institute. Collagen was evaluated using picrosirius red stain and immunohistochemical analysis of the antibody to type III collagen. A correlation between these findings and the grade of OSCC was evaluated. Collagen fibers showed a change in birefringence ranging from reddish-orange to greenish-yellow in well- to poorly-differentiated oral squamous cell carcinoma. The findings were statistically significant for polarizing colors observed in grades of OSCC (P<.001). Immunohistochemical staining intensity of type III collagen changed from weak to strong as grade increased for OSCC, and was also statistically significant (P<.001). In the present study, tumor progression reflected a change in collagen present, from type I to type III. Determination of the type of collagen in different grades of OSCC can facilitate therapeutic targeting of molecules responsible for invasion and progression of oral cancer.

Fucose: A biomarker in grading of oral cancer.

Introduction:Early diagnosis of cancer helps a great deal in the management of oral cancer patients. Number of proteinous markers have been employed for this purpose. Majority of them are not specific. Recently conjugated oligosaccharide with proteins and lipids have gained considerable importance in the present postgenomics and postproteomic period in the diagnostic and prognostics of cancer cases.Materials and Methods:In this study, serum fucose levels were estimated in 50 control cases and 75 cases of oral cancer by the method of Dische and Shettles as adopted by Winzler.Results:Serum fucose levels were found to be significantly higher in oral cancer cases (46.63 ± 5.29 mg/dl) as compared to the control cases (7.22 ± 0.26 mg/dl). The stepwise elevated serum fucose levels were found to be correlated with the histopathological grading of oral cancer.Conclusions:Estimation of such fucose conjugated proteins is suggestive to be good biomarkers in the diagnosis of oral cancer cases as well as in assessing the prognosis of such cases.

Laser‐induced autofluorescence spectral ratio reference standard for early discrimination of oral cancer

Laser-induced autofluorescence (LIAF) is an emerging noninvasive technique in the biomedical field, especially for cancer detection. The goal of the study was to develop a spectral ratio reference standard (SRRS) to discriminate different grades of oral cancer. LIAF emission spectra from oral mucosa were recorded in the 420-720 nm spectral range on a miniature fiberoptic spectrometer from 14 anatomical sites of 35 healthy volunteers and 91 sites of 44 patients, with excitation at 404 nm from a diode laser. Histopathologic analysis of biopsy samples showed that oral mucosa of adjoining malignant sites in patients are not usually normal, but showed various degrees of epithelial dysplasia and hyperplasia. Therefore, instead of using LIAF data from apparently normal lesions of patients as control, spectral data values of the oral mucosa of healthy volunteers were used as control. The autofluorescence emission at 500 nm is characteristic of oral mucosa, whereas in malignant lesions a new peak is seen at 685 nm in addition to the previously reported peaks at 635 and 705 nm. Three spectral ratio reference standard (SRRS) scatterplots were created to differentiate the normal mucosa from hyperplasia, hyperplasia from dysplasia, and dysplasia from squamous cell carcinoma (SCC) using the mean fluorescence intensity ratios (F500/F635, F500/705 and F500/F685) measured from 40 sites in 20 patients and 11 sites in 35 healthy volunteers. During blind tests at 21 sites in 17 patients all 3 SRRS plots showed 100% sensitivity and specificity to discriminate hyperplasia from dysplastic and normal tissues, whereas only the F500/F685 SRRS showed the same sensitivity and specificity to differentiate dysplasia from SCC. An SRRS criteria based on scatterplots of autofluorescence spectral intensity ratios is described to discriminate oral mucosal variations and screen early stages of tissue progression toward malignancy.

Oxygenated hemoglobin diffuse reflectance ratio for in vivo detection of oral pre-cancer

Diffuse reflectance (DR) spectroscopy is a simple, low-cost, and noninvasive modality with potential for distinguishing oral precancer. Recently, in an ex vivo study, the DR spectral ratio (R545/R575) of oxygenated hemoglobin bands at 545 and 575 nm was used for grading malignancy. This work presents the results of clinical trials conducted in 29 patients to detect oral precancer using this ratio. We use site-specific normal spectra from a group of 36 healthy volunteers for comparison with those of patients. Toward this, in vivo DR spectra from 14 anatomical sites of the oral cavity of healthy volunteers are recorded on a miniature fiber optic spectrometer with white light excitation. The R545/R575 ratio is lowest for healthy tissues and appears to increase with the grade of malignancy. As compared to scatter plots that use the mean DR ratio from all anatomical sites, those using site-specific data show improved sensitivity and specificity for early diagnosis and grading of oral cancer. In the case of buccal mucosa, using scatter plots of R545/R575 ratio, we obtain a sensitivity of 100% and specificity of 86% for discriminating precancer (dysplasia) from hyperplasia, and a sensitivity of 97% and specificity of 86% for discriminating hyperplasia from normal.