Expression Of Wnt5a/ROR2 In Thyroid Papillary Carcinoma And Its Effect On Proliferation And Invasion Of Papillary Thyroid Cancer Cells

Part 1 Expression and significance of Wnt5 a in papillary thyroid carcinomaObjective: To investigate the expression of Wnt5 a in papillary thyroid carcinoma and the correlation between Wnt5 a expression and clinicopathological characteristics of patients,and to infer the role of Wnt5 a signaling pathway in the progression of papillary thyroid carcinoma.Methods:1.The expressions of Wnt5 a in papillary thyroid carcinoma tissues and corresponding paracancerous tissues were detected by immunohistochemistry.The correlation between Wnt5 a expression and clinicopathological characteristics of patients was also analyzed.2.The protein expression levels of Wnt5 a in papillary thyroid carcinoma tissues and corresponding paracancerous tissues were detected by western blot.3.The expression levels of Wnt5 a mRNA in papillary thyroid carcinoma tissues and corresponding paracancerous tissues were detected by qPCR.Results:1.The immunohistochemistry assay demonstrated that Wnt5 a was expressed in papillary thyroid carcinoma tissues with the positive rate of 67.2%(39/58),while the positive rate of Wnt5 a expression in corresponding paracancerous tissues was 31.0%(18/58),which was statistically significantly lower than the former(P<0.05).2.The expression level of Wnt5 a in papillary thyroid carcinoma tissues was related to the clinical stage and lymph node metastasis(P<0.05),but not to age,tumor size,sex or multifocality.3.The results of western blot assay demonstrated that the protein expression level of Wnt5 a in papillary thyroid carcinoma tissues(0.98±0.11)was significantly higher than that in corresponding paracancerous tissues(0.42±0.09)(P<0.05).4.The results of qPCR showed that the relative expression of Wnt5 a mRNA in papillary thyroid carcinoma tissues(2.46±0.21)was significantly higher than that in non paracancerous thyroid tissues(1.00±0.12)(P<0.05).Part 2 The Effect of Wnt5 a on Proliferation and Invasion of Papillary Thyroid Cancer CellsObjective: To investigate the effect of Wnt5 a expression on the biological behavior of papillary thyroid carcinoma cells and to analyze its role in the development of papillary thyroid carcinoma.Methods:1.The proliferation of papillary thyroid carcinoma cells with Wnt5 a low-expressed or over-expressed was detected by MTT.2.The clone-forming abilities of papillary thyroid carcinoma cells with Wnt5 a low-expressed or over-expressed were detected by colony formation assays.3.The invasion abilities of papillary thyroid carcinoma cells with Wnt5 a low-expressed or over-expressed were detected by Transwell assays.Results:1.The MTT assays demonstrated that low expression of Wnt5 a could suppress the proliferation of K1 and BCPAP,whereas over-expression of Wnt5 a had promoting effect on proliferation of both kinds of cells(P<0.05).2.The results of colony formation assays showed that the ability of clone formation was obviously enhanced in Wnt5 a over-expressed papillary thyroid carcinoma cells,while low expression of Wnt5 a could repress the clone-forming ability of papillary thyroid carcinoma cells(P<0.05).3.The Transwell assays showed that low expression of Wnt5 a inhibited the invasion abilities of K1 and BCPAP,while over expression of Wnt5 a promoted the invasion abilities of K1 and BCPAP(P<0.05).Part 3 Research on the expression of ROR2 in papillary thyroid carcinoma and its relationship with Wnt5aObjective: To investigate the expression of ROR2 in thyroid papillary carcinoma and the correlation between ROR2 expression and Wnt5 a expression,and to discuss the role of Wnt5 a signaling pathway in the progression of papillary thyroid carcinoma.Methods:1.The expression levels of ROR2 in papillary thyroid carcinoma tissues and corresponding paracancerous tissues were detected by immunohistochemical method and Western blot.The correlation between expressions of ROR2 and Wnt5 a was also analyzed.The association