Oral squamous cell carcinoma: a clinicopathological study on 194 cases in northeastern Brazil. A cross-sectional retrospective study

<div><p>ABSTRACT BACKGROUND: Only a few studies have evaluated the clinicopathological features of oral squamous cell carcinoma (SCC) in Brazil, and most were conducted in the most industrialized region of the country, i.e. the southeastern region. The aim of this study was to evaluate the clinicopathological features of this malignant neoplasm in northeastern Brazil. DESIGN AND SETTING: Retrospective study performed in an oral pathology laboratory in Recife, Brazil. METHODS: All cases of oral SCC that occurred between 2000 and 2015 were studied. Clinical data were recorded and histological slides were reviewed. Statistical analysis was performed using the chi-square test (P ≤ 0.05). RESULTS: A total of 194 cases were evaluated. The male-to-female ratio was 1.5:1. The mean age was 65.4 years, and only 6.6% of the cases occurred in patients younger than 41 years. Most tumors consisted of well-differentiated SCC (54.6%). CONCLUSIONS: The findings of this study highlight the higher prevalence of oral SCC among women and the increasing number of cases among young patients. Thus there is no specific risk group for oral SCC, as in the past. This fact needs to be taken into consideration in clinical routine care, so that apparently innocuous malignant lesions do not go unnoticed in these individuals.</p></div

Oral squamous cell carcinoma: a clinicopathological study on 194 cases in northeastern Brazil. A cross-sectional retrospective study

<div><p>ABSTRACT BACKGROUND: Only a few studies have evaluated the clinicopathological features of oral squamous cell carcinoma (SCC) in Brazil, and most were conducted in the most industrialized region of the country, i.e. the southeastern region. The aim of this study was to evaluate the clinicopathological features of this malignant neoplasm in northeastern Brazil. DESIGN AND SETTING: Retrospective study performed in an oral pathology laboratory in Recife, Brazil. METHODS: All cases of oral SCC that occurred between 2000 and 2015 were studied. Clinical data were recorded and histological slides were reviewed. Statistical analysis was performed using the chi-square test (P ≤ 0.05). RESULTS: A total of 194 cases were evaluated. The male-to-female ratio was 1.5:1. The mean age was 65.4 years, and only 6.6% of the cases occurred in patients younger than 41 years. Most tumors consisted of well-differentiated SCC (54.6%). CONCLUSIONS: The findings of this study highlight the higher prevalence of oral SCC among women and the increasing number of cases among young patients. Thus there is no specific risk group for oral SCC, as in the past. This fact needs to be taken into consideration in clinical routine care, so that apparently innocuous malignant lesions do not go unnoticed in these individuals.</p></div

Downregulation of activin A leads to a decrease in proliferation.

<p>Treatment with recombinant activin A was not able to promote proliferation of HaCat cells, as revealed by bromodeoxyuridine (BrdU) incorporation index (A) and cell cycle analysis (B). (C) Follistatin at 100 ng/ml significantly blocked BrdU incorporation in SCC-9 ZsGreen LN-1 cells, but no effects on cell cycle distribution were observed (D). Knockdown of activin A significantly decreased proliferation (E), enhancing the number of cells at G0/G1 and reducing the number in the S phase of cell cycle (F). Bars represent the means ± SD of three independent experiments. *p<0.01, **p<0.001, ***p<0.0001.</p

Expression of INHBA is inversely correlated with miR-143 and miR-145 expression in OSCC cells lines and tumor samples.

<p>(A) Spearman correlation analysis between the expressions of INHBA and miR-143 and miR-145 in 7 OSCC cell lines (A) and in 11 OSCC fresh samples (B). A significant inverse correlation was observed between both miR-143 and miR-145 and INHBA expression levels.</p

Spearman correlation between activin A immunohistochemical expression and clinicopathological variables.

<p>Spearman correlation between activin A immunohistochemical expression and clinicopathological variables.</p

Knockdown of activin A increases cyclin-dependent kinase inhibitors p16, p21 and p27.

<p>Cells were harvested, lysed and proteins were subjected to western blot analysis using specific antibodies against p16, p21, p27, CDK2, CDK4, CDK6, cyclin D1, cyclin E and phospho-RB. β-actin was used as loading control. A strong increase in expression of p16, p21 and p27, concomitant with decrease in phosphorylation of RB, was observed in shINHBA cells in comparison with shControl cells. Beta-actin was used as loading control.</p

Cox multivariate analysis of factors associated with risk of death.

<p>Cox multivariate analysis of factors associated with risk of death.</p

Activin A immunodetection in OSCC.

<p>(A) Positivity for activin A was observed in the cytoplasm of the tumor cells and in few stromal cells adjacent the tumor. (B) High power view revealed that tumor cells demonstrate variable expression of activin A even in the same tumor. (original magnification: A x100 and B x200).</p

Activin A modulates the adhesion, migration and invasion of OSCC cells.

<p>(A) Activin A treatment induced significantly the adhesion of HaCat cells on surfaces coated with type I collagen, fibronectin or laminin. (B) Follistatin decreased significantly the adhesion of SCC-9 ZsGreen LN-1 cells o surfaces coated with type I collagen and fibronectin. (C) Activin A knockdown augmented the adhesion to coated-surfaces, as revealed by significantly higher adhesion of shINHBA cells compared with shControl cells. (D) Activin A induced significantly the migration of HaCat cells, whereas follistatin blocked it and also reduced significantly the invasion of SCC-9 ZsGreen LN-1 cells through Matrigel-covered surfaces (E). (F) The migration and invasion of shINHBA cells were significantly lower in comparison with shControl cells. (G) Knock down of activin A interferes with cytoskeleton organization, reducing filopodia and lamellipodia formation. The number of filopodia and lamellipodia was significantly lower in shINHBA cells than in shControl cells. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.</p

Activin A controls apoptosis of OSCC cells.

<p>Apoptotic cells were stained with annexin V-PE and propidium iodide and analyzed by flow cytometry. (A) Activin A blocked apoptosis of HaCat cells, reaching significant levels at concentration of 10 and 100 ng/ml. (B) Activin A reduced significantly apoptosis induced by staurosporine. Cells were treated with increased concentrations of activin A for 24 h, but in the last 4 h 0.03 μM of staurosporine were added to the cells. (C) Follistatin significantly induced apoptosis of SCC-9 ZsGreen LN-1 cells. (D) Activin A stable knockdown in SCC-9 ZsGreen LN-1 cells was significantly associated with increased levels of apoptosis. This phenotype was partially rescued adding 100 ng/ml of activin A. Bars represent the means ± SD of three independent experiments. *p>0.05, **p>0.01, ***p>0.005.</p