Biological and prognostic significance of NDRG2 downregulation in oral squamous cell carcinoma.

OBJECTIVE: Downregulation of N-myc downstream-regulated gene 2 (NDRG2), a tumor suppressor gene, has been associated with poor clinical outcomes in various cancers. However, the prognostic significance of NDRG2 in oral squamous cell carcinoma (OSCC) remains unknown. This study aimed to evaluate the prognostic value of NDRG2 downregulation in OSCC and to elucidate the mechanism by which NDRG2 is downregulated and the biological role of NDRG2 in tumor progression. METHODS: Immunohistochemical and in silico analyses of NDRG2 expression were performed, and the correlation between NDRG2 expression and clinicopathological data was analyzed. The effect of NDRG2 knockdown on the biological behavior of OSCC cells was investigated and the effect of 5-aza-2'-deoxycytidine (5-aza-dC) on NDRG2 expression was determined. RESULTS: NDRG2 expression was significantly downregulated and DNA hypermethylation of NDRG2 was frequently found in head and neck SCC, including OSCC. Low NDRG2 expression was significantly correlated with adverse clinicopathological features and worse survival in OSCC. NDRG2 knockdown could enhance the oncogenic properties of OSCC cells. NDRG2 mRNA levels in OSCC cells could be restored by 5-aza-dC. CONCLUSION: Downregulation of NDRG2 promotes tumor progression and predicts poor prognosis in OSCC. Therefore, restoration of NDRG2 expression may be a potential therapeutic strategy in OSCC.

Tumor budding is associated with poor prognosis of oral squamous cell carcinoma and histologically represents an epithelial-mesenchymal transition process.

In this study, we aimed to identify whether tumor budding is associated with the progression and prognosis of oral squamous cell carcinoma (OSCC) and investigate the correlation between tumor budding and regulators of epithelial-mesenchymal transition (EMT). Fifty-six cases of OSCC were selected and their tumor budding status was reviewed using archived hematoxylin and eosin-stained slides. In addition, the expression of EMT regulators was evaluated by immunohistochemistry using antibodies against Snail and Twist. Tumor budding was observed in 19 (33.9%) of the 56 cases of OSCC. Tumor budding was strongly associated with lymph node metastasis (P = .001) and shorter overall survival (P = .002). The expression of Snail and Twist was correlated with lymph node metastasis (P < .001 and .002, respectively) and poorer overall survival (P = .024 and .024, respectively). Tumor budding was significantly associated with the expression of Snail (P = .003) and showed a tendency toward higher expression of Twist (P = .08). Therefore, our results suggest that tumor budding is significantly associated with poor prognosis in patients with OSCC and histologically represents an EMT process in OSCC.

Prognostic significance of nestin in primary malignant melanoma of the oral cavity.

Several studies have examined the correlation between nestin expression and the degree of tumor invasion in cutaneous melanoma. However, no information has been reported on nestin in primary mucosal melanoma of the head and neck. The present study examined the expression and prognostic significance of nestin in patients with primary mucosal melanoma of the oral cavity. Nestin expression was examined immunohistochemically in 39 patients (six oral melanoma in-situ cases and 33 invasive oral melanoma cases) and analyzed for association with disease progression. Age, sex, anatomic site, stage, level of invasion, regional lymph node metastasis, surgical margin involvement, and treatment modality were also analyzed. In the 33 invasive melanoma cases, invasion depth correlated significantly with prognosis in univariate and multivariate analyses. High-intensity nestin staining was observed in 14 of the 33 cases and a high proportion of nestin-positive cells was observed in 16 cases. In stage III oral melanoma cases, nestin expression was not significantly associated with disease progression. However, in stage IV cases, both the intensity and the proportion of nestin expression were significantly associated with disease progression (P=0.022 and 0.005, respectively). In all 33 invasive cases, multivariate analyses showed that both the intensity and the proportion of nestin were significantly associated with a poor prognosis (P=0.014 and 0.009; hazard ratio, 3.59 and 4.05; 95% confidence interval, 1.29-9.98 and 1.42-11.56, respectively). In conclusion, nestin can be a valuable prognostic indicator in the advanced-stage (stage IV) cases of oral mucosal melanoma.

KiSS-1 expression in oral squamous cell carcinoma and its prognostic significance.

