[Retrospective analysis of clinical and pathological factors that influence patient survival in salivary gland cancer - could chemotherapy improve survival ?].

The purpose of this retrospective study was to evaluate the prognostic factors in salivary gland cancer using a multivariate analysis. In total, 45 consecutive patients who underwent planned radical resection for salivary gland cancer between 1985 and 2010 were analyzed. Univariate and multivariate analyses were performed to analyze the clinical and pathological factors that influence patient survival in salivary gland cancer(tumor location, T and N classification, histological grade, excision margin status, postoperative radiotherapy, and postoperative adjuvant chemotherapy). The results of the multivariate analysis showed that T classification(odds ratio[OR]: 2.93, 95%confidence interval[CI]: 1.47-5.82), and excision margin status(OR: 4.86, 95% CI: 1.76-13.44)had a significant impact on tumor-free survival time, suggesting that both are important prognostic factors in salivary gland cancer. Moreover, postoperative adjuvant chemotherapy showed a tendency to improve tumor free survival time(OR: 0.28, 95% CI: 0.07-1.11), thereby indicating its potential in improving prognosis in patients with salivary gland cancer.

Clinical significance of apoptosis-associated speck-like protein containing a caspase recruitment domain in oral squamous cell carcinoma.

OBJECTIVES: To assess apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) expression in oral squamous cell carcinoma (OSCC) and analyze its clinical and pathological significance. STUDY DESIGN: ASC expression was studied using immunohistochemistry in 119 OSCCs patients. The relationships between ASC expression and clinical and pathological parameters were statistically analyzed. In addition, the relationships between ASC expression and cell differentiation [IVL (involcrin) expression] and apoptosis (TUNEL [TdT-mediated dUTP nick end labeling] positive cell number) were investigated. RESULTS: ASC expression showed significant correlations with parameters including clinical tumor stage, mode of invasion, and histological differentiation, and had a significant impact on survival of OSCC. The distribution of ASC correlated well with that of IVL. ASC expression was significantly correlated with the TUNEL-positive cell number. CONCLUSIONS: Lower ASC expression correlates with clinical and pathological malignancy and, consequently, poor prognosis of OSCC. ASC has a close association with cell differentiation and apoptosis.

Cultured epithelial grafting using human amniotic membrane: the potential for using human amniotic epithelial cells as a cultured oral epithelium sheet.

OBJECTIVE: Human amniotic cells are a valuable source of functional cells that can be used in various fields, including regenerative medicine and tissue engineering. The aim of this study was to investigate the utility of human amniotic epithelial (hAE) cells as a new cell source for culturing stratified epithelium sheets for intraoral grafting. METHODS: Enzymatically isolated hAE cells were submerged in a serum-free, low-calcium-supplemented MCDB 153 medium without a feeder layer. The hAE cells were seeded onto a Millicell cell culture plate insert and cultured while submerged in a high-calcium medium for 4 days. Then, they were cultured at an air-liquid interface for 3 weeks. Cultures of hAE cells proliferated at the air-liquid interface. RESULTS: After 3 weeks, the hAE cells cultivated using the air-liquid interface method lead to almost 10 continuous layers of stratified epithelium without parakeratinization or keratinization. It confirmed immunohistochemically that the presence of CK10/13 and Ki-67 positive cells were spread throughout almost all the epithelial layer, and that CK19 positive cells were expressed throughout the entire epithelial layer in the cultured hAE cell sheets. Cultured hAE cells sheets showed a staining pattern similar to that of uncultured oral mucosa: ZO-1 and occludin were located in the intercellular junctions throughout all the epithelial layers. It was suggested that the hAE sheets consisted of highly-active proliferating cells and undifferentiated cells, and had a barrier function. CONCLUSIONS: These results suggested that hAE cells may be a promising cell source for the development of stratified epithelium allograft sheets using a human cell strain.

Impact of different surgical margin conditions on local recurrence of oral squamous cell carcinoma.

