The relationship of the histologic grade at the deep invasive front and the expression of Ki-67 antigen and p53 protein in oral squamous cell carcinoma

BACKGROUND: Although many histopathologic characteristics of oral squamous cell carcinoma (O-SCC) have been identified as prognostic factors, accurate, and unequivocal factors have not been clearly identified. The purpose of this study was to evaluate a potential association between the histologic grade of malignancy at the deep invasive front and the expression of Ki-67 antigen and p53 protein in O-SCC. METHODS: The expression of Ki-67 antigen and p53 at the invasive tumor front area of O-SCC was examined by immunohistochemistry of archived tissue from 62 cases. The mean age of patients was 60.7 years (range: 37-89) and the male-female ratio was 1.6:1 (38 men, 24 women). There were 20, 17, 14, and 11 cases classified as stage I to stage IV, respectively. The correlation between the intensity of immunostaining for Ki-67 antigen and p53 and the histologic grade of malignancy at the deep invasive front (invasive front grade, IFG) was analyzed. The expression of Ki-67 antigen and p53 in normal oral epithelia (10 cases) was also investigated. RESULTS: The mean Ki-67 labeling index (LI) in the O-SCC samples was 32.8 +/- 12.0% (n = 62). The mean total score of IFG (IFG score) was 9.1 +/- 2.7 points (n = 62). There was a significant linear correlation between the IFG score and the Ki-67 antigen (gamma = 0.651, R2 = 0.596, P < 0.0001). Of 50 tumors examined, 27 (54.0%) exhibited p53-positive nuclear immunostaining. The staining patterns for Ki-67 antigen and p53 were similar. Both Ki-67-LI and p53-positive status were significantly correlated with the IFG scores. CONCLUSION: The findings of this study demonstrate that overexpression of Ki-67 antigen and p53 at the deep tumor invasive front of O-SCC is associated with histologic grade of malignancy.     

Reduced syndecan-1 expression is correlated with the histological grade of malignancy at the deep invasive front in oral squamous cell carcinoma

BACKGROUND: Although many histopathological characteristics of oral squamous cell carcinoma (O-SCC) have been identified as prognostic factors, no factor is completely accurate and unequivocal. This study evaluated the association between the loss of syndecan-1 expression and the histological grade of malignancy at the deep invasive front in O-SCC. METHODS: The expression of syndecan-1 at the invasive tumor front of O-SCC was examined immunohistochemically using archived tissue from 72 cases. The mean age of the patients was 62.5 years (range: 23-90 years) and the male-female ratio was 1.3:1 (41 men, 31 women). There were 26, 24, 11, and 11 cases classified as stages I-IV respectively. The correlation between the intensity of syndecan-1 immunostaining and the clinicopathological factors, especially the histological grade of malignancy at the deep invasive front (invasive front grade) was analyzed. RESULTS: Of the 72 cases, seven (9.7%), 29 (40.3%), 36 (50.0%) showed strong, intermediate, and weak or negative syndecan-1 staining respectively. There were significant differences between syndecan-1 expression and prognosis, differentiation, and pattern of invasion at the deep invasive front. Moreover, the invasive front grade scores, based on the intensity of syndecan-1 staining, were 5.6 +/- 1.0, 8.0 +/- 2.1, and 10.2 +/- 2.3 points with strong, intermediate, and weak or negative intensity respectively; and the difference was significant (P < 0.0001). Patients with intermediate or strong intensity for syndecan-1 had significantly better prognoses than did those with negative or weak intensity (P = 0.0138). CONCLUSION: This study demonstrated that the reduced expression of syndecan-1 seems to be a useful marker of histological malignancy at the deep tumor invasive front and may be a useful prognostic factor in O-SCC.     

Immunohistochemical expression of secreted frizzled receptor protein 1 in the invasive front of tongue squamous cell carcinoma

The Wnt signalling pathway is involved in the development of oral cancer. When inactivated, secreted frizzled receptor protein 1 (SFRP1), a cellular Wnt‐inhibitor protein, may enhance cancer development. The aim of this study was to assess the expression of SFRP1 in the invasive front of tongue squamous cell carcinoma (SCC). Immunohistochemical expression of SFRP1 in the cohesive and discohesive invasive front of 36 resection specimens of tongue SCC was investigated. Immunostaining was compared with tumour size, tumour thickness, and neural invasion. Immunoreactivity increased in depth from the surface of the tumour to the invasive front. There was a statistically significant association between type of invasive front and pattern of SFRP1 expression. A discohesive invasive front showed more intense immunoreactivity. No statistically significant associations were found between the intensity of staining and relevant prognostic factors. Tongue SSC has variable SFRP1 expression in different parts of the tumour and the greatest expression is in the invasive front. The more intense staining in the discohesive invasive front compared with the cohesive invasive front might contribute to the worse prognosis of cancers with a discohesive invasive front.     