Downregulated expression of KiSS-1 has been correlated with tumor progression, metastasis, and patient prognosis in various human malignancies. However, there is no information regarding the expression of KiSS-1 in oral squamous cell carcinoma (OSCC). Our aims were to examine KiSS-1 expression in OSCC tissue samples and cell lines and to determine its prognostic significance. KiSS-1 expression was significantly lower in lymph node (LN) metastases than in primary tumor tissues. Five of six OSCC cell lines showed absence or relatively low expression of KiSS-1. Correlations between KiSS-1 expression and clinicopathological parameters were statistically assessed. There were significant correlations between KiSS-1 expression and LN metastasis (p = 0.007), TNM stage (p = 0.024), and local recurrence (p = 0.012). In the Kaplan-Meier survival analysis, negative KiSS-1 expression significantly correlated with poorer overall survival (OS) and disease-free survival (DFS) (p = 0.000 and 0.000, respectively). Multivariate analysis using Cox regression modeling revealed that KiSS-1 expression was an independent prognostic factor for both OS and DFS (p = 0.001 and 0.000, respectively). Our findings suggested that KiSS-1 downregulation may play a role in tumor progression and metastasis of OSCC and may be a reliable biomarker for predicting clinical outcome in OSCC.

Comparative immunohistochemical study of ameloblastoma and ameloblastic carcinoma.

OBJECTIVE: Ameloblastic carcinoma combines the histologic features of ameloblastoma with cytologic atypia, regardless of whether it has metastasized. Because of its rarity, there are few immunoprofile studies of ameloblastic carcinoma and few comparative studies of ameloblastic carcinoma and ameloblastoma. In this study, we compared the expression levels of cytokeratins (CKs), matrix metalloproteinases (MMPs), and Ki-67 between ameloblastoma and ameloblastic carcinoma, and assessed the usefulness of these markers for differentiating the tumors. STUDY DESIGN: We assessed CK7, CK14, CK18, CK19, MMP-2, MMP-9, and Ki-67 expression by immunohistochemistry in 10 cases of ameloblastoma and 7 cases of ameloblastic carcinoma and then compared expression patterns between the 2 groups. RESULTS: Immunostaining for CK14 and CK19 was diffuse and strongly positive in both tumor types, but staining for CK7 was focally positive in only 1 case of ameloblastoma and absent in all cases of ameloblastic carcinoma. However, there was a significant difference in CK18 expression between the 2 tumors (P = .000). Whereas 80% of ameloblastomas showed negative reactivity for CK18, most cases of ameloblastic carcinomas showed a moderate to strong intensity of immunostaining for CK18. Regarding the expression of MMPs, there were significant differences in parenchymal MMP-2 and stromal MMP-9 expression between the 2 tumors. Compared to ameloblastoma, ameloblastic carcinoma showed significantly strong expression of MMP-2 in parenchymal cells (P = .001) and MMP-9 in stromal cells (P = .013). However, there were no differences in MMP-2 expression of stromal cells and MMP-9 expression of parenchymal cells between ameloblastoma and ameloblastic carcinoma. The mean Ki-67 labeling index (LI) of ameloblastic carcinomas was 17.21%, which was significantly higher than that of ameloblastomas (3.57%; P = .002). CONCLUSIONS: The significant expression of CK18, parenchymal MMP-2, stromal MMP-9, and Ki-67 could provide useful markers for differentiating ameloblastic carcinoma from ameloblastoma.

The role of Cripto-1 in the tumorigenesis and progression of oral squamous cell carcinoma.

Oral squamous cell carcinoma (OSCC), the most common malignancy of the oral cavity, remains a lethal disease in over 50% of cases diagnosed annually, due mostly to late detection of this cancer in its advanced stages despite the easy accessibility of the oral cavity for regular examinations. Cripto-1 is a member of the epidermal growth factor (EGF)-CFC protein family and is involved in the activation of several different signaling pathways during embryonic development and cellular transformation. Although the Cripto-1 protein is overexpressed in several human cancers including breast, colon, cervix, gastric, and pancreatic cancer, no prior study has evaluated Cripto-1 expression in OSCC. Therefore, our aims in this study were to examine Cripto-1 expression in clinical samples of OSCC patients using immunohistochemistry, to analyze the correlation between Cripto-1 expression and clinicopathologic parameters, and to identify the oncogenic roles of Cripto-1 in OSCC cell lines. Both epithelial dysplasia (73.3%) and OSCC (55.5%) tissue samples showed significantly higher expression of Cripto-1 than normal mucosa (20%) (p=0.031). In the OSCC samples, there was a significant correlation between Cripto-1 expression and the histological differentiation of OSCC (p=0.015) and a high PCNA index (p=0.011). The in vitro cell proliferation assays demonstrated that recombinant human Cripto-1 (rhCripto-1) induced both SCC-4 and SCC-25 cells to proliferate as compared with control cells (p<0.05 and p<0.01, respectively). In in vitro migration assays, treatment of SCC-4 and SCC-25 cells with rhCripto-1 protein induced a 2.4-fold and 1.7-fold-increase in cell migration, respectively (p=0.000 and p=0.008, respectively). Taken together, our data suggest that Cripto-1 plays a role in the malignant transformation of the oral mucosa and is involved in the tumorigenesis and progression of OSCC by promoting the growth and migration of malignant cells.