Significant Role Of Lond Non-coding RNA AFAP1-AS1 In Thyroid Cancer

Part ? Expression of lncRNA AFAP1-AS1 is increased in thyroid cancer tissuesObjective To analyze the expression of lncRNA AFAP1-AS1 in human thyroid cancer tissues and its clinicopathological significance.Methods Real-time quantitative PCR(qRT-PCR)was used to detect the expression of lncRNA AFAP1-AS1 in 36 pairs of thyroid cancer tissues and adjacent normal tissues,and to analyze its clinicopathological significance.Results In 36 cases of thyroid cancer tissues,the expression of AFAP1-AS1 mRNA was significantly higher than that in adjacent nomal thyroid tissues.High expression of AFAP1-AS1 was associated with clinical stages and lymph node metastasis of patients.Conclusion Expression of lncRNA AF AP1-AS1 is increased in thyroid cancer tissues,which means AFAP1-AS1 may act as a oncogene in thyroid cancer.Part ? Down-regulation of lncRNA AFAP1-AS1 inhibits cell proliferation,promotes cell apoptosis and decreases metastasis of thyroid cancer cellsObjective To detect the the expression of AFAP1-AS1 in thyroid cancer cells.To investigate the effects of lncRNA AFAP1-AS1 on cell proliferation,cell apoptosis and cell metastasis of thyroid cancer cells in vitro.To explore the role of AFAP1-AS1 in the development and progression of thyroid cancer preliminarily.Methods The expression of AFAP1-AS1 in thyroid cancer cell lines was detected by qRT-PCR technology.Knocking down expression of AFAP-AS1 in thyroid cancer cell lines FTC 133 and SW579 by AFAP1-AS1 siRNAs,then cell proliferation was determined by MTT and CCK8 assays.The clony formation assay was used to detect cell proliferation ability.The flow cytometry,TUNEL analysis and Western blots assay were used to investigate cell apoptosis of thyroid cancer cells.Transwell technology was performed to evaluate cell invation and migration of thyroid cancer cells.Results The expression of AFAP1-AS1 in thyroid cancer cell lines K-1,TPC-1,SW579,FTC 133 and XTC-1 was significantly higher than that in normal thyroid follicular epithelial cells Nthy-ori3-1.After stably knocking down AFAP1-AS1 in FTC133 and SW579 cells that highly expressed AFAP1-AS1,MTT,CCK8,and the clony formation assays showed that the proliferative ability of thyroid cancer cells decreased significantly.The flow cytometry and TUNEL assays demonstrated increased cell apoptosis after down-expression of AFAP1-AS1.Finally,the Transwell results exhibit decreased invasion and migration ability of thyroid cancer cells in low-expression AFAP1-AS1 cells.Counelusion Expression of AFAP1-AS1 is increased in thyroid cancer cell lines.Down-regulation of AFAP1-AS1 in thyroid cancer cells inhibits cell proliferation,promotes cell apoptosis,and decreases cell metastsis.Part ? LncRNA AFAP1-AS1 regulates the expression of ?-catenin and its nuclear translocation in thyroid cancer cellsObjective To explore the possible mechanisms of AFAP1-AS1 on promoting the invasion and migration of thyroid cancer cells.To clarify the ability of AFAP1-AS1 increases the epithelial-mesenchymal transformation and promotes the invasion and migration of tumor cells by promoting the expression of ?-catenin and its nuclear translocation.Methods Western blot assay was used to detect the transformation of EMT-related markers in thyroid cancer cells after stable knockdown of AFAP1-AS1 in FTC 133 and SW579 cells.The mRNA level of ?-catenin was detected by qRT-PCR.The protein expression of ?-catenin was detected by Western blot assay.Cells were treated with actinomycin,a protein-synthesis inhibitor,then the degradation level of ?-catenin was detected by Western blot.Cells were treated with proteasome inhibitor MG 132,then Western blot and immunoprecipitation assays were performed to detect the degradation pathway of ?-catenin.Finally,.the cell localization of ?-catenin in thyroid cancer cells was detected by immunofluorescence assay.Results After knockdown of AFAP1-AS1 in FTC 133 and SW579 cells,the EMT progression of thyroid cancer cells was inhibited.The expression of E-cadherin,an epithelial marker,was increased while the expression of N-cadherin and Vimentin,two mesenchymal markers,were decreased.The mRNA level of ?-catenin did not have any change while the protein expression level of ?-catenin were significantly decreased.It was found that the degradation rate of ?-catenin was increased after AFAP1-AS1 was knocked down with the treatment of cycloheximide.And ?-catenin was degradated trrough ubiquited-proteasome pathway.The distribution of ?-catenin was translocated from the nucleus to the cytoplasm after AFAP1-AS1 was knocked down.Counclusion Down-regulation of AFAP1-AS1 inhibits the EMT progression of thyroid cancer cells.AFAP1-AS1 had no effects on mRNA level of ?-catenin while reduced its protein expression.AFAP1-AS1 regulateed protein degradation of ?-catenin.AFAP1-AS1 had influence on the distribution of ?-catenin in thyroid cancer cells.