Suppression Of The Smurf1 Expression Inhibits Tumor Progression In Gliomas

Objective:1.To investigate Smurfl expression in different grades glioma tissues and the correlation with migration marker E-cadherin and their clinicopathologic significance.Meanwhile,to discuss the role of Smurfl in the development and progression of gliomas.2.To study the expression of Smurfl in glioma cell lines and the effect of Smurfl on the invasion and apoptosis of glioma cell lines,and to further explore the molecular pathogenesis of glioma in order to provide a new target for its treatment.Methods:1.At the organizational level,western blot were used to analyze the expression levels of Smurfl in six glioma samples.Immunohistochemical analysis was used to detect the expression of Smurfl in the 98 glioma cases.According to clinicopathologic data of gliomas patients,we analyze the relation between Smurfl,patient's age,gender,tumor location,type of surgery,and tumor size in 98 glioma cases.98 cases of gliomas patients were followed up,the correlation between Smurfl and survival curve were detected using the Kaplan-Meier method.Univariate and multivariate analyses using the Cox's proportional hazards model was constructed(including the clinicopathological parameters mentioned before).2.At the cellular level,the expression of Smurfl in glioma cell lines was detected by Western blot,and the glioma cells with higher Smurfl expression were screened out.After using Smurfl small interfering plasmid on the U87MG cell line,Wound healing and Transwell assay was used to analyze the effect of Smurfl on glioma cell migration and invasion.Flow cytometry was used to analyze the effect of Smurfl on glioma cell apoptosis Western blot was used to detect the expression of cleaved caspase-3 and cleaved PARP.Results:1.Western blot and immunohistochemical analysis demonstrated that Smurfl expression was evaluated and its level was correlated with the grade of malignancy.Beside,little remarkable correlation was observed between Smurfl expression and patient's age,gender,tumor location,type of surgery or tumor size.Spearman rank correlation analysis showed a negative correlation between Smurfl and E-cadherin(y=0.568;P<0.01).Kaplan-Meier survival curves indicated that up-regulation of Smurfl was significantly correlated with a lower survival rate(P<0.01).Multivariate Cox regression analysis indicated that Smurfl(P=0.001),E-cadherin(P=0.024),grading(P=0.035)can be used as an independent indicator of glioma prognosis.2.The expression of Smurfl in normal glial cells and gliomas was detected by Western blot.The expression levels of Smurfl were as follows:U251MG,U87MG,U118,A172,HEB.In the U87MG cells,the expression of Smurfl was inhibited by small interfering plasmids,and the migration ability of the cells was significantly decreased by Wound healing and Transwell assay.The expression of E-cadherin was increased and the expression of Vimentin was decreased.3.In the U87MG cells,the expression of MDM2 was decreased,the expression of p53 was increased,the apoptosis rate increased,the expression of cleaved caspase-3 and the cleaved PARP were increasedConclusions:1.At the tissue level,Smurfl was highly expressed in glioma tissue and positively correlated with tumor grade,and was negatively correlated with E-cadherin expression,which was closely related to the poor prognosis of glioma patients,suggesting that Smurfl may be associated with the occurrence and development of glioma,can be used as a prognostic indicator of glioma.2.At the cellular level,The suppression of Smurfl expression by siRNA transfection was shown to significantly decreased the migration ability of glioma cells and increased the apoptosis.These data suggested that Smurfl could be a potential therapeutic target for gliomas.