Clinical And Experimental Research On The Expression Of MMP-2, CD44v6, P63 In Papillary Thyroid Carcinoma And Their Relationships With Lymph Node Metastases

The relationship between papillary thyroid carcinoma and the expression of protein markers including HBME-1,CK-19, galectin-3 and HER-2/neuObjectives:To investigate the clinical value of differentiation of papillary thyroid carcinoma (PTC) from benign thyroid nodules with human bone marrow endothelial cell (HBME-1), cytokeratin 19(CK-19), galectin-3 and human epidermal growth factor receptor 2 (HER-2/neu).Methods:The retrospective study was done with the parafin sections from the patients including 331 cases with PTC and 664 cases with benign thyroid nodules. All the 4 protein markers were analyzed immunohistochemically.Result:The expression of 4 markers was higher in PTC specimens than that in benign thyroid nodules specimens respectively. The detection of CK-19 had the most sensitivity (92.7%) and that of HBME-1 the most specificity (97.9%) among all of 4 protein markers. The combination of CK-19 and galectin-3 was the most sensitive (82.8%), and the combinations of HBME-1 with CK-19 or galectin-3 or HER-2/neu were the most specific (98.3%). There is no significant in the expression of four markers between papillary thyroid carcinoma with lymph node metastasis and without lymph node metastasis(P>0.05).Conclusions:The combination of HBME-1 and either CK19 or galectin-3 or HER-2/neu can improve the specificity of the diagnosis of PTC. All the four markers might not be useful tumour markers for predicting risk of lymph node metastasis in patients with PTC.Part 2Clinical research on the expression of CD44v6 and MMP-2 and their relationships with lymph node metastasesObjective:To investigate the ability of matrix metalloproteinase-2 (MMP-2) and CD44 variant 6 (CD44v6) protein levels to diagnose papillary thyroid cancer (PTC) and lymph node metastases (LNM) in PTC.Methods:A retrospective study was done with the parafin sections from the patients including 66 cases with PTC,35 cases with benign thyroid nodules and 35 cases adjacent normal follicular epithelium. MMP-2 and CD44v6 protein levels were analysed immunohistochemically using routinely prepared tissue sections.Result:The percentages of samples with cells that demonstrated positive protein staining differed significantly between PTC specimens, benign thyroid nodules and adjacent normal follicular epithelium (MMP-2:86.4%,60.0%, and 25.7%, respectively; CD44v6:80.3%,37.1% and 22.9%, respectively). The expression of MMP-2 and CD44v6 in thyroid papillary carcinoma with lymph node metastasis was significantly higher than without lymph node metastasis.Conclusion:Both MMP-2 and CD44v6 might be useful tumour markers for predicting risk of lateral cervical LNM in patients with PTC.Part 3Experimental research on the expression of CD44+/CD24-~low and p63 and their relationships with the ability of cell migration in PTC cell linesObjective:To explore the relationship between CD44v6 and p63 protein levels and the ability of lymph node metastases (LNM) in PTC cell lines.Methods:Western blot was used to detect the p63 protein expression in a normal thyroid cell line and 4 PTC cell lines including TPC-1, BCPAP, GLAG-66 and NPA. Flow Cytometry was performed to evaluate the proportion of the subpopulation of CD44+/CD24~-low.24-well Transwell test were used to evaluate the ability of cell migration. CCK-8 assays were performed to test the resistance against chemotherapy drug among PTC cell lines.Result:The PTC cell lines with a higher proportion of CD44+/CD24~-low tended to show a high expression of p63, and have significantly strong ability of migration and resistance of chemotherapy drug compared with control team and the lower ones.Conclusion:PTC cells’ ability of migration and invasion may be related to the proportion of CD44+/CD24"-low and expression of p63 in different cell lines. These two markers might be new targets for molecular targeted therapy.