Overexpression of autophagy-related 16-like 1 in patients with oral squamous cell carcinoma.

Dyregulation of autophagy has been reported in various human cancers including oral squamous cell carcinoma (OSCC). The objective of this study was to link expression of autophagy-related 16-like 1 (ATG16L1), a protein essential for autophagosome formation, to clinical outcome in a cohort of 90 OSCC patients. Expression level of ATG16L1 was assessed by immunohistochemistry and an immunoreactivity score (IRS), ranging from 0 to 9, was assigned to each case. The results were correlated with clinicopathological parameters and outcome of patients. Twenty-seven patients (30%) exhibited ATG16L1 overexpression as indicated by an IRS of 9. Overexpression of ATG16L1 was significantly associated with disease stage (p = 0.001), size (p = 0.031) of the tumor, lymph node metastasis (p = 0.004), and histological grade (p = 0.038). ATG16L1 overexpression significantly affected the overall survival (p = 0.020) and time to recurrence (p = 0.031) of OSCC patients in Kaplan-Meier analysis. The present study suggested that ATG16L1 may be used as a biomarker for selecting OSCC patients with a more aggressive phenotype.

Immunopositivity of Beclin-1 and ATG5 as indicators of survival and disease recurrence in oral squamous cell carcinoma.

AIM: To evaluate the expression and prognostic value of two autophagy-related (Atg) proteins, Beclin-1 and Atg5, in human oral squamous cell carcinoma (OSCC) and to correlate findings with clinical outcomes. PATIENTS AND METHODS: Immunohistochemistry for Beclin-1 and Atg5 was assessed in tumor specimens from 90 patients with OSCC. Immunopositivity was semi-quantitatively scored and receiver-operating characteristic curve analysis was used to determine the cut-off positivity score. RESULTS: 55 (61.1%) and 52 (57.8%) cases showed positive Beclin-1 and Atg5 staining, respectively. 40 tumors (44.4%) were positive for both Beclin-1 and Atg5 expression and 23 cases (25.6%) showed absence of both proteins. Beclin-1 expression significantly correlated with tumor grade (p=0.008) and lymph node metastasis (p=0.009). The expression of Atg5 was associated with tumor grade (p=0.016), advanced clinical stage (p<0.001), large tumor size (p=0.002), and lymph node metastasis (p<0.001). A significant difference in 3-year OS (p=0.050) and TTR (p=0.049) between the patients with Beclin-1 expression and those not showing Beclin-1 expression was found whereas the difference did not reach a statistical significance for Atg5 expression. 3-year OS and TTR differed significantly between patients with dual expression and those with double-negative expression (p=0.022 and p=0.026, respectively). CONCLUSION: Dual expression of tumor Beclin-1 and Atg5 expression may be an adverse prognostic indicator for OSCC.

ATG9A overexpression is associated with disease recurrence and poor survival in patients with oral squamous cell carcinoma.

ATG9A is an integral membrane protein required for autophagosome formation and a membrane carrier in the autophagy pathways. The present study was designed to investigate the expression of ATG9A in oral squamous cell carcinoma (OSCC). Clinically annotated tumor specimens from 90 patients with OSCC were subjected to immunohistochemistry using an antibody against ATG9A and immunoreactivity was scored using an immunoreactivity score (IRS). Scores were compared with clinical and pathologic data to assess association with outcome. Overexpression of ATG9A was defined as an IRS of ≥9 by receiver operating characteristics curve analysis and was identified in 25 (28 %) of 90 cases. ATG9A overexpression was associated with disease recurrence and overall survival (OS) in both univariate (p = 0.030 and 0.025, respectively) and multivariate (p = 0.026 and 0.038, respectively) Cox analyses. Kaplan-Meier plots also showed that patients with ATG9A overexpression had shorter 3-year OS (p = 0.017) and time to recurrence (p = 0.021) than those with low ATG9A expression. These results suggest that the presence of ATG9A in the cytoplasm of tumor cells may be an independent biomarker for disease recurrence and survival in patients with OSCC.

High LC3 expression correlates with poor survival in patients with oral squamous cell carcinoma.

Oral squamous cell carcinoma (OSCC) is a destructive disease with very poor prognosis and no effective treatment. Autophagy is a dynamic cellular process involved in various physiological processes and diseases including cancer that degrades cytoplasmic proteins and organelles. The role of autophagy in the pathogenesis of OSCC is not yet understood. Microtubule-associated protein light chains 3 (LC3) is a reliable autophagosome markers for monitoring autophagy. In the present study, LC3 expression was determined in a cohort of 90 OSCC samples by immunohistochemistry. The results were correlated with clinical and pathological characteristics of patients. High LC3 expression (N = 57; 63.3%) correlated with stage (P < .0001), tumor size (P < .0001), and lymph node involvement (P = .0003) and with an increased risk of death (P < .0001; hazard ratio, 3.59) in a univariate analysis. In the multivariate analysis adjusted for grade, stage, and alcohol, betel, and tobacco consumption, high LC3 expression retained statistical significance with regard to survival (P = .0043; hazard ratio, 2.99). The Kaplan-Meier survival curve also showed that high LC3 expression was significantly associated with poor overall survival (P = .0001). Elevated LC3 expression, which corresponds to increased level of autophagy activity, is a frequent event and an indicator of poor prognosis in human OSCC.

