Abdalla Zahra, Walsh Tanya, Thakker Nalin, Ward Christopher M
Stem Cell Research Group, Manchester Dental School, Manchester, United Kingdom.
Department of Histopathology, Manchester Royal Infirmary, Manchester, United Kingdom.
PLoS One. 2017 Dec 7;12(12):e0187449. doi: 10.1371/journal.pone.0187449. eCollection 2017.
Oral squamous cell carcinoma (OSCC) is a highly aggressive cancer that is associated with poor 5-year patient survival. Disease treatment is further compounded by the difficulty in predicting pre-cancerous tissues that will progress to OSCC and the high recurrence rates following surgical resection. Here we have assessed expression of the oral epithelial markers E-cadherin, EMP1 and 5T4 and the pro-invasive N-cadherin proteins using fully characterised antibodies and quantitative immunofluorescence microscopy in normal tissue (NT), fibroepithelial polyp (FEP), low-grade dysplasia (LGD), high-grade dysplasia (HGD), T1 OSCC and T4 OSCC biopsies. Decreased E-cadherin expression was associated with FEP, LGD and HGD biopsies, demonstrating that loss of E-cadherin is an early event within abnormal epithelium and occurs in the absence of an E- to N-cadherin switch, the latter of which was only observed in T4 OSCC. Furthermore, loss of E-cadherin and EMP1 is an indicator of LGD (p = 0.0006) and loss of E-cadherin, EMP1 and 5T4 an indicator of HGD (p = 0.0006). Expression patterns of E-cadherin, EMP1 and N-cadherin could predict abnormal epithelium in LGD, HGD, T1 and T4 OSCC biopsies (z-value = 0 for all disease grades) and allowed classification of LGD (z = 1.47), HGD (z = 2.138), T1 (z = 1.05) and T4 OSCC (z = 1.49) biopsies. Therefore, these markers provide a useful means to predict abnormal epithelium in patient biopsies. Linear regression and coefficient of determination analysis revealed positive correlation with a NT>LGD>HGD disease transition but low correlation with a putative HGD>T1 OSCC>T4 OSCC disease transition. Furthermore, expression of E-cadherin, EMP1, 5T4 and N-cadherin in pathologically normal surgical safety margins of LGD, HGD and T1 OSCC patient biopsies revealed significant differences to NT and the use of safety margins or FEP as 'normal tissue' controls introduced Type II errors in all patient cohorts. This work forms the basis for further investigation of the role of E-cadherin loss in abnormal epithelium and in the development of automated analyses for use in cancer diagnostics.
口腔鳞状细胞癌（OSCC）是一种侵袭性很强的癌症，患者5年生存率较低。由于难以预测哪些癌前组织会发展为OSCC，以及手术切除后的高复发率，疾病治疗变得更加复杂。在这里，我们使用经过充分表征的抗体和定量免疫荧光显微镜，评估了正常组织（NT）、纤维上皮息肉（FEP）、低级别发育异常（LGD）、高级别发育异常（HGD）、T1期OSCC和T4期OSCC活检组织中口腔上皮标志物E-钙黏蛋白、EMP1和5T4以及促侵袭性N-钙黏蛋白的表达。E-钙黏蛋白表达降低与FEP、LGD和HGD活检组织相关，表明E-钙黏蛋白的缺失是异常上皮内的早期事件，且在E-钙黏蛋白向N-钙黏蛋白转换缺失的情况下发生，后者仅在T4期OSCC中观察到。此外，E-钙黏蛋白和EMP1的缺失是LGD的一个指标（p = 0.0006），E-钙黏蛋白、EMP1和5T4的缺失是HGD的一个指标（p = 0.0006）。E-钙黏蛋白、EMP1和N-钙黏蛋白的表达模式可以预测LGD、HGD、T1期和T4期OSCC活检组织中的异常上皮（所有疾病分级的z值均为0），并能够对LGD（z = 1.47）、HGD（z = 2.138）、T1期（z = 1.05）和T4期OSCC（z = 1.49）活检组织进行分类。因此，这些标志物为预测患者活检组织中的异常上皮提供了一种有用的方法。线性回归和决定系数分析显示，与NT>LGD>HGD的疾病转变呈正相关，但与假定的HGD>T1期OSCC>T4期OSCC的疾病转变相关性较低。此外，在LGD、HGD和T1期OSCC患者活检组织的病理正常手术切缘中，E-钙黏蛋白、EMP1、5T4和N-钙黏蛋白的表达与NT存在显著差异，并且使用手术切缘或FEP作为“正常组织”对照在所有患者队列中都引入了II类错误。这项工作为进一步研究E-钙黏蛋白缺失在异常上皮中的作用以及开发用于癌症诊断的自动化分析奠定了基础。
