Effects Of TRIM59 On Proliferation, Apoptosis, And EMT Of Human Nasopharyngeal Carcinoma Cells

Objective: Most nasopharyngeal carcinoma patients are not easily diagnosed in the early stage and are only discovered in the middle to late stages of the cancer.Therefore,although nasopharyngeal carcinoma patients are sensitive to radiotherapy,the 5-year survival rate of advanced nasopharyngeal carcinoma patients is still relatively low.TRIM59 is associated with tumor cell proliferation,apoptosis,and EMT.This study aims to explore the effect of TRIM59 on the malignant progression of nasopharyngeal carcinoma cells,thereby providing potential targets for targeted treatment of nasopharyngeal carcinoma.Methods: 1.Database was queried for TRIM59 expression in head and neck squamous carcinoma(HNSC).western blot and q RT-PCR were performed to detect TRIM59 expression levels in NPC cell lines(HNE1 and CNE-2Z)and human normal nasal mucosal epithelial cell lines(HNEp C).2.TRIM59 knockdown groups were performed in HNE1 and CNE-2Z cells(si-TRIM59)and TRIM59 overexpression group(TRIM59-mimics)and the corresponding negative control group(si-NC/mimic-NC)were constructed.3.CCK8 and cell cloning assays to analyze the effect of TRIM59 on cell proliferation.Annexin V-FITC/PI double-staining assay to analyze the effect of TRIM59 on cell apoptosis.Analysis of the effect of TRIM59 on cell EMT ability by cell scratch and Transwell test.4.Western blot to analyze the effect of TRIM59 on EMT and apoptosis-related molecular mechanism of NPC cells.5.The effects of TRIM59 on tumor growth in nude mice were analyzed.q RT-PCR was used to detect the expression level of TRIM59 in the tumor,and Western blot was used to detect the expression level of EMTrelated proteins in the tumor.6.Western blot analysis of the effect of TRIM59 on p53 protein.Results: 1.Database search revealed that TRIM59 expression was elevated in HNSC,and Western blot and q RT-PCR confirmed that TRIM59 expression was higher in NPC cells than in human normal nasal mucosal epithelial cell lines(P<0.05).2.Western blot verified transfection efficiency,in which TRIM59 expression in the si-TRIM59 group was lower than that of the si-NC group,while TRIM59 expression in the TRIM59-mimics group was higher than that of the mimic-NC group(P<0.05),and subsequent experiments were performed on this basis.3.CCK8 and cell cloning experiments confirmed that proliferation was inhibited in the si-TRIM59 group,while proliferation was promoted in the TRIM59-mimics group(P<0.05).4.Annexin V-FITC/PI double staining method confirmed that the apoptosis rate increased in the si-TRIM59 group and decreased in the TRIM59-mimics group(P<0.05).Western blot found that Bcl-2 decreased and Bax increased in the si-TRIM59 group,while the TRIM59-mimics group showed the opposite result(P<0.05).5.Transwell and cell scratch assays revealed that the ability of EMT was inhibited in the si-TRIM59 group and enhanced in the TRIM59-mimics group;Western blot detection of proteins associated with EMT revealed that the si-TRIM59 group expression level of E-cadherin was up-regulated,while N-cadherin and Vimentin expression level were down-regulated,and the opposite result appeared in the TRIM59-mimics group(P<0.05).6.The results of the nude mouse allogeneic transplantation tumor experiment showed that the growth of tumor body could be inhibited after knocking down TRIM59,and the growth of tumor body could be promoted after upregulation TRIM59(P<0.05),q RT-PCR confirmed that the expression level of TRIM59 decreased in the si-TRIM59 group and increased in the TRIM59 group,and Western blot confirmed that the expression level of E-cadherin was upregulated after knocking down TRIM59,while the expression level of N-cadherin and Vimentin was down,and the opposite result was achieved after upregulation of TRIM59(P<0.05)7.Knockdown of TRIM59 would result in upregulation of p53 expression,while upregulation of TRIM59 resulted in p53 expression down-regulated(P<0.05).Conclusion: Elevated expression of TRIM59 in NPC cells promotes malignant behavior of NPC cells and also regulates p53 protein expression levels.Therefore,the exploration of TRIM59 may provide a new theoretical basis for the treatment of NPC.