Analysis of the Ki-67 antigen at the invasive tumour front of human oral squamous cell carcinoma

BACKGROUND: It is hypothesised that cell proliferation, as measured by the Ki-67 labelling index (LI) at the invasive tumour front (ITF) was directly related to the histological grade in human oral squamous cell carcinomas (SCCs). METHODS: Tissues from 42 human oral SCCs were collected and stained with an antibody directed against the Ki-67 antigen using an advanced polymer staining system. Quantitation of the immunopositive cells was performed on two parallel sections at the invasive tumour front (ITF), using an image analyser. The Ki-67 LI was expressed as the number of positive nuclei/mm2 of epithelium. The control tissue used was normal epithelium at the excision margin. RESULTS: The mean Ki-67 LI for oral SCCs at the ITF was significantly greater than that for the excision margin tissue (P < 0.0001). There was a positive association between increasing Ki-67 LI and increasing Broders' grade (P < 0.05), with a well-differentiated tumour having the lowest mean Ki-67 LI (1549 +/- 806) and a poorly differentiated tumour having the highest value (2232 +/- 771). A similar trend was observed between the mean Ki-67 LI and Bryne's multifactorial grading system. CONCLUSIONS: It was concluded from this study that cell proliferation (as measured by the Ki-67 antigen) at the ITF had a strong positive relationship with histological grading in human oral SCC.     

Tumor budding correlates with poor prognosis and epithelial‐mesenchymal transition in tongue squamous cell carcinoma

J Oral Pathol Med (2011) 40 : 545–551 Background: Tumor budding is a readily detectable histopathological feature and has been recognized as an adverse prognostic factor in several human cancers. However, the prognostic value of tumor budding in tongue squamous cell carcinoma (TSCC) has not been reported. The purpose of this study was to assess the correlation of tumor budding with the clinicopathologic features, and the known molecular biomarkers (E‐cadherin and Vimentin), as well as to evaluate its prognostic significance for TSCC. Methods: Archival clinical samples of 230 patients with TSCC were examined for tumor budding. Immunohistochemistry analyses were performed to examine the expression of E‐cadherin and Vimentin. Statistical analyses were carried out to assess the correlation of tumor budding with clinicopathologic parameters and patient survival. The potential association between tumor budding and alterations of E‐cadherin and Vimentin expression was also assessed. Results: Of the 230 TSCC cases examined, tumor budding was observed in 165 cases (71.7%), with a mean tumor bud count of 7.5 (range from 1 to 48 buds). High‐intensity budding (≥5 tumor buds) was observed in 111 cases (48.3%). Statistical analysis revealed that tumor budding was associated with tumor size (P < 0.05), differentiation (P < 0.05), clinical stage (P < 0.05), lymph node metastasis (P < 0.01), and correlated with reduced overall survival. In addition, significant associations were observed among tumor budding and the deregulation of E‐cadherin (P < 0.001) and Vimentin (P < 0.001). Conclusions: Tumor budding, which associates with epithelial–mesenchymal transition, is a frequent event and appears to be an independent prognostic factor in TSCC.     

Expression form of p53 and PCNA at the invasive front in oral squamous cell carcinoma: correlation with clinicopathological features and prognosis

J Oral Pathol Med (2011) 40 : 693–698 Background: Abnormalities in cell‐cycle‐controlling genes are important in the malignant transformation and proliferation of tumors. Among these genes, the tumor suppressor gene p53 is the most notable, and its mutations provide an indicator of tumor progression and prognosis. Proliferating cell nuclear antigen (PCNA) is a highly conserved nuclear protein that is expressed during cell replication and DNA repair. This study examined the expression of p53 and PCNA at the invasive front of oral squamous cell carcinomas (OSCC) by immunohistochemical staining, and investigated the relationship of these proteins to clinicopathological findings and prognosis. Methods: Fifty‐nine biopsy cases of OSCC were examined by immunohistochemical staining. Clinicopathological data were gathered and patient survival was analyzed. Results: The p53 labeling index (p53‐LI) and PCNA labeling index (PCNA‐LI) were examined at the invasive front of the tumors. A high p53‐LI (p53+) was observed in 17 of the 59 cases (28.8%) and a high PCNA‐LI (PCNA+) was observed in 28 of the 59 cases (47.5%). Among the modes of cancer invasion, many of the p53+/PCNA+ cases could be confirmed as highly invasive cancer (P < 0.05). In addition, the p53+/PCNA+ cases showed a high risk of tumor recurrence compared with the other expression forms, and patients with p53+/PCNA+ had a worse prognosis than those with the other expression forms. High labeling indices of p53 and PCNA are associated with poor prognosis in patients with OSCC. Conclusion: We suggest that it is important to investigate the expression of p53 and PCNA at the invasive front of OSCC.     