between ROR2 expression and clinicopathological characteristics of patients was analyzed as well.2.The expression levels of ROR2 mRNA in papillary thyroid carcinoma tissues and corresponding paracancerous tissues were detected by qPCR.Results:1.The immunohistochemistry assays demonstrated that ROR2 was expressed in papillary thyroid carcinoma tissues with the positive rate of 62.1%(36/58),while the positive rate of ROR2 expression in corresponding paracancerous tissues was 39.7%(23/58),which was statistically significantly lower than the former(P<0.05).The expression level of ROR2 in papillary thyroid carcinoma tissues was associated with clinical stage and lymph node metastasis(P<0.05),but not with age,tumor size,sex or multifocality.2.Western blot assays demonstrated that the protein expression level of ROR2 in papillary thyroid carcinoma tissues(0.91±0.11)was distinctly higher than that in corresponding paracancerous tissues(0.33±0.06)(P<0.05).3.The results of qPCR showed that the relative expression of ROR2 mRNA in papillary thyroid carcinoma tissues(2.47±0.25)was significantly higher than that in non paracancerous thyroid tissues(1.00±0.14)(P<0.05).4.There was positive correlation between ROR2 expression and Wnt5 a expression(r=0.552,P =0.001).Part 4 Wnt5 a might regulate the invasion and proliferation of papillary thyroid carcinoma cells through ROR2Objective: To detect the effect of ROR2 low-expression combining Wnt5 a over-expression on the proliferation and invasion abilities of papillary thyroid carcinoma cells,to detect the expression of MMP-13 in the Wnt5a/ROR2 signaling pathway.The results could provide experimental basis for the treatment of thyroid papillary carcinoma which targets the Wnt5 a signal pathway.Methods:1.The proliferation ability of papillary thyroid carcinoma cells with ROR2 low-expressed combing Wnt5 a over-expressed was detected by MTT.2.The clone formation ability of papillary thyroid carcinoma cells with ROR2 low-expressed combing Wnt5 a over-expressed was detected by colony formation assay.3.The invasion ability of papillary thyroid carcinoma cells with ROR2 low-expressed combing Wnt5 a over-expressed was detected by Transwell.4.The expression level of MMP-13 in papillary thyroid carcinoma cells with ROR2 low-expressed combing Wnt5 a over-expressed was detected by Western blot.Results:1.MTT assay results showed that low expression of ROR2 inhibited the proliferation of papillary thyroid carcinoma cells.Comparing with Wnt5 a over-expression,the proliferation of papillary thyroid carcinoma cells with Wnt5 a over-expression combing ROR2 low-expression was decreased(P < 0.05).2.The colony formation assay results showed that low expression of ROR2 inhibited the clone formation ability of papillary thyroid carcinoma cells.Comparing with Wnt5 a over-expression,the clone formation ability of papillary thyroid carcinoma cells with Wnt5 a over-expression combing ROR2 low-expression was inhibited(P < 0.05).3.The Transwell assay demonstrated that knocking down of ROR2 suppressed the invasion ability of papillary thyroid carcinoma cells.Comparing with Wnt5 a over-expression,the invasion ability of papillary thyroid carcinoma cells with Wnt5 a over-expression combing ROR2 low-expression was inhibited(P < 0.05).4.The low-expression of ROR2 inhibited the expression level of MMP-13,while over-expression of Wnt5 a increased the expression of MMP-13 in K1 and BCPAP cells.Comparing with Wnt5 a over-expression,the expression level of MMP-13 in papillary thyroid carcinoma cells with Wnt5 a over-expression combing ROR2 low-expression was decreased(P < 0.05).Conclusions:1.The expressions of Wnt5 a and ROR2 in thyroid papillary carcinoma were upregulated and positively correlated.2.Both Wnt5 a ans ROR2 could promote the proliferation and invasion abilities of papillary thyroid carcinoma cells.3.Wnt5 a might regulate the expression of MMP-13 through the ROR2 signaling pathway,thereby promoting the proliferation and invasion of papillary thyroid carcinoma cells.