The purpose of this study was to analyze the impact of different surgical margin conditions on local recurrence of oral squamous cell carcinoma (SCC) in 148 consecutive patients who underwent planned radical resection of oral SCC. The patients were classified into four categories according to the status of the surgical margin: clear (no SCC within 5mm, n=103), close (SCC within 5mm, n=21), dysplasia (dysplastic epithelium at margin, n=13), and involved (SCC at margin, n=11). Cox's proportional hazard model showed that the status of the surgical margin had a significant impact on local recurrence (p<0.003); hazard ratio was 3.79 (95%CI: 1.17-12.28) with a close and 7.89 (2.38-26.17) with an involved margin. The presence of mucosal dysplasia at the surgical margin was also a significant predictor of local recurrence (hazard ratio: 5.29, 95%CI: 1.31-21.29). Local recurrence was observed only with severe dysplasia, while no recurrence with mild and moderate dysplasia. In the patients with a clear and closed surgical margin, local recurrence was related with T4 tumor and an advanced mode of tumor invasion. The results of this study suggested that the presence of tumor cells at or close to the surgical margin increased the risk of local recurrence. The presence of dysplastic epithelium (especially severe dysplasia) at the mucosal surgical margin has a significant impact on local control. It was also suggested that not simply the width of the free margin but also clinical and histological risk factors should be included in deciding the necessity for adjuvant radiotherapy.

Intraoperative tissue staining of invaded oral carcinoma.

The purpose of this study was to assess the ability of intraoperative tissue staining with consecutive application of 0.4% indigo carmine and 0.5% Congo red to demonstrate the extent and border of oral carcinoma invasion. Seventeen patients were included in the study. Once the oral tumor was resected, a vertical section of surgical specimen was taken from the central part of the tumor. The extent and border of the invaded carcinoma were assessed on digital microscopic examination with tissue staining. The results of assessments were compared with corresponding results of conventional histopathological analysis with HE staining, which is considered the gold standard. Tissue staining produced a brown-black stain on normal muscle, connective, and salivary tissues but not tumor and epithelial tissues. It clearly demonstrated the extent and border of tumor invasion in 13 of 17 patients (76.5%); however, detection of remnant vital tumor cells in scar tissue after neoadjuvant chemotherapy, and distinction between the tumor and adipose tissue scattered in the muscle tissue was difficult. The results of this study showed that intraoperative tissue staining was a possible method in demonstrating the extent and border of carcinoma deeply invaded in the soft tissue and selecting the site for additional frozen section analysis, although the method needed some refinement.

Accuracy of intraoperative tissue staining in delineating deep surgical margins in oral carcinoma surgery.

The purpose of this study was to assess the accuracy of this tissue staining assessment of surgical specimens in delineating deep surgical margins in oral cancer surgery. Fifteen patients who underwent surgery for oral carcinoma were included in the study. Once the tumor was resected, a vertical section of the surgical specimen was taken from the central part of the tumor. The section was consecutively stained with 0.4% indigo carmine and 0.5% Congo red, and deep surgical margins were assessed using a digital microscope with a magnification power of 25-175x. The results of tissue staining analysis were compared with the corresponding results of conventional histopathological analysis with HE staining, which is considered the gold standard. The extent of carcinoma invasion could be visualized after the application of tissue staining solutions. Tissue staining analysis was accurate in 12 of the 15 patients (80%) in evaluating the closest deep surgical margin. There was no significant difference in the tumor-margin distance between tissue staining and histopathological assessment in these 12 patients (Wilcoxon signed-ranks test, P>0.63). The results of this study showed that intraoperative tissue staining of surgical specimens permitted visual inspection and assessment of tumor spread to surgical margin, although the method has some limitations. The method had a possible ability in controlling the deep surgical margin.

Retrospective analysis on prognostic impact of adjuvant chemotherapy in the patients with advanced and resectable oral squamous cell carcinoma.

