SENP1 Expression In Neuroblastoma And Its Regulation Of Tumor Cell Migration And Invasion

Neuroblastoma is one of the most common lethal malignant solid tumors in children.It has high malignancy,strong invasiveness,and early distant metastasis.Although it has undergone comprehensive treatment such as surgery,chemotherapy,radiotherapy and even bone marrow transplantation,There is still a high recurrence rate,and the long-term survival rate is less than 40%.Tumor recurrence and metastasis are one of the main causes of death in children.Preventing and reducing metastasis is the main challenge for curing neuroblastoma and preventing recurrence.However,the molecular mechanism of the occurrence and development of neuroblastoma invasion and metastasis is still unclear.Therefore,exploring the invasion and metastasis mechanism of neuroblastoma and looking for possible intervention measures are of great significance for improving the quality of life of patients and the tumor-free survival rate.The invasion and metastasis of neuroblastoma involves a series of complex processes and is regulated by a variety of cytokines,and these factors are often regulated by protein post-translational modifications.The post-translational modification of protein is a key signal pathway for protein function regulation.Small ubiquitin-like modification(SUMOylation)is a newly discovered important form of post-translational modification in cells.Modifications can participate in transcription regulation,nuclear transport,maintenance of genome integrity,and signal transduction.Inner activity is an important multifunctional way of protein post-translational modification.The dysregulation of SUMO modification function may lead to the occurrence of certain diseases.Protein SUMO modification is a dynamic and reversible process.The important determinant for the SUMO modification of proteins is the deSUMO process mediated by the small ubiquitin-related modifier specific protease 1,SENP1,which maintains one of the proteins required for normal cell physiology.The balance between them plays a key role in cell regulation.The abnormal expression of SENP1 is closely related to the occurrence and development of many tumors.However,the role of SENP1 in neuroblastoma and its mechanism are currently poorly understood.In this study,through the systematic study of neuroblastoma tumor specimens and tumor cell lines,the expression level of SENP1 in human neuroblastoma specimens was observed,and SENP1 was over expressed or silenced in neuroblastoma cell lines in order to explore SENP1 The role in the development of neuroblastoma,to further clarify the mechanism of SENP1 in neuroblastoma migration and invasion.Part 1The clinical features of neuroblastoma and the expression of SENP1 in neuroblastoma tissuesObjective:To review and summarize the clinical characteristics of neuroblastoma surgical cases admitted to our hospital in the past five years,and to detect the expression level of SENP1 in the primary and metastatic tissues of neuroblastoma.Methods:Collected and analyzed the data of neuroblastoma cases who were treated in the surgical department in Children’s Hospital of Soochow University from December 2015 to December 2020.All cases underwent tumor resection,and postoperative pathological biopsy confirmed neuroblastoma.RT-PCR analysis was used to detect the expression of SENP1 in the primary and metastatiues of neuroblastoma,and the expression of SENP1 in the primary and metastatic tissues of neuroblastoma was analyzed by western blotting.Results:Completely collected clinical data of 69 cases,including 41 males(58.9%)and 28 females(41.1%).Age at first visit from 5 days to 9 years and 5 months.Average age at first visit is 3 years and 2 months.The median follow-up time was 22.5 months(3-63 months).The primary tumor was located in the adrenal gland in 24 cases(34.8%),in the posterior peritoneum in 20 cases(29.0%),in the mediastinum in 19 cases(27.5%),in other parts,including the neck,spinal canal,and sacral coccyx in 6 cases.(8.7%).There were 7 cases(10.1%)in stage 1 of children with INSS,13 cases in stage 2(18.9%),24 cases in stage 3(34.8%),and 25 cases in stage 4(36.2%).39 cases of neuroblastoma(56.5%),ganglion cell neuroblastoma,22 cases(31.9%)of mixed type,7 cases of ganglion cell neuroma(10.1%),ganglion cell neuroblastoma,1 case of nodular type(1.5%).The efficacy was compared according to different INSS stages,different primary sites and different age groups.The results showed that the efficacy of children with INSS stages 3 and 4 was significantly worse than that of stage 1.2(P<0.05).The posterior peritoneum(including the adrenal gland),mediastinum and Other parts(including neck,spinal canal,pelvis,etc.)have no difference in curative effect between different primary parts(P>0.05).The curative effect of children aged 1.5 to 5 years is better than that of children less than 1.5 years of age(P<0.05)).The expression rate of SENP1 mRNA and protein in neuroblastoma tumor tissue was 100%,and the expression level of SENP1 mRNA in neuroblastoma metastasis tissue was significantly higher than that of matched primary site(P<0.05).Neuroblastoma metastasis tissue The expression level of SENP1 protein was significantly higher than the matched primary site(P<0.05).Conclusion:Neuroblastoma has diverse clinical behaviors,early distant metastasis,and poor clinical efficacy in high-risk children.SENP1 is widely expressed in neuroblastoma,and SENP1 is overexpressed in metastatic neuroblastoma tissue.Part 2Regulation of SENP1 on tumor migration and invasion in neuroblastoma.Objective:To investigate the regulation of SENP1 on neuroblastoma cell migration and invasion.Methods:The Flag-SENP1 eukaryotic expression plasmid was transfected into neuroblastoma cells through liposome mediation,so that SENP1 was highly expressed in the cells,and then the BrdU experiment was used to detect the proliferation of the cells.Transwell Experiments and scratch experiments were used to detect the migration and invasion ability of cells,and the expression levels of migration and invasion related proteins CDH1,MMP2 and MMP9 were detected by RT-PCR and Western blot.On the contrary,design siRNA targeting SENP1 to silence SENP1,then use BrdU experiment to detect cell proliferation,Transwell experiment to detect cell migration and invasion ability,and RT-PCR and Western blot to detect the migration and invasion related proteins CDH1,MMP2 and MMP9 The expression level.Results:After liposome-mediated transfection of the Flag-SENP1 eukaryotic expression plasmid into tumor cells in neuroblastoma cells,SENP1 was highly expressed in the cells,and the proliferation ability of tumor cells was slightly enhanced(P<0.05);The migration and invasion ability of cells was significantly enhanced(P<0.05);the expression of invasion-related protein CDH1 was partially decreased(P<0.05),and the expression of MMP9 and MMP2 was partially increased(P<0.05).On the contrary,when SENP1 is silenced by siRNA targeting SENP1,the expression of SENP1 in the cell is inhibited,and the proliferation ability of tumor cells is slightly reduced(P<0.05);the migration and invasion ability of tumor cells is significantly reduced(P<0.05);invasion-related proteins The expression of CDH1 was partially increased(P<0.05),and the expression of MMP9 and MMP2 was partially decreased(P<0.05).Conclusion:SENP1 is closely related to many aspects of neuroblastoma cell proliferation,migration and invasion,and it may act by regulating the migration and invasion-related proteins CDH1,MMP2 and MMP9.