Molecular Mechanisms Of Long Non-coding RNA GHET1 In Regulation Of Invasion And Migration Of Thyroid Cancer Cells

Part Ⅰ The expression of LncRNA GHET1 in tissues of thyroid cancer and its association with the patient clinicopathological characteristics.Objective:The aim of this thesis is to study the expression of LncRNA GHET1 in thyroid cancer tissues and association with patient clinical pathological characteristics.Methods:RT-qPCR was performed to detect the expression of LncRNA GHET1 in papillary thyroid cancer tissues and nontumor tissues from patients.The correlation between the expression LncRNA GHET1 and clinical pathological characteristics of patients with papillary thyroid cancer was analyzed.Result:RT-qPCR assay showed that the expression level of LncRNA GHET1 was increased in papillary thyroid cancer tissues(P<0.05).The increased expression of LncRNA GHET1 was associated with lymphatic metastasis(P<0.05)and TNM stage(P<0.05)of tumor.Conclusion:LncRNA GHET1 expression was significantly increased in papillary thyroid cancer tissues and cells.The high expression of LncRNA GHET1 in papillary thyroid carcinoma tissues was more common in women,and was associated with patients with high TNM stage and positive lymph node metastasis.Part Ⅱ Effect of LncRNA GHET1 on biological characteristics of thyroid papillary cancer cells.Objective:Investigate the effect of LncRNA GHET1 on the invasion,migration,proliferation,and apoptosis of thyroid cancer cells and investigate the underlying mechanism.Methods:1.RT-qPCR was performed to detect the relative expression of LncRNA GHET1 in the papillary thyroid cancer cell lines BCPAP and TPC-1,Nthy-ori3-1 cell line as the control group.2.The effect of knockdown or overexpression of LncRNA GHET1 on invasion,migration,apoptosis and proliferation of thyroid cancer cells was detected by CCK8 assay,clone formation,wound-healing assay,transwell migration and invasion assay and flow cytometry.3.The effect of knockdown or overexpression of LncRNA GHET1 on expression of EMT-related and apoptosis proteins was detected by Western blot.Results:1.The expression of LncRNA GHET1 in TPC-1 and BCPAP cells were higher than those in Nthy-ori3-1 cells(P<0.05).2.CCK8 assay was clone formation assays showed that LncRNA GHET1 promoted the proliferation of papillary thyroid tumor cells(P<0.05).Wound-healing and transwell cell migration assays showed that LncRNA GHET1 promoted the migration ability of papillary thyroid cancer cells(P<0.05).Trans well cell invasion assay showed that LncRNA GHET1 promoted the invasion of papillary thyroid tumor cells(P<0.05).Flow cytometry and cell cycle assay showed that LncRNA GHET1 inhibited the apoptosis(P<0.05).3.Western blot assays showed that apoptosis and EMT related protein E-cadherin in thyroid papillary carcinoma cells increased after knockdown of LncRNA GHET1(P<0.01).The expression levels of Vimentin(P<0.05)and N-cadherin(P<0.01)were decreased.Overexpression of LncRNA GHET1 in thyroid papillary carcinoma cells decreased the expression of apoptosis and EMT related protein E-cadherin(P<0.01),and increased the expression of N-cadherin(P<0.01)and Vimentin(P<0.01).After knockdown LncRNA GHET1 expression,Bax expression increased(P<0.01)and Bcl-2 expression decreased(P<0.01).In contrast,overexpression of LncRNA GHET1 resulted in a decrease in Bax expression and an increase in Bcl-2 expression compared with the control group(P<0.05).Conclusion:LncRNA GHET1 can promote the invasion,migration and proliferation of papillary thyroid carcinoma cells,and inhibit apoptosis.Part Ⅲ The effect of LncRNA GHET1 knockdown on the growth of thyroid cancer cells in nude miceObjective:To investigate the effect of FEZF1-AS1 on the proliferation of colon cancer cells in nude mice.Methods:1.Thyorid papillary cancer cell lines BCPAP was selected to construct a cell line with knockdown LncRNA GHET1.And subcutaneously transplanted tumor model in nude mice was established.2.Compared with the control group,the weight and volume of subcutaneous transplanted tumors from the LncRNA GHET1 knockdown group nude mice were significantly decreased(P<0.05).Result:Compared with the control group,the weight and volume of subcutaneous transplanted tumors from the LncRNA GHET1 knockdown group nude mice were significantly decreased(P<0.05).Conclusion:Knockdown the expression of LncRNA GHET1 inhibited the growth of papillary thyroid cancer transplanted tumors in nude mice.