| Purpose: Thyroid cancer is a common primary and endocrine system tumor in the current clinic.According to the relevant data and information released by the International Health Organization,in recent years,the incidence of this tumor worldwide has been increasing year by year,and the diseased population has gradually shown the characteristics of younger age.Thyroid cancer accounts for 5-10% of women with cancer.Papillary thyroid cancer(PTC)is a subtype of thyroid cancer.According to research,this subtype accounts for more than 90% of thyroid cancer.Clinical studies had shown that patients with this disease have a better prognosis after receiving regular treatment,with a five-year survival rate of over 95%.However,some patients still have a poor prognosis after systemic treatment and radiotherapy and chemotherapy.The symptoms have not improved significantly,and the main manifestations are recurrence and metastasis.According to existing research data,the 10-year survival rate of patients with papillary thyroid cancer is lower than 40%.Compared with other RNAs,long non-coding RNA(lncRNA)has no protein coding function.According to existing studies,the expression of lncRNA is related to the occurrence and development of various diseases in the human body.Long non-coding RNA belongs to the category of antisense RNA.The latest literature reports that AGAP2-AS1 can play a role in promoting tumor occurrence and development in a variety of tumors such as gastric cancer,breast cancer,and non-small cell lung cancer.In this article,the expression mechanism of AGAP2-AS1 in papillary thyroid carcinoma and its role in tumor tissue growth and proliferation are described and studied in detail.At the same time,it further demonstrated the specific mechanism of AGAP2-AS1 regulating cytokines to trigger cancer promotion,and clarified the specific mechanism of its regulation of MMP2.Methods: 1.Collect 110 patients with papillary thyroid carcinoma and paracancerous tissues who were clinically diagnosed with papillary thyroid carcinoma during the period from May 2016 to May 2017 in The First Hospital of China Medical University.Prognosis information and other clinical treatment-related pathological data.2.The expression of AGAP2-AS1 in all sample tissues was counted by q RT-PCR,and the correlation between the expression level of AGAP2-AS1 in papillary thyroid carcinoma tissue and the patients' clinicopathological data was performed based on the chi-square test.Analysis.Transfection of si-AGAP2-AS1 will know the expression level of AGAP2-AS1,and the interference efficiency can be measured based on q RT-PCR.CCK8 can be used to detect the expression level of AGAP2-AS1,and then learn the growth of thyroid papilloma cells,Transwell can explain the expression level of AGAP2-AS1 and its effect on the migration of thyroid papilloma cells,based on the scratch test.The expression of related regulatory factors and the specific impact mechanism of thyroid papillary cell tumor are explained and analyzed.3.Use q RT-PCR and western blot to detect the expression of MMP2 after down-regulation of AGAP2-AS1.4.Detect the expression levels of miRNAs in the NC group and the si-AGAP2-AS1 group based on q RT-PCR,and then screen the miRNAs bound by AGAP2-AS1.miRNA was overexpressed,and western blot was used to detect changes in the expression level of MMP2.5.Use q RT-PCR to detect the expression of miR-424-5p in 40 pairs of papillary thyroid carcinoma tissues and adjacent tissues.Spearman test was used to detect the correlation between the expression levels of AGAP2-AS1 and miR-424-5p in 40 cases of papillary thyroid carcinoma tissues.6.Discuss whether miR-424-5p can bind to the 3'URT of AGAP2-AS1 and MMP2 based on the luciferase reporter gene,and whether AGAP2-AS1 can competitively bind miR-424-5p and then reg?late the expression of MMP2.Results: 1.The expression level of AGAP2-AS1 in papillary thyroid carcinoma cells is higher than that in the adjacent tissues of papillary thyroid carcinoma.It can be seen that the expression of this regulatory factor in papillary thyroid carcinoma is affected by Many factors such as TNM staging are related.2.Inhibition of AGAP2-AS1 in papillary thyroid cancer cells can affect the migration ability of papillary thyroid cancer cells,and effectively change the expression of regulatory factors related to the migration of papillary thyroid cancer cells.3.The expression level of miR-424-5p in papillary thyroid carcinoma tissue is lower than that in adjacent tissues.Overexpression of miR-424-5p in papillary thyroid carcinoma tissue can inhibit the proliferation and metastasis of cancer cells.4.Predict the possible binding of 16 miRNAs to AGAP2-AS1 and MMP2 by using targetscan.Validation based on q RT-PCR found that miRNA-106b-5p,miRNA-20a-5p,miRNA-29b-3p,miRNA-4270,miRNA-424-5p and miRNA-17-5p may bind to AGAP2-AS1.Overexpression of miRNA-106b-5p,i RNA-29b-3p,miRNA-4270,miRNA-424-5p,and miRNA-17-5p in papillary thyroid cancer cells,and MMP2 downstream of miR-424-5p was found to be overexpressed protein expression decreased significantly.5.The luciferase reporter gene showed that miR-424-5p can bind to AGAP2-AS1,MMP2 3'UTR and overexpression of AGAP2-AS1 can bind miR-424-5p to finally realize the regulation of MMP2 expression level.6.Overexpression of miRNA-424-5p can reduce the migration and invasion ability of PTC cells.Simultaneous transfection of si-AGAP2-AS1 and miRNA-424-5p inhibitor can partially rescue the effect of si-AGAP2-AS1 on the migration and invasion ability of PTC cells.inhibition.Conclusion: lncRNA AGAP2-AS1 controls the expression of MMP2 in cells by binding to miR-424-5p specific receptors,which is one of the important factors affecting the metastasis and invasion of papillary thyroid carcinoma. |
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