Biological Functions Of TRIM37 In Nsclc And The Underlying Molecular Mechanisms

Objective:Lung cancer is the most prevalent cancer and remains the leading cause of cancer-related mortality worldwide.In clinical more than 80% patients are diagnosed with non-small cell lung carcinomas(NSCLC)histologically.One of the important reasons for the high mortality rate of lung cancer patients is that most of the patients have been diagnosed at intermediate to advanced stages when they has been lost the chance of offering and lost the chance of curative surgical treatment.Athough the applications in clinical treatment of NSCLC has made great progress currently,including radiotherapy,chemotherapy and surgery,Five year survival rate of patients with lung cancer is generally not very optimistic.Consequently,it is of great significance to find a new diagnostic and prognostic marker for early diagnosis and early treatment of lung cancer,which greatly contributes to distinguish the clinical efficacy of different patients and further individualize treatment.TRIM37,as a RING-B-box-coiled-coil(RBCC)subfamily protein,contains a RING finger and B box domain,which indicates it has the function of E3 ubiquitin ligases.The expression of TRIM37 was found to be associated with a variety of tumors,including colorectal cancer,pancreatic cancer and hepatocellular carcinoma.However,the clinical significance of TRIM37 in lung cancer,as well as the role of TRIM37 in the molecular mechanism of carcinogenesis and development of lung cancer,still remains to be elucidated.The results of our previous study showed that the expression of TRIM37 was over-expressed in NSCLC,and its expression level was related to the clinicopathological grade of nsclc and the survival and prognosis of the patients.The analysis of Cox proportional risk model showed that TRIM37 could be used as an independent factor to judge the survival and prognosis of patients.However,the role of TRIM37 in the development of NSCLC and its molecular mechanism need to be further explored.Therefore,this study further studies the biological function and molecular mechanism of TRIM37 in NSCLC.It is of great clinical significance to search for the specific prognostic markers of lung cancer and to improve the survival and prognosis of patients with lung cancer.Method:MTT assay and soft agar assays were used to examine the ability of TRIM37 to promote the proliferation of lung cancer cells.TUNEL and Annexin V assays were used to examine whether TRIM37 can promote the anti apoptotic ability of lung cancer cells.Chicken chorioallantoic membrane(CAM),tubule formation by HUVEC cells and transwell assays were used to examine whether TRIM37 can promote angiogenesis in lung cancer cells.The model of nude mice was carried out and the tumor sections were further made into sections.The immunohistochemical staining of CD34 with tunel was used to examine whether TRIM37 could promote the proliferation,anti-apoptosis and angiogenesis of lung cancer cells in vivo.RT-PCR,luciferase assay and western blotting were used to examine whether TRIM37 activated the NF-κB signaling pathway in NSCLC.By transfection of mutated IκB mutant plasmid into TRIM37 over-expressed lung cancer cells,tunel assay,combine annexin v assays,Chicken chorioallantoic membrane(CAM),tubule formation by HUVEC cells and transwell assays were used to examine whether the biological functionof TRIM37 promotes proliferation,anti-apoptosis and angiogenesis of lung cancer cells by activating NF-κB signaling pathway.Result:1.By MTT assay and soft agar assays,we found that compared with the control group,the proliferation ability of lung cancer cells with high expression of TRIM37 was increased,but the proliferation ability of lung cancer cells with silencing expression of TRIM37 was decreased.2.By tunel assay and annexin v assay,we found that after cisplatin treatment,the anti-apoptotic ability of lung cancer cells with high expression of TRIM37 was enhanced,while the anti-apoptosis ability of lung cancer cells was decreased after treatment with cisplatin.3.By chicken chorioallantoic membrane(CAM),tubule formation by HUVEC cells and transwell assays,we found that high expression of TRIM37 increased the ability of promoting angiogenesis in lung cancer cells,while silencing expression of TRIM37 cells decreased the ability of promoting angiogenesis.4.By the model of nude mice,tunel assay of tumor sections of nude mice and immunohistochemical staining of CD34,we found that the high expression of trim37 could promote the proliferation of lung cancer cells in vivo,inhibit apoptosis and promote angiogenesis.5.By luciferase assay and Western blotting,we found that TRIM37 activated the NF-κB signaling pathway in NSCLC and promoted the transcription of the downstream gene of NF-κB signaling pathway.6.We firstly transfected the mutated IκBα mutant plasmid into TRIM37over-expressed lung cancer cells.By the tunel assay annexin v assay,chicken chorioallantoic membrane(CAM),tubule formation by HUVEC cells and transwell assaysshowed,we found that overexpression of TRIM37 promoted the proliferation of lung cancer cells,and the ability of anti-apoptosis and angiogenesis through activation of NF-κB signaling pathway.Conclusion:1.TRIM37 is associated with the development of NSCLC,which can promote the proliferation,anti-apoptosis and angiogenesis of lung cancer cells.2.TRIM37 can activate the NF-κB signaling pathway in NSCLC and promote the transcription of NF-κB signaling downstream gene.3.Overexpression of TRIM37 promotes the proliferation of lung cancer cells,and its anti-apoptotic and angiogenic effects are mainly through activation of NF-κB signaling pathway.