Expression Significance Of CDKN2A Probe Combined With Smo And GLi1 Genes In Malignant Mesothelioma

Objective:To investigate the differential expression of CDKN2 A gene detection probe combined with Smo and GLi1 genes in Hedgehog pathway in malignant mesothelioma specimens,and the relationship between the gene expression and the immunoinfiltrating cell of mesothelioma.Effusion specimen detection is less traumatic and easy to obtain for patients.This method has not been reported in China at present.This study attempts to provide a new method and idea for further exploring the diagnosis and development of mesothelioma.Methods:1.CDKN2 A expression in 130 cases of malignant mesothelioma and 50 cases of benign mesothelioma(serous cavity effusion specimens,biopsy specimens)was detected by fluorescence in situ hybridization.The protein expression levels of Smo and GLi1 in malignant mesothelioma tissues were detected by immunohistochemistry to analyze the clinicopathologic significance of Smo and GLi1 protein expression in malignant mesothelioma tissues.The m RNA expressions of Smo and GLi1 genes in patients with malignant mesothelioma and benign mesothelioma were detected by reverse transcription polymerase chain reaction method,and P<0.05 was considered statistically significant.2.Patients with malignant mesothelioma were followed up for statistical analysis of Smo and GLi1 gene protein expression levels and patients’ survival,and survival curves were drawn.Cox proportional hazard regression analysis was used to establish a proportional hazard model for multivariate survival analysis data,and P<0.05 was considered statistically significant.3.GEO database was used to screen out immune cells closely related to the molecular mechanism and development of malignant mesothelioma through data set GSE51024.TIMER database was used to further explore the relationship between immune cell infiltration in mesothelioma,Smo and GLi1 genes,and survival prognosis of patients.Results:1.Fluorescence in situ hybridization showed that CDKN2 A gene fragment was absent in malignant mesothelioma tissue.Immunohistochemistry showed that the expression level of Smo and GLi1 genes in malignant mesothelioma was higher than that in benign mesothelioma.2.In malignant mesothelioma tissue,CDKN2 A gene probe combined with immunohistochemical Smo and GLi1 gene were used to detect the results of serous cavity effusion specimen and biopsy specimen.There was no significant difference between the two detection methods(P>0.05),and the results of the two samples were highly consistent.There was a positive correlation between Smo protein expression and GLi1 protein expression in malignant mesothelioma tissue(r=0.673,P<0.05).3.The results of reverse transcription polymerase chain reaction hybridization showed that the m RNA expression levels of Smo and GLi1 in malignant mesothelioma tissues were higher than those in benign mesothelioma tissues,and the expression levels were significantly different(P<0.05).4.The expressions of Smo and GLi1 protein were correlated with age,tumor site,asbestos exposure history and P53 expression in patients with malignant mesothelioma(P<0.05).The expression levels of Smo and GLi1 genes were negatively correlated with good prognosis of patients with mesothelioma(P<0.05).The higher the expression levels,the worse the survival prognosis of patients with mesothelioma.Cox proportional risk model indicated that tumor location,patient sex,asbestos exposure history and protein expression of Smo and GLi1 genes were independent risk factors for prognosis in patients with mesothelioma.5.GEO database screening and TIMER database analysis showed that the infiltration of immune cells in malignant mesothelioma was closely related to the expressions of Smo and GLi1 genes.The degree of immune cell infiltration was closely correlated with the prognosis of patients with malignant mesothelioma(P<0.05).Conclusions:The expression of CDKN2 A gene fragment in malignant mesothelioma tissue was significantly decreased,and the expression levels of Smo and GLi1 protein were higher than those in benign mesothelioma tissue,and the m RNA expression trend was consistent with the protein expression level.The expression of Smo and GLi1 genes is closely related to the site of malignant mesothelioma,asbestos exposure history and survival prognosis of patients.Gender,asbestos exposure history,tumor site and Smo and GLi1 gene expression levels are independent prognostic risk factors for mesothelioma patients.The degree of immune cell infiltration is closely related to the expression of Smo and GLi1 genes and survival prognosis of patients with mesothelioma.The use of malignant mesothelioma serous cavity effusion specimens for fluorescence in situ hybridization combined with immunohistochemical detection can greatly reduce the pain of patients,and can be used as a new method for screening malignant mesothelioma in early and middle clinical stages.