| Objective:Thyroid cancer(TC)is the most common malignant tumor of human endocrine system,accounting for about 1% of systemic malignant tumors.Its incidence is increasing year by year,and it is one of the fastest growing tumors in recent years.Although most patients with thyroid cancer with follicular cell origin have a good prognosis,conventional treatment(excision combined with radioactive iodine ablation and thyroid hormone therapy for TSH)can cause refractory concurrency in10%-15% of these patients.Symptoms,and metastatic follicular cell-derived thyroid cancer patients are ineffective for chemotherapy and external exposure,with a10-year survival of only 40-50%.In addition,thyroid undifferentiated cancer is more deadly,and there is currently no effective systemic treatment,which seriously endangers human health.Therefore,understanding the molecular mechanism of TC is particularly important.This study mainly describes the molecular mechanism of TC by analyzing the expression of TRIP13 in thyroid carcinoma and its correlation with clinicopathological parameters of thyroid cancer from the clinical perspective.After silencing the TRIP13 gene,the thyroid cancer cell viability,cycle,and withering were observed.The effects of death and invasion and migration ability;analysis of the regulation and mechanism of TRIP13 on the downstream protein TTC5/p53,and finally the effect of TRIP13 on the biological behavior of thyroid cancer cells through TTC5/p53 signaling pathway..Method:1.Collect 100 surgically removed samples of thyroid cancer tissues and adjacent normal tissues were collected,and mRNA expression levels of TRIP13,TTC5/p53,e-cadherin and n-cadherin were detected by real-time quantitative PCR(qRT-PCR).Immunohistochemistry was used to detect the expression levels of TRIP13,TTC5/p53,e-cadherin,n-cadherin and other proteins in thyroid cancer tissues and adjacent tissues.TRIP13 mRNA expression levels of different thyroid cancer cell lineswere detected by real-time quantitative PCR,and TRIP13 protein expression levels of different thyroid cancer cell lines were detected by Western blot..2.Build and screen the TRIP13 interference fragment,detect the interference effect by real-time fluorescent quantitative PCR and Western blot;screen the thyroid cancer cells after screening the best interference fragment,and examine the effect of CCK8 on the thyroid cancer cell viability after silencing TRIP13;The effects of silencing TRIP13 on the cell cycle and apoptosis of thyroid cancer cells were detected.The cell scratch test and Transwell assay were used to detect the invasion and migration of thyroid cancer cells after silencing TRIP13.3.Real-time quantitative PCR(qRT-PCR)was used to detect the changes of TTC5/p53,E-cadherin and N-cadherin mRNA after silencing TRIP13.The changes of TTC5/p53,E-cadherin and N-cadherin protein after silencing TRIP13 were detected by Western blot.It is clear that TRIP13 affects the biological function of thyroid cancer cells by regulating the TTC5/p53 signaling pathway.Result:1.Compared with normal thyroid tissues,TRIP13 and n-cadherin protein and mRNA expression levels in thyroid cancer tissues were significantly increased,while TTC5,p-p5 and e-cadherin protein and mRNA expression levels were significantly decreased,with statistically significant differences(p < 0.05).In addition,in the three thyroid cancer cell lines C643,BHT101 and TPC1,qRT-PCR and Western blot results showed that TRIP13 mRNA and protein expression levels in TPC1 and BHT101 were significantly higher than those in C643 thyroid cancer cells,with statistically significant differences.Therefore,TPC1 and BHT101 were selected for subsequent interference experiments(P < 0.05).2.After interfering with the expression of TRIP13 in the TPC1 and BHT101 cell lines,the CCK8 results showed that the cell viability of the 24 h and 48 h interference groups decreased significantly compared with the control group;the flow cytometry results showed that after interference with TRIP13 The period was significantly increased,the G2 phase was significantly decreased,and the apoptosis was also significantly increased.The difference was statistically significant(P<0.05).The cellscratch and Transwell results showed that the invasion and migration ability of thyroid cancer cells decreased significantly after interference with TRIP13.The difference was statistically significant(P < 0.05).3.In TPC1 and BHT101 cell lines,the expression of TTC5 and E-cadherin was significantly increased after interfering with TRIP13,the expression of N-cadherin was significantly decreased,and the expression of p-p53 was significantly increased in the TPC1 and BHT101 cell lines.The difference was statistically significant.(P<0.05).Conclusion:TRIP13 is highly expressed in thyroid cancer tissues and cells.TRIP13 affects the invasion,migration,cycle and apoptosis of thyroid cancer through TTC5/p53 signaling pathway,providing a new target for molecular therapy of thyroid cancer. |
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