| Objective: Thyroid cancer is the most common endocrine tumor,accounting for about 1% of all malignant tumors.The pathological types of thyroid cancer include papillary carcinoma,follicular carcinoma,undifferentiated carcinoma and medullary carcinoma.Among them,papillary carcinoma is the most common,with low malignancy and good prognosis.The incidence of thyroid cancer is related to age,gender and region.In recent years,the incidence of thyroid cancer has been on the rise,and the neck lymph node metastasis has a strong trend.Treatment options for patients with advanced or recurrent metastatic disease are limited,leading to poor prognosis.Therefore,studying the molecular mechanism of the occurrence and development of papillary thyroid carcinoma is of great significance for finding new treatment options and targets.TRIM29 and FN1: TRIM29 is maladjusted in a variety of tumors and plays a role in promoting or inhibiting cancer.In this study,we found increased expression of TRIM29 and fibronectin 1(FN1)in PTC tissues.Overexpression or downregulation of TRIM29 did not significantly alter the proliferation of PTC cells.TRIM29 overexpression significantly promoted,while TRIM29 knockdown significantly reduced migration and invasion by regulating FN1 expression in PTC cells.We found that TRIM29 altered the stability of FN1 mRNA by regulating the expression of mi R-873-5p.This study also demonstrated that longchain non-coding RNA(Lnc RNA)CYTOR inhibits the maturation of mi R-873-5p by interacting with premi R-873,and TRIM29 inhibits mi R-873-5p by upregulating CYTOR.This study helps us to further understand the potential mechanism of PTC transfer,and suggests TRIM29 may be a new therapeutic target and a potential intervention strategy for the PTC.Methods: 1.Use Western blot experiment to analyze the expression of TRIM29 in three thyroid cancer cell lines,construct a stable TRIM29 cell line,and then perform cell counting and Transwell experiments to verify the effects of TRIM29 on the proliferation and invasion and migration capabilities of thyroid cancer cells.2.Use Western blot experiment to analyze the relationship between FN1 and TRIM29 in thyroid cancer tissues and cells,and construct FN1 overexpression or knockdown after stabilizing the cell line,verify whether the cell’s proliferation,invasion and migration capabilities have changed.3.RT-PCR was used to analyze the effect of TRIM29 on FN1 mRNA synthesis,Western blot and RIP experiments were used to analyze the effect of TRIM29 on AGO2 protein expression and the enrichment of FN1 mRNA.4.Through the online database(starbase.sysu.edu.cn),analyze the coexpression of TRIM29 and has-mi R-873-5p in papillary carcinoma tissues,using mi R-873-5p antagomir and mi R-873-5p mimic treated the IHH4 and TPC1 cell lines stably transformed with TRIM29,and used Western blot to verify whether FN1 was affected and its effect on cell invasion and migration.5.Using real-time RT-PCR to analyze the expression levels of premi RNA-873,primi R-873 and mi R-873-3p in the TRIM29 stable cell line,and verify the expression of TRBP and Dicer in the stable TRIM29 cell line.6.According to the online database(starbase.sysu.edu.cn),analyze the correlation between CYTOR and TRIM29 and FN1,and the correlation between TRIM29 mRNA and lnc RNA CYTOR.7.The cell distribution of TRIM29 and lnc RNA CYTOR in IHH4 cells were studied by immunofluorescence and FISH staining.8.After knocking down or overexpressing CYTOR in thyroid cancer cells,the changes in FN1 protein expression were verified by Western blot,and its effect on invasion and migration was further verified.9.Show the potential binding site between premi R-873 and CYTOR according to Lnc Tar,and confirm their interaction.10.Analyze the effect of knocking down CYTOR on the expression of premi R-873,mi R-873-5p and mi R-873-3p in TRIM29 cells by q RT-PCR analysis.11.According to the online database(starbase.sysu.edu.cn),analyze the co-expression of CYTOR,hsa-mi R-873-5P,hsa-mi R-873-3P in papillary carcinoma tissues.Results:1.TRIM29 overexpression or knockdown promoted or suppressed migration and invasion of PTC cells,respectively.2.TRIM29 was positively corrected with FN1,and regulated the invasive capacities of PTC cells by regulating FN1.3.TRIM29 did not affect de novo synthesis of FN1 mRNA,but altered FN1 mRNA stability through Ago2-dependent mechanism.4.TRIM29 regulated FN1 via mi R-873-5p in PTC cells.5.TRIM29 inhibited premi R-873 cleavage by suppressing recruiting Dicer and TRBP.6.Lnc RNA CYTOR was involved in regulation of FN1 by TRIM29.7.TRIM29 suppressed pre-mi R873 cleaving to generate mature mi RNAs via upregulation of CYTOR.Conclusion: TRIM29 inhibited mi R-873-5P biogenesis via lnc RNA CYTOR sponging premi R-873 to upregulate FN1 and promoted migration and invasion of papillary thyroid cancer cells. |
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