| ObjectiveEvaluate the expression of autophagy related factors LC3 and the number of CD68+ microglia,CD4+ T lymphocytes and CD8+ T lymphocytes in glioma tissues,investigate the relationship between the expression of autophagy related factors LC3 and the numbers of CD68+ microglia,CD4+ T lymphocytes and CD8+ T lymphocytes,as well as the clinical significance of those cells in gliomas.MethodsThe study group consisted of 68 patients with gliomas who were admitted to the Department of Pathology,Binzhou Medical University Hospital,China,Which were 43 paraffin-embedded glioma specimens from January 2010 to January 2013 and 25 fresh surgical specimens from January 2016 to January 2018,normal brain tissue that must be removed from the surgical approach as a control,fresh glioma specimens and normal brain tissues in liquid nitrogen were used for protein preparation.All patients with glioma underwent surgical treatment in the Department of Neurosurgery,Binzhou Medical University Hospital,China.Before surgery,no patient had been treated with radiotherapy,chemotherapy or combined radiotherapy and chemotherapy,and all patients were free of systemic immune disease and other serious chronic diseases.All patients were suffering from objective causes and only received one cycle of conventional chemotherapy.All cases were followed up by a return visit to the center or telephone.Immunohistochemistry,Real-time PCR and Western Blotting were used to detect the expression of autophagy related factors LC3 in glioma tissues and in normal brain tissues,and the number of CD68+ microglia,CD4+ T lymphocytes and CD8+ T lymphocytes were detected by immunohistochemistry,Spearman correlation analysis was used to analyze the association of autophagy related factors LC3 with CD68+ microglia,CD4+ T lymphocytes and CD8+ T lymphocytes,Chi-square test analysis of the relationship between factors and clinicopathological factors,Cox proportional hazards regression model analysis of prognostic factors in patients with glioma.Results1.Immunohistochemistry,Real-time PCR and Western Blotting showed that The expression of autophagy related factors LC3 in normal brain tissue,low-grade gliomas,and high-grade gliomas are elevated to varying degrees(P<0.01).2.Immunohistochemical detection showed that the numbers of CD68+ microglia,CD4+ T lymphocytes and CD8+ T lymphocytes in gliomas was higher than that in normal brain tissues(P<0.01),and the high-grade gliomas were higher than those in low-grade gliomas(P<0.01);3.The results of Spearman correlation showed that the expression of autophagy related factors LC3 was positively correlated with the numbers of CD68+ microglia,CD4+ T lymphocytes,and CD8+ T lymphocytes(P<0.05).4.Chi square test showed that the expression of autophagy related factor LC3 and the infiltrated number of CD68+ microglia,CD4+ T lymphocytes and CD8+T lymphocytes were all related to the grade of glioma(P<0.05),and the higher the tumor grade,the higher the expression/infiltration.5.Furthermore,in low-grade gliomas,the expression of autophagy-associated factor LC3 and the infiltration of CD68+ microglia,CD4+ T lymphocytes,and CD8+T lymphocytes are closely related to the prognosis of patients.Detection of infiltration and the expression of LC3,CD68+ microglial,CD4+ T lymphocytes and CD8+T lymphocytes has an important reference value for evaluation the prognosis of the patients with low-grade gliomas.(P>0.05).Conclusion1.The autophagic level of gliomas and the infiltration of CD68+ microglia,CD4+ T lymphocytes,and CD8+ T lymphocytes are all related to pathological grade.2.The expression of autophagy related factors LC3 is positively correlated with the number of CD68+ microglia,CD4+ T lymphocytes and CD8+T lymphocytes,suggesting that the change of autophagy level may be one of the factors that influence the cellular immune function in glioma.3.In low-grade gliomas,the 5-year survival rates of autophagy related factors LC3,CD68+ microglia,CD4+ T lymphocyte and CD8+ T lymphocyte positive group and negative group were statistically significant,detection of the expression of autophagy related factors LC3 and the number of CD68+ microglia and CD8+T lymphocytes has important reference value for judging the prognosis of patients with glioma. |
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