TRIM37 Promotes The Proliferation And Invasion By Regulating Wnt/β-catenin Signaling Pathway In Cervical Cancer Cells

Background and Purpose:Cervical cancer ranks first in malignant reproductive system tumors,which seriously endangers women’s health.At the beginning,about 50% of patients are diagnosed in advanced disease.In recent years radiotherapy;targeted therapy and immunotherapy have made great progress,the effective rate has not been significantly improved,and 50% of patients with advanced cervical cancer will have recurrence and metastasis,which greatly reduces the survival rate.Therefore,it is most important and urgency things to explore the molecular mechanism and to find new molecular targets related to invasion and metastasis of cervical cancer cells in order to improve the efficacy and prognosis of cervical cancer patients.The TRIM family is involved in a variety of physiological and pathological processes,including cell proliferation,autophagy innate immune inflammation,and tumorigenesis.Studies have confirmed that TRIM37 plays a cancer-promoting role in the development and progression of pancreatic cancer,liver cancer,breast cancer and intestinal cancer,but its biological function and mechanism in cervical cancer have not been clarified.The Wnt/β-catenin signaling pathway plays an important role in tumor genesis and development,including cell proliferation,invasion and metastasis,epithelial-mesenchymal transformation,and tumor stem cell characterization.So whether TRIM37 is associated with the Wnt/ β-catenin pathway,and whether TRIM37 can play a biological role by regulating the Wnt/ β-catenin pathway.Clarifying the mechanism of abnormal activation of this signaling pathway in cervical cancer will not only help to further understand the malignant invasion of cervical cancer and the proliferation mechanism of cancer cells,but also provide new ideas for the treatment of clinical diseases.The purpose of this study was to analyze the expression and function of TRIM37 in patients with cervical cancer.To explore the biological process of TRIM37 affecting malignant invasion of cervical cancer and proliferation of cancer cells;To investigate whether TRIM37 can influence the Wnt/β-catenin signaling pathway,so as to indirectly affect the metastasis and invasion process of cervical cancer cells.The aim is to provide reference for the clinical diagnosis and treatment of cervical cancer patients and improve the prognosis of patients.Part 1: Expression of TRIM37 in cervical cancer and clinical significanceObjective: Observe the expression of TRIM37 in cervical cancer tissue and explore its relationship with clinicopathological factors and prognosis of cervical cancer.Methods: The mRNA and protein expression levels of TRIM37 were detected by RT-PCR and Western-blot.CCK-8 and plate cloning experiments were carried out to detect the proliferation ability of cervical cancer cells after the collection of sample tissues.Transwell cell migration assay was conducted to understand cell migration,and cell invasion ability was analyzed by cell scratch assay.The changes of the marker proteins during EMT were studied by immunofluorescence method.Results:The expression of TRIM37 in cervical cancer tissues was significantly increased compared with the adjacent tissue.The results of chi-square test showed that: high expression of TRIM37 inversely correlated with and tumor diameter(P=0.005);Lymph node metastasis(P=0.001)and FIGO staging(P=0.000).Log-rank results:Overall survival time(OS)in the patients with highly expression of TRIM37 was significantly inferior to that of patients with lower expression(40 months VS.70months;P=0.001).COX regression analysis showed that the late FIGO stage,lymph node metastasis and high expression of TRIM37 were closely related to poor prognosis.Conclusion: The expression of TRIM37 in cervical cancer tissue was significantly increased.TRIM37 expression is closely related to late FIGO staging lymph node metastasis.High expression of TRIM37 affects the survival and prognosis of the patients with cervical cancer,indicating a poor prognosis.High expression of TRIM37 is an important prognosis with poor survival for the patients with cervical cancer.Part 2: The biological function and EMT of TRIM37 in cervical cancer cellsObjective: To observe the expression of TRIM37 in cells of cervical cancer and explore its biological function and EMT in cervical cancer.Methods: RT-PCR and Western-blot method was used to detect the mRNA and protein expression of TRIM37;Detection of proliferation of cervical cancer cells by CCK-8 and plate cloning experiments;cell scratch test and transwell cell migration test were used to observe the changes of cell migration and invasion ability;Immunofluorescence was used to detect the changes of labeled proteins during EMT.Results: Overexpression of TRIM37 increased the invasion ability of SIHA and Hela cells in cervical cancer,but the proliferation and invasion ability of Hela and SIHA cells decreased after silencing TRIM37.Immunofluorescence results suggest that after interference TRIM37,Vimentin expression levels were significantly reduced,and the E-cadherin level increased significantly.After the over expression TRIM37,the opposite result appeared.Conclusion: TRIM37 can promote the proliferation and invasion of cervical cancer cells.TRIM37 promotes EMT of cervical cancer cells,silencing TRIM37 inhibit the EMT of cervical cancer cells.Part 3: TRIM37 Regulated the Wnt/β-catenin signaling pathway in cervical cancerObjective: To observe the effect of TRIM37 expression on the expression of core components in Wnt/β-catenin signaling pathway in cervical cancer.To provide a new target of the treatment for cervical cancer.Methods: Western-blot and RT-PCR were used to detect the changes of corresponding indexes in Wnt/β-catenin signaling pathway in nude mice transplanted tumor;Using Western blot to explore the effect of transfection TRIM37 sh RNA on the expression level of protein in core members of Wnt/β-catenin signaling pathway in cervical cancer cells and RT-PCR detection of the expression level of mRNA.Results: The protein expressions of β-catenin and cyclin D1,the core components of Wnt/β-catenin pathway,were down-regulated in the tumor tissues of TRIM37 interference group.TRIM37,c-myc,β-catenin and cyclin D1 protein levels were significantly down-regulated in He La and SIHA cells transfected with TRIM37 sh RNA compared with those transfected with scramble sh RNA.Similarly,the expression of mRNA levels with TRIM37,c-Myc,FOXM1 and Cyclin D1 in He La and SIHA cervical cancer cells transfected with TRIM37 PLASMID were also significantly down-regulated.Conclusion: The expression of TRIM37 can affect the expression level of core members downstream of Wnt/β-catenin signaling pathway,TRIM37 maybe inhibit tumor growth by inhibiting Wnt/β-catenin signaling pathway.