Study On The Effect Of S100A12 Gene On Biological Behavior Of Papillary Thyroid Carcinoma And Its Mechanism

objective1.To study S100A12 gene’s differential expression in papillary thyroid carcinoma and normal(peritumorial)tissues;2.To study the relationship between the expression level of S100A12 gene and the gender,age,tumor size,lymph node metastasis,TNM staging of patients with papillary thyroid carcinoma;3.To study the effect of silencing the S100A12 gene on the malignant biological effects(proliferation,invasion and migration)of papillary thyroid carcinoma;4.To preliminarily study the effect of S100A12 gene expression on papillary thyroid carcinoma cells through the changes of EMT pathway related protein expression.Methods1.We collected and screened 120 clinical specimens of patients with papillary thyroid cancer in the First Hospital of China Medical University who had a clinical course of less than 3 years and underwent standard thyroidectomy from 2011 to 2013 to perform immunohistochemical experiments and observe the expression of S100A12 in comparison with peritumorial tissues;2.According to the expression of S100A12,we make statistics and draw charts on relevant clinicopathological characteristics;3.We used Sh RNA to transfect K1 and TPC1 in PTC cell lines to stably silence S100A12 gene,and detected the transfection efficiency by Western blotting.The effect of S100A12 on the proliferation of PTC cells was detected by cell clone formation assay and MTT assay,with transfected noload group cells were used as control.The effect of S100A12 gene silencing on the invasion and migration of PTC cells was evaluated by Transwell invasion and migration assay and cell scratch healing assay;4.Western Blotting was used to detect changes in the expression of related proteins that affect the EMT pathway.Results1.The expression of S100A12 in the cytoplasm of primary papillary thyroid carcinoma and its adjacent tissues was observed after immunohistochemical staining.The results showed that S100A12 was positive in cytoplasm,and the expression level of S100A12 in thyroid cancer tissues was higher than that of the adjacent tissues;2.The investigation showed that high level S100A12 was related to tumor size,TNM grade and lymph node metastasis.However,other clinicopathological features,including age and sex,had no direct relationship with S100A12 expression level;3.The cell clone formation of thyroid papillary carcinoma cell lines K1 and TPC1 after silence of S100A12 showed that the number of clone forming cells in the same time decreased significantly compared with the control group.The results showed that the cell growth curve was inhibited by MTT test after silence of S100A12,indicating that the cell growth was inhibited after silence of S100A12 The proliferation ability of cell lines.The results showed that the invasion and migration of K1 and TPC1 cells were significantly reduced by the number of cells through the membrane penetration,which indicated that the invasion and migration of the cells decreased.Compared with the control group,the migration rate of thyroid papillary carcinoma cell lines K1 and TPC1 was decreased significantly,which indicated that the cell migration ability was decreased;4.S100A12 may enhance the invasion and migration of thyroid papillary carcinoma cells by inducing EMT.The expression of E-cadherin and N-cadherin in EMT signal pathway was detected by Western blotting.After Sh S100A12 was transfected into TPC1 and K1 cells,the expression level of E-cadherin increased,while the expression level of N-cadherin decreased.Conclusion1.S100A12 is up-regulated in papillary thyroid cancer.The expression of S100A12 was related to tumor size,TNM grade and lymph node metastasis;3.The silent S100A12 gene inhibits the proliferation,invasion and migration of thyroid cancer cells;4.S100A12 may promote the invasion and migration of thyroid papillary carcinoma cells by inducing EMT.