High nuclear protein kinase CßII expression is a marker of disease recurrence in oral squamous cell carcinoma.

BACKGROUND: Protein kinase C beta II (PKCßII) is a member of the family of serine/threonine kinases which are involved in tumor formation and progression. This study investigated the significance of PKCßII in oral squamous cell carcinoma (OSCC). PATIENTS AND METHODS: The expression of PKCßII was determined in tumors from 59 patients with OSCC using immunohistochemistry and was correlated with patients' clinical characteristics and outcomes. RESULTS: Twenty-six cases (44%) exhibited nuclear PKCßII staining. High nuclear PKCßII expression was significantly associated with the consumption of betel quid (p=0.015) and alcohol (p=0.024) in OSCC. Kaplan-Meier analysis revealed a shorter time-to-recurrence in patients with high nuclear PKCßII expression (p=0.018). In multivariate analysis for recurrence, high nuclear staining of PKCßII remained an independent adverse prognostic factor (hazard ratio=2.3, p=0.016). CONCLUSION: The present study provides evidence of the potential prognostic value of PKCßII analysis in OSCC.

High nuclear protein kinase Cθ expression may correlate with disease recurrence and poor survival in oral squamous cell carcinoma.

Protein kinase Cs play important roles in many biological processes and tumorigenesis. This study examined the expression of protein kinase Cθ and assessed its significance in patients with oral squamous cell carcinoma. Immunohistochemical staining was carried out to investigate the expression of protein kinase Cθ in 59 cases of oral squamous cell carcinoma. The results were correlated with clinical characteristics and outcome of patients. Diffuse cytoplasmic protein kinase Cθ was identified in 53 (89.8%) of the 59 oral squamous cell carcinoma cases, and the expression was not statistically associated with any clinicopathologic parameter. Twenty (40.7%) of the 59 oral squamous cell carcinoma cases exhibited nuclear expression of protein kinase Cθ with different grade of intensity. χ(2) analysis indicated that high nuclear protein kinase Cθ expression correlated significantly with shorter 24-month survival (P = .043) and disease recurrence (P = .019). The Kaplan-Meier method also showed that high nuclear expression of protein kinase Cθ was significantly associated with poor overall survival (P = .034) and shorter time to recurrence (P = .003). Univariate analysis revealed that high nuclear protein kinase Cθ expression (P = .046; hazard ratio, 2.2), tumor size less than 2 cm (P = .049; hazard ratio, 4.7), lymph node metastasis (P = .003; hazard ratio, 3.0), and higher stage (P = .002; hazard ratio, 8.7) were each associated with shorter overall survival. We identified the aberrant nuclear expression of protein kinase Cθ in oral squamous cell carcinoma. High nuclear protein kinase Cθ expression may correlate with disease recurrence and poor survival in patients with oral squamous cell carcinoma.

Elevated Krüppel-like factor 4 transcription factor in canine mammary carcinoma.

BACKGROUND: Krüppel-like factors (KLFs) are critical regulators of biological and physiological systems and have been extensively studied for their roles in cell proliferation, differentiation and survival in the context of cancer. Among the KLFs, KLF4 is highly expressed in human breast cancers and plays an oncogenic role. The present study examined the expression of KLF4 and assessed its significance in canine mammary carcinoma. RESULTS: Immunohistochemistry was employed to investigate the expression of KLF4 in 142 cases of canine mammary tumor. 75 of the 142 (52.8%) cases were histologically confirmed as mammary carcinoma. Quantification of immunohistochemistry was carried out using Quick score which multiply the staining intensity by the percentage of positive cells. High KLF4 expression was identified in 44 of the 75 (59%) dogs with mammary carcinoma and none in the benign cases. High KLF4 expression occurred only in the tumor cells and not the adjacent normal cells in mammary carcinoma (P < 0.001). Moreover, the high expression level of KLF4 expression was statistically associated with poor grade, late stage, histological subtypes of simple and complex carcinoma, and shorter 24-month survival. The Kaplan-Meier survival analysis also indicated that dogs with high nuclear KLF4 expression had a significantly shorter survival than those with low/moderate KLF4 expression (P = 0.011). CONCLUSIONS: KLF4 is highly and frequently expressed in canine mammary carcinoma and correlates with a more aggressive phenotype.