PURPOSE: The effect of adjuvant chemotherapy on oral squamous cell carcinoma (SCC) is unclear mainly because there have been a few studies which evaluate the efficacy of adjuvant chemotherapy. The purpose of this retrospective study was to analyze the efficacy of adjuvant chemotherapy in the patients with advanced and resectable oral SCC. METHODS: 41 patients in whom advanced SCC (stage III and IV) was completely removed were included in this study. The impact of multiple variables including T-classification, degree of differentiation, mode of invasion, number and level of cervical metastatic node, pre-and post-operative radiation therapy, neoadjuvant chemotherapy, and adjuvant chemotherapy on survival and control of local relapse or distant metastasis was assessed using the stepwise Cox proportional hazards model. RESULTS: The level of neck node metastasis (p<0.02) was a significant independent predictor for cause-specific survival and adjuvant chemotherapy was of borderline significance (p=0.07). The number of neck node metastasis (p<0.01) and adjuvant chemotherapy (p<0.01) were significantly related with disease free survival. CONCLUSION: The results of this retrospective study suggested that adjuvant chemotherapy had a significant benefit in improving disease free survival.

Intraoperative digital microscopic assessment of the deep surgical margins in oral carcinoma surgery: a preliminary report.

BACKGROUND: This study assessed the possible ability of intraoperative digital microscopic analysis to control the deep surgical margin in oral carcinoma surgery. METHODS: A vertical section of a surgical specimen was taken from the central part of the resected tumor. The section was assessed using a digital microscope. The results of digital microscopic analysis were compared with the corresponding results of the conventional histopathologic assessment. RESULTS: The method was accurate in 10 of the 12 patients (83%) in evaluating the closest deep surgical margin. There was no significant difference in the tumor-margin distance between the digital microscopic and the histopathologic assessments in these 10 patients (Wilcoxon signed-rank test, P > .13). CONCLUSIONS: The digital microscopic examination was useful in controlling deep surgical margins and selecting the nearest margin for additional frozen section analysis. Additional methods that could clarify the difference between the tumor and fibrous tissue are necessary.

Dihydropyrimidine dehydrogenase expression predicts survival outcome and chemosensitivity to 5-fluorouracil in patients with oral squamous cell carcinoma.

Dihydropyrimidine dehydrogenase (DPD) is the rate-limiting enzyme of 5-fluorouracil (5-Fu) catabolism. The aim of this study was to evaluate the prognostic value of DPD expression and the correlation between DPD expression and efficacy of 5-Fu. Retrospective analysis of DPD expression was performed immunohistochemically in 103 patients with oral squamous cell carcinoma (OSCC), in which staining intensity of DPD expression and degree of heterogeneity of DPD expression were categorized. Expression of DPD correlated with lymph node metastasis, mode of invasion and differentiation. Expression of DPD was an independent significant factor for survival outcome and was more predictive than conventional clinical factors. Furthermore, heterogeneous expression of DPD was more effective than homogeneous expression of DPD in neoplastic cells when evaluated in patients treated with chemotherapy including tegafur/uracil (UFT). Expression of DPD is an independent predictor for clinical outcome. Furthermore, heterogeneity of DPD expression may be a clue for predicting sensitivity to 5-Fu in patients with OSCC.

Clinical predictors for contralateral neck lymph node metastasis from unilateral squamous cell carcinoma in the oral cavity.

The factors that predict contralateral (C-) lymph node metastasis (LNM) in patients with unilateral oral squamous cell carcinoma (SCC) were analyzed. One hundred and twenty-nine patients who had untreated SCC that originated from the lateral oral cavity, but not around the midline, were included. The impact of multiple clinicopathologic factors (sex, performance status, primary site, T-stage, number and level of ipsilateral LNM, growth type, histopathological grading, mode of invasion, extension across the midline, and systemic neoadjuvant/adjuvant chemotherapy) on time-to-C-LNM was assessed using the stepwise Cox proportional hazards model. The T-stage, number of ipsilateral LNM, and histopathological grading were independent and significant predictors for C-LNM. No C-LNM occurred in patients without ipsilateral LNM and in those with earlier cancers (T1 to T3) excepting tongue cancer. The results of this retrospective study suggested that patients with advanced tumors, multi-involvement of the ipsilateral neck nodes, or a higher degree of histopathological grading were at a higher risk for C-LNM.

