The Application Of The Double-staining Of E-cadherin And Podoplanin In The Pathology Diagnosis Of The Early-invasive Oral Squamous Cell Carcinoma

Oral cancer is one of the most common malignancies in the world that occurs in human bodies.According to statistics,its incidence is increasing annually.There are about 300,000 new oral cancer patients in the world each year,with a death toll of150,000.And there are about 480,000 new patients and 220,000 deaths each year in China.The WHO statistics show that the incidence of oral cancer ranks sixth in human malignancies,whereas oral squamous cell carcinoma(OSCC)is one of the most common malignancy in oral and maxillofacial malignancies,accounting for approximately 80%of oral and maxillofacial malignancies.In recent years,the incidence showed a trend of rising year by year.OSCC owns strong invasion,which means the surrounding tissues are more easily invaded by the oral squamous cell carcinoma.Despite of improving therapy,the 5-year survival rate remains low at approximately 50%.So the early detection and diagnosis of OSCC is quite important to patients for treatment and prognosis,which would improve survival and quality of life.It is an important process for tumor development from OED to OSCC under the influence of tumorigenic factors.According to the degree of cell atypia,the depth of epithelial layer involved in dysplasia and the infiltration of abnormal cells,we divided them into normal oral epithelial(NOE),oral epithelial dysplasia(OED),squamous Squamous carcinoma in situ(SCIS)and squamous cell carcinoma(OSCC).The methods of treatment to OED and early OSCC were quite different,which means it's important to determine whether the organization is cancer or not in the biopsy tissues.However,the diagnosis of OED and OSCC is quite difficult to identify due to the tissue section and other reasons in the clinicopathological diagnosis,which means that we must explore new ways to help us identify them.Recent studies show that malignant transformation in many carcinomas is associated with the loss of epithelial differentiation and the gain of a mesenchymal phenotype.Epithelial to esenchymal transition(EMT),which had well explain the occurrence,development,invasion and metastasis of tumors.E-cadherin and podoplanin are both important indexes in EMT process.Many studies suggest that E-cadherin may inhibit the invasion of tumor cells,and the reduced expression of E-cadherin is a malignant signal.Lots of research show that the level of the expression of E-cadherin in squamous cell carcinoma(SCC)or at the invasive frontier of the SCC is decreased,which has great significance in the diagnosis of OSCC.Podoplanin can change the actin cytoskeleton of tumor cells or induce collective cell migration by filopodia formation to invade and migrate.Its expression has been found in several tumor cells,including OSCC.The loss of E-cadherin has already been recognized as an important step in the process of tumor progression,while the podoplanin can induce tumorigenesis in the absence of cadherin.The synergy between the two markers is an important approach to the occurrence and progression of tumors.In this study,DIHC was used to detect the expression of E-cadherin and podoplanin in oral squamous epithelium in order to explore the value of the method in the diagnosis of different types of oral squamous epithelial lesions,and judge that whether it is helpful for diagnosis of the early OSCC.ObjectiveBy testing the expression of E-cadherin and podoplanin in oral squamous cell lesion to explore the value of the immunohistochemical double staining in the pathological diagnosis of the early oral squamous cell carcinoma.Methods1.214 cases of Paraffin-embedded tissue specimens were selected in this study,including 30 cases of NOE,66 cases of OED,45 cases of SCIS and 73 cases of OSCC.All the cases were chosen from the biological sample library of the Department of Pathology,the First Affiliated Hospital of Zhengzhou University from2011 to 2015.2.The staining of E-cadherin and podoplanin in all sections was detected by DIHC.3.17 cases tissue specimens whose original diagnoses were SCIS with suspicious infiltration that were examined histologically by hematoxylin and eosin(H&E)staining were selected,then detected the expression of E-cadherin and podoplanin by the DIHC method in the suspicious infiltration area.And telephone follow-up visits were conducted on these cases to track the progress of the disease.4.All statistical analyses were performed using SPSS 21.0 software.Correlations between biomarker co-expression patterns and histopathological features were analysed using the chi-square test.A value of P<0.05 was considered statistically significant.The Bonferroni method was used to correct the level of the inspection When 3 or more groups were analyzed by chi-square test.Results1.In 93.3%of NOE and 62.5%of OED tissues,the expression of E-cadherin showed positive clearly and strongly in cell membrane,while podoplanin was showed negative.The expression of E-cadherin was showed positive in 35.6%of SCIS as the expressions of podoplanin became stronger.The expression of E-cadherin declined obviously and the expression of podoplanin became stronger in the 54.8%of OSCC.With the disease progresses,the expression rate of E-cadherin decreased and the podoplanin positive is increasing.Differences among the four groups were statistically significant(?~2=106.976,P<0.05).2.By the same way,early-invasions were showed clearly in 5 cases(29.4%)of17 indecisive cases.The decrease of E-cadherin and the increase of podoplanin had closely relationship with OSCC.And these 5 cases were verified as OSCC later via re-biopsy or follow-up visits.The coloration of the suspected infiltrate area clearly differs from other tissues may contribute to the rapid identification of the early OSCC.3.The expression of podoplanin of the canceration in OED was strongly positive and the expression of E-cadherin of the canceration decreased or not,comparing with the noncancerous area.4.Significant difference of cytomembrane and cytoplasm coloring of E-cadherin between the OED group and the OSCC group(?~2=13.348,P<0.0167)has been found.The expression of cell membranes is observed in most of the OED,and decreased or negative cell membrane expression in most OSCC.But there was no significant difference in cytoplasmic expression among the three groups.Conclusion1.The cocktail double staining with the combination of E-cadherin and podoplanin in oral squamous epithelial lesion may offer help to the pathological diagnosis and identification of the different squamous epithelial lesions.2.The combination of E-cadherin and podoplanin immunohistochemistry double staining method may improve the diagnostic accuracy and efficiency of the early OSCC or the canceration of OED in focal lesions.3.The DIHC method may help pathologists more quickly locate the suspicious infiltrates and improve diagnostic efficiency.