The Changes Of CD4~+T Cell Subsets In Peripheral Blood And Tissues Of Patients With Endometrial Carcinoma And Their Correlation With Subgroups

Endometrial carcinoma(EC)is one of three major malignant tumor in female reproductive system.Its development is a complex biological process involving multiple factors and multi-stage,of which 80%are caused by estrogen without progestin antagonistic stimulation as type I EC.Accumulation of the study found that obesity,high blood pressure,type 2 diabetes and polycystic ovary syndrome and Metabolic syndrome are the pathogenesis of endometrial cancer risk factors.However,to thorough understanding the biological behavior of the tumor,it is also necessary to acknowledge the micro-environment where the cancer cells live including immune cells(T cells,B cells,NK cells,etc.),cell factor and fibroblasts,in which CD4~+T lymphocyte mediated cellular immunity plays an important role in tumor immunity.With the in-depth study of tumor immune micro-environment,it has been found that tumor micro-environment is closely related to the occurrence and metastasis of various malignant tumors.However,the study of CD4~+T cells and their subsets in relation to EC is still at the initial stage.The dynamic immune balance between CD4~+T cell subsets is maintained by cytokines.And once the balance is broken,it will lead to inflammation,autoimmune diseases and tumor.However,it is still unknown whether there is an interaction among CD4~+T cell subsets,and whether the mechanisms by CD4~+T cell subsets can jointly promote EC progression.Therefore,this study will be divided into four parts to preliminarily investigate the expression of CD4~+T cell subsets in peripheral blood and tissues of patients with EC and their correlation with CD4~+T cell subsets.Methods(1)Collected the peripheral blood assay indexes and clinicopathological characteristics of 351 cases of EC patients,analyze the relationship between the peripheral blood immune inflammatory cell indexes and the clinicopathological characteristics and prognosis of patients with EC,and explore whether F-NLR score and F-PLR score can be used as the prognostic indicators of immune inflammation in EC patients.(2)Mass cytometry and q RT-PCR were used to investigate the changes in the proportion and related functions of CD4~+T cell subgroups in peripheral blood of patients in the normal control group,atypical hyperplasia group and EC group from the level of T cell surface markers and transcription factor levels.The correlation between the subsets of CD4~+T cells in peripheral blood was preliminarily investigated.(3)Mass cytometry and q RT-PCR were used to investigate the changes in the proportion and related functions of CD4~+T cell subgroups in tissues of patients in the normal control group,atypical hyperplasia group and EC group from the level of T cell surface markers and transcription factor levels.The correlation between the proportions of CD4~+T cell subsets in tissues was preliminarily investigated.(4)In vitro experiment CD4~+T cells were extracted,and Transwell co-culture model was constructed with EC cell lines to preliminarily investigate the effects of CD4~+T cells on the migration and invasion ability of EC cells.Results(1)High F-NLR score was more closely related to patients with EC of pathological type II,G3,III+IV;High F-PLR score was more closely related to EC patients aged?60 years,postmenopausal status,pathological type II,stage III+IV and cervical interstromal involvement.Multivariate analysis affecting OS and PFS in EC patients showed that type II EC,FIGO stage III+IV,high F-NLR score and high F-PLR score were independent risk factors for prognosis in EC patients.(2)The expression levels of Th1 cells and T-bet m RNAs in the peripheral blood of the EC group were significantly lower than that of the NC group,while the expression levels of Th2 cells and GATA3 m RNA were significantly higher in the EC group.The expression levels of Th17 cells and ROR?t m RNA in peripheral blood of the EC group were significantly higher than that of the NC group,and the expression levels of Treg cells and FOXP3 m RNA were also significantly higher.Meanwhile,the expression levels of Th22 cells and AHR m RNA in circulating peripheral blood of EC patients were significantly higher than those in NC group and AH group.What's more in the EC group,Th1 cells showed a positive correlation with Th2 cells,while in the NC group,Th17 cells and Treg cells showed a positive correlation.(3)The expression levels of Th1 cell and T-bet m RNA in the tissues of EC group and AH group were lower than those in the NC group.The proportion of Th2 cell in the EC group was significantly higher than that in the NC group,while the expression level of GATA3 m RNA was gradually increased in the AH group and the EC group.The proportion of Th17 cell,the expression level of ROR?t m RNA,the proportion of Treg cells,the expression level of FOXP3 m RNA,the expression level of Th22 cells and AHR m RNA in the EC group were all higher than those in the NC group.The results of mass cytometry showed a positive correlation between Th1 cell frequency and Th2 cell frequency in the EC group.(4)Transwell co-culture experiment showed that CD4~+T cells have a certain degree of influence on the invasion ability of Ishikawa cells and KLE cells in vitro.Conclusion(1)Patients with higher peripheral blood F-NLR and F-PLR scores before the initial treatment of EC may be independent risk factors for potentially poor prognosis of PFS and OS,and the new grading system combined with the immune inflammatory indicators of F-NLR and F-PLR can be used as a simple non-invasive prognostic indicator of EC.(2)EC patients Th1 in the peripheral blood and tissues and main transcription factor T-bet m RNA expression level significantly lower than NC group,Th2 and GATA3m RNA transcription factor expression level,Th17 and ROR gamma T m RNA transcription factors expression level,Treg and FOXP3 m RNA transcription factor expression level,Th22 and AHR m RNA transcription factor expression level significantly increased in the NC group.This suggests that in the peripheral blood and local tumor microenvironment Th1/Th2 proportional imbalance,the proportion of Th17/Treg imbalances and Th22 may be involved in the incidence of EC.(3)Transwell co-culture experiment showed that CD4~+T cells have a certain degree of influence on the invasion ability of Ishikawa cells and KLE cells in vitro.