舌鳞状细胞癌浸润前沿细胞增殖的研究

目的　研究舌磷癌浸润前沿分级、检测增殖细胞核抗原 (PCNA)、Ki6 7、核仁组织区相关嗜银蛋白 (AgNOR)在浸润前沿中的表达。 方法　对 5 7例舌鳞癌行浸润前沿分级 ,采用免疫组化SP法及银染色法检测浸润前沿和中心PCNA、Ki6 7、NORs的表达。结果　浸润前沿PCNA、Ki6 7的表达和AgNOR均数均明显高于非前沿部分 ,差异有极显著性 (P <0 .0 0 1) ;浸润前沿PCNA标记指数 (P <0 .0 5 )、AgNOR均数 (P <0 .0 1)与浸润前沿分级 (IFG)总分呈正相关关系 ;单因素COX分析显示 ,IFG总分 (P <0 .0 1)、浸润前沿AgNOR均数 (P <0 .0 5 )对判断预后有意义 ;多因素COX分析显示 ,IFG总分 (P <0 .0 1)可作为预测总体生存率和累积生存率的有意义的因子。结论　舌鳞癌浸润前沿肿瘤细胞分化较差 ,增殖活性明显高于非前沿部分 ;IFG总分和浸润前沿AgNOR均数与舌癌预后有关     

The importance of stromal inflammation in squamous cell carcinoma of the tongue

J Oral Pathol Med (2012) 41 : 379–383 Background: Histological risk assessment evaluating worst pattern of tumour invasion (WPOI), and lymphocytic response (LR), has previously been shown to be of prognostic significance in squamous cell carcinomas of the head and neck (SCCHN). SCCHN is a heterogeneous group of tumours including tumours located in the oral cavity, of which the majority is located in the tongue. Methods: Haematoxylin/eosin–stained slides from diagnostic biopsies from 94 cases of SCC on the tongue were evaluated for WPOI and LR. Within the inflammatory infiltrate, the percentage of eosinophilic granulocytes was also estimated. Results were correlated with clinical data such as response to treatment and recurrence. Results: For WPOI the majority of patients, 84%, showed small invasive tumours islands with a size <15 cells (grade 4). No correlation with survival, response to treatment or recurrence was seen for WPOI. More than half of the patients showed a dense lymphocytic infiltrate, a factor that was significantly correlated with complete response to radio therapy. Of the patients with dense lymphoid infiltrate, the majority, 63%, did not either have a recurrence. No significant correlation with recurrence, response to treatment or any other factor was seen for presence of eosinophils. Conclusions: Data clearly showed that tongue tumours have a split invasive growth pattern and an intense inflammatory response at the tumour interface. Results also indicated that evaluation of the intensity of the inflammatory infiltrate at the tumour interface in tongue SCC could provide information of potential importance for choice of treatment and prognosis.     

Evaluation of survivin as a prognostic marker in oral squamous cell carcinoma

J Oral Pathol Med (2010) 39 : 368–375 Background: Poor prognosis of oral squamous cell carcinoma (OSCC) is partly attributed to the lack of significant tumor marker for accurate staging and prognostication. We have evaluated survivin, which is a member of the inhibitor of apoptosis family as a cancer marker associated with proliferation, angiogenesis, oral carcinogenesis, and OSCC patient survival, as we reported a prognostic significance of survivin expression in lymph node previously. Methods: To evaluate survivin expression in six OSCC cell lines, Western blotting was performed. Hamster oral carcinogenesis model was used to observe changes of survivin expression in oral carcinogenesis. Finally, we assessed the diagnostic and prognostic significance of survivin in a series of 38 primary OSCC through immunohistochemistry (CD31, PCNA) and Kaplan–Meier’s test. Results: Survivin expression was detected in all OSCC cell lines at a varying level but not observed in normal gingival keratinocyte cells. In hamster model, survivin expression was observed from 8 weeks through 16 weeks and the intensity of expression became strong until 16 weeks. Clinicopathological analysis revealed a significant correlation between survivin expression and lymph node metastasis (P = 0.006) and proliferation (P < 0.001). However, there was no significant relationship with differentiation, micro vessel density, and cancer stage based on TNM. Survivin overexpression had a significant negative effect on survival of patients. Conclusions: These results demonstrate the significant relationship between survivin expression and oral carcinogenesis and aggressiveness of OSCC including survival rate of patient. Survivin therefore may be used as a significant cancer marker to gain prognostic information of OSCC.     

