The Function And Mechanism Of TRIM47 In The Development And Progression Of Non-small Cell Lung Cancer

Objective: Lung cancer is the most common malignancy with the highest morbidity and mortality in the world,which pose a great threat to human health.To investigate the molecular mechanism of invasion and metastasis in non-small cell lung cancer(NSCLC),and then blocking some of the key joint,it is an effective treatment and a hot research field.Some studies have shown that a variety of TRIM family proteins act as oncogenes and promote the proliferation and metastasis of cancer cells in the development of NSCLC.However,the exact function of TRIM47 in NSCLC is not fully understood.The aim of this study was to investigate the expression of TRIM47 and its role in the proliferation and metastasis of NSCLC,which will provide a new strategy and preliminary experimental basis for the treatment of NSCLC.Methods:(1)Immunohistochemistry and PCR were used to detect the expression of TRIM47 in tumor tissues and in normal adjacent tissues.The relationship between TRIM47 expression and various clinicopathological parameters was analyzed.Log-rank test was used to analyze the relationship between TRIM47 expression and prognosis,and Kaplan-Meier method was used to draw survival curve.COX proportional regression model was used to analyze the effect of clinicopathological parameters on prognosis of patients.To evaluate the potential capability of TRIM47 as a diagnostic biomarker for the prediction of patient survival,receiver operating characteristic(ROC)curves were conducted using TNM stage,TRIM47 level,or a combination of both.At the same time,the TCGA and GEO dataset were excavated to analyze the expression of TRIM47 in NSCLC and its relationship with the prognosis of patients.(2)Human TRIM47 vector was used to induce TRIM47 over-expressing in SPC-A1 cells,human TRIM47 shRNA vector was used to induce TRIM47-silence in A549?H358and H460 cells.A CCK-8 assay was used to evaluate the effects of TRIM47 on theproliferation of NSCLC cells.PI staining and flow cytometry analysis were used to evaluate the effects of TRIM47 knockdown on the cell cycle.The migration and invasion ability of NSCLC cells were assessed to investigate the function of TRIM47 in the metastasis of cancer cells.To verify the positive role of TRIM47 on tumorigenicity in vivo,we performed xenograft tumor assays using A549 cells stably infected by scramble shRNA control and TRIM47-shRNA lentiviruses.(3)High throughput RNA-sequencing data of the NSCLC cohort from TCGA and GEO were used to perform GSEA to probe the TRIM47-associated pathways in an unbiased manner.Western blot and qPCR were used to detect the expression of downstream genes in related signaling pathways,and the potential molecular mechanisms of TRIM47 was analyzed in NSCLC.Results:(1)TRIM47 expression level was significantly higher in tumor tissues than in normal adjacent tissues,and higher TRIM47 expression predicted poor relapse-free survival and overall survivall.TRIM47 expression was correlated with tumor size,tumor differentiation,TNM stage and lymph node metastasis.Multivariate Cox regression analyses showed that along with TNM stage and lymph node metastasis,overexpression of TRIM47(P = 0.017)could be considered an independent prognostic factor for NSCLC patients.The ROC curves analysis demonstrated that the combination of TRIM47 and TNM stage was superior to TRIM47 or TNM stage alone for the prediction of patient survival.(2)Overexpression of TRIM47 significantly enhanced the proliferation,invasion and migration ability of SPC-A1 cells.However,suppressing TRIM47 expression showed remarkable reduced the proliferation,migration and invasion ability in A549?H358 and H460 cells.We found that TRIM47 depletion induced G0/G1 phase arrest and inhibited cell proliferation in A549?H358 and H460 cells.TRIM47 silencing notably suppressed tumorigenicity in nude mice.(3)GSEA showed that high expression of TRIM47 was related to p53-cell cycle and NF?B-EMT signaling pathway,which were associated with proliferation and metastasis.Western blot analysis showed the expression of cell proliferation and metastasis relatedgenes were up-regulated in SPC-A1 cells,while we got the opposite results in A549?H358 and H460 cells.Conclusion: In our study,we systematically explored the expression level,biological functions and potential molecular mechanisms of TRIM47 in NSCLC.The results showed that overexpression of TRIM47 in tumor tissues predicted poor prognosis,and TRIM47 is one of the independent prognostic factor for NSCLC patients.TRIM47 may participate in the regulation of P53 and NF?B by promoting the ubiquitination and degradation of P53 and I?B,which advance the proliferation and metastasis of NSCLC.TRIM47 overexpression may serve as a useful prognostic factor and a potential treatment target for NSCLC.