Clinical significance of cellular distribution of moesin in patients with oral squamous cell carcinoma.

PURPOSE: Moesin is a linking protein of the submembraneous cytoskeleton and plays a key role in the control of cell morphology, adhesion, and motility. The aim of the present study was to elucidate the clinical significance of expression patterns of moesin in patients with oral squamous cell carcinoma (OSCC). EXPERIMENTAL DESIGN: Immunohistochemistry for moesin monoclonal antibody was performed on 103 paraffin-embedded specimens from patients with primary OSCC, including 30 patients with locoregional lymph node metastasis, and in the sections from nude mice transplanted with two cell lines derived from a single human tongue cancer (SQUU-A and SQUU-B). RESULTS: Expression patterns of moesin in OSCCs were divided into three groups: membranous pattern; mixed pattern; and cytoplasmic pattern. These expression patterns correlated with tumor size, lymph node metastasis, mode of invasion, differentiation, and lymphocytic infiltration. In about two-thirds of the patients with metastatic lymph node, homogeneous cytoplasmic expression was detected in the metastatic lymph nodes. In addition, SQUU-B with high metastatic potential showed more reduced levels of membrane-bound moesin than SQUU-A with low metastatic potential. A multivariate analysis demonstrated that expression patterns of moesin can be an independent prognostic factor. CONCLUSIONS: Our results suggest that moesin expression contributed to discriminating between patients with the potentiality for locoregional lymph node metastasis and those with a better prognosis and might improve the definition of suitable therapy for each.

Shifts in cellular localization of moesin in normal oral epithelium, oral epithelial dysplasia, verrucous carcinoma and oral squamous cell carcinoma.

BACKGROUND: Moesin, a member of ERM (ezrin/radixin/moesin) family, links actin filaments of cell surface structure to the cell membrane. The purpose of the study is to assess the shifts in cellular distribution of moesin in normal oral epithelium, oral epithelial dysplasia (OED), verrucous carcinoma (VC), and oral squamous cell carcinoma (OSCC). METHODS: The expression of moesin was evaluated immunohistochemically in paraffin-embedded tissues of 59 specimens of OSCC, 35 specimens of OED, 17 specimens of VC, and five specimens of normal oral epithelium. RESULTS: In the normal oral epithelia, all specimens showed a pattern of membranous expression against the anti-moesin antibody in the basal layer cells. In the OED specimens, moesin was dominantly expressed in the cell membrane except for the cornified layer. In VC and OSCC specimens, almost the whole of the carcinoma cells were stained with anti-moesin antibody. However, in OSCC samples, moesin was markedly expressed increasingly in the cytoplasm and decreasingly in the cell membrane, as compared with OED and VC. In addition, there was a significant correlation between the pattern of moesin expression and tumor differentiation in OSCC. CONCLUSIONS: Our results suggest that it is useful to detect the moesin expression as adjunct to screening mucosal lesions in the oral cavity.

[Neoadjuvant chemotherapy with nedaplatin and UFT for oral squamous cell carcinomas].

The purpose of this study was to evaluate the effect of preoperative combination chemotherapy with nedaplatin and UFT for oral squamous cell carcinomas. Eleven patients were enrolled in this study (four men and seven women, with a mean age of 66.3 years). They received oral UFT (300 or 400 mg/day, total 2,100-24,600 mg with an average of 10,700 mg) from the initial diagnosis to the day before surgery. They also received an administration of nedaplatin (80 or 100 mg/m2) about four weeks before the planned surgery. The locoregional response rate (complete response and partial response) was 36.4%, and histological examination of surgical specimens revealed a response rate of 40%. There was no relationship between total dose of UFT and either clinical or histological response. Three patients showed severe neutropenia under 1,000/m3 and all recovered with (two patients) or without (one) the use of G-CSF. No other severe side effect was recognized. From the results of this study it is suggested that this combination therapy is useful for preoperative patients with oral squamous cell carcinoma, and that it is possible for them to undergo the therapy on an out-patient basis.