磁共振成像评估舌鳞状细胞癌浸润深度的准确性分析

目的以光镜下病理切片为参照,分析磁共振成像(magnetic resonance imaging,MRI)测量的舌鳞状细胞癌浸润深度的准确性,为临床提供参考。方法选取2018年1月至2020年9月就诊于山西医科大学第一医院口腔科和中南大学湘雅口腔医院的73例舌鳞状细胞癌患者,术前均行MRI评估舌鳞状细胞癌浸润深度,术中冰冻病理切片再次测量舌鳞状细胞癌浸润深度。结果T1加权成像测量的舌鳞状细胞癌浸润深度较病理结果平均高估1.11 mm(95%CI=0.51~1.70,t=3.72,P<0.001),相关系数r为0.95;T2加权像平均高估2.17 mm(95%CI=1.32~3.02,t=5.10,P<0.001),相关系数r为0.92。Bland?Altman图显示T1、T2加权像与病理测量的浸润深度一致性佳。结论MRI测量的舌鳞状细胞癌浸润深度较为准确,与病理测量结果相比有平均1~2 mm的高估,其中T1加权像优于T2加权像。     

The relationship of proliferating cell density at the invasive tumour front with prognostic and risk factors in human oral squamous cell carcinoma

BACKGROUND: We hypothesise that the density of proliferating cells at the invasive tumour front (ITF) has a positive relationship with prognostic and risk factors in human oral squamous cell carcinoma (SCC). METHODS: Tissues from 47 human oral SCC specimens were collected and stained with a monoclonal antibody directed against the Ki-67 antigen using a horseradish peroxidase based two-step immunostaining method. Counting was performed on two parallel sections at the ITF using an image analyser. The Ki-67 labelling index (LI) was determined by measuring the number of nuclei/mm(2) of epithelium. RESULTS: Our results show that the density of proliferating cells is related to clinical staging, with advanced stage of disease having a significantly higher Ki-67 LI compared with early stage of disease (2111 +/- 905 vs. 1908 +/- 913; P = 0.03). Importantly, this study shows that tumours that have metastasised have a significantly higher Ki-67 LI than tumours where distant metastasis was not detected (3257 +/- 650 vs. 1966 +/- 881; P < 0.0001). CONCLUSIONS: Cell proliferation, as measured by the Ki-67 LI at the ITF, has a positive relationship with clinical staging, tumour thickness, smoking status of the patient and alcohol consumption. Further, we suggest that a multicenter study with a large cohort of patients is indicated to fully elucidate whether cell proliferation at the ITF is directly related to patient survival.     

实时荧光定量RT-PCR检测舌癌E-cadherin mRNA的表达

目的:了解E-cadherin在舌癌中的表达及其临床意义.方法:采用实时定量PCR检测29例舌鳞癌患者的癌组织和正常组织中E-cadherin mRNA,分析E-cadherin基因表达与临床病理参数的相关性.结果:舌癌组织中E-cadherin mRNA表达水平2.23±1.16(E-cadherin/β-actin),低于正常组织组8.59±2.71,两组间差异有统计学意义(P＜0.01),E-cadherin mRNA水平与临床病理参数之间无相关性(P＞0.05).结论:E-cadherin的表达下降是舌癌发生过程中的重要事件,实时定量PCR检测E-cadherinmRNA的表达对舌癌早期诊断有重要参考价值.     

EMMPRIN expression in tongue squamous cell carcinoma

Background:  Extracellular matrix metalloproteinase inducer (EMMPRIN) is identified as a tumor-cell membrane protein that stimulates matrix metalloproteinases (MMPs) production. Several studies have shown that higher EMMPRIN expression is associated with shorter survival time and correlated significantly with more advanced clinico-parameters of cancer. The aim of this study was to investigate the relationship between clinico-pathologic characteristics and EMMPRIN, and prognostic significance of EMMPRIN expression in human tongue squamous cell carcinoma.
Methods:  Extracellular MMP inducer expression was examined immunohistochemically on paraffin-embedded tissue specimens from 68 patients with tongue squamous cell carcinoma and who underwent radical surgeries from 1996 to 2006. The 68 patients were followed up from 1 to 119 months, with an average of 27.5 months. Nonparametric tests were performed for the comparison of EMMPRIN expression between two independent groups. Survival analysis was performed to find the prognostic significance of EMMPRIN expression.
Results:  We found that EMMPRIN expression in tongue squamous cell carcinoma is significantly higher than that in non-cancerous epithelium adjacent to carcinoma of tongue. In addition, EMMPRIN expression is significantly associated with tumor diameter and clinical stage in the samples, but did not correlate with gender, age, tumor metastasis, and pathological grade. Finally, survival analysis indicates that EMMPRIN overexpression correlates significantly with poor overall survival in the patient cohort.
Conclusion:  These results suggest that EMMPRIN might represent an attractive target for immunotherapeutic approaches in a subgroup of patients with tongue squamous cell carcinoma.