| Study Background and SignificanceBladder cancer,with an incidence ranking only next to prostate cancer,is the second most common urogenital malignant tumor in the world.Nevertheless,in China,bladder cancer is currently the most common malignant tumor,whose morbidity has been in an increasing tendency in recent years.With a distinct epidemiological survey characteristic,most of bladder cancer's pathological types are urothelium carcinoma(90%above);while squamous-cell carcinoma,adenocarcinoma and other types of cancer are rare.Just like most malignant tumor,there are many factors that can influence the prognosis of bladder cancer,such as the TNM staging of cancer,degrees of histopathological differentiation,depth of invasion,numbers of metastatic lymph nodes,and the proportion of LN positivity,etc.Its invasion,metastasis and relapse are primary causes to deaths of patients with bladder cancer.Incipient symptoms of bladder cancer are often painless gross hematuria or microscopic hematuria.Patients are apt to overlook treatments to these problems.And if allowed to progress,these symptoms may gradually develop from tunica mucosa to muscular layer,and ultimately turn to distant metastasis,then a poor prognosis.Therefore,finding out causes of tumor recurrence and metastasis in the early stage,as well as exploring new effective treatments can be especially significant for clinical practices.CD4~+T cells are important components of the cellular immunity.Recent studies prove that CD4~+T cells are not only helpers of CD8+ cytotoxic T cells,but also act as a role to assist NK cells in killing tumor cells.Although certain progresses have been made in immune researches of bladder cancer,researches on the correlation between expressions of CD4~+T cell and its subtype content in bladder cancer tissues and bladder cancer prognosis are still rare at home and abroad.There are three parts in this study.The first part:to observe expressions of CD4~+T lymphocyte and Treg cells in 131 specimens of non-muscle invasive bladder cancer(NMIBC)tissues removed through transurethral bladder cancer electrocision,and to carry out relevant retrospective analyses on clinical and pathological factors,exploring expressions of CD4~+T lymphocyte and Treg cell in non-muscle invasive bladder cancer tissues and its correlation with prognosis.The second part:to observe expressions of CD4~+T lymphocyte and Treg cells in 97 specimens of muscle invasive bladder cancer(MIBC)tissues exploring its correlation with prognosis and to analyze the difference between expressions of CD4~+T lymphocyte and Treg cells before and after the neoadjuvant chemotherapy respectively,exploring influences of chemotherapy on expressions of CD4~+T lymphocyte and Treg cells,in order to provide theory gist for the selection of reasonable chemotherapy regimens in clinical practices.The third part:to detect the expression of Treg cells,tumor proliferation index(proliferation index,PI),DNA ploidy and cell apoptosis rate four indexes in 45 cases of fresh bladder cancer tissues by using flow cytometry,and to discusse the correlation between them,clarifing their role in tumor proliferation and prognosis in order to predict the occurrence of bladder cancer,development and diagnosis.Part ?Study Objectives:This project aims to study expressions of CD4~+T lymphocyte and Treg subtype in non-muscle invasive bladder cancer and to explore its correlation with prognosis.Study method:there are 131 patients with non-muscle invasive bladder cancer in the study,and we detect the distribution of CD4~+T lymphocyte and Treg cells in this 131 non-muscle invasive bladder cancer tissues via immunohistochemical staining,and evaluates overall survival(OS)and relapse free survival(RFS)using Kaplan-Meier analysis method and Cox proportional hazards regression model,in order to explore the correlation between expressions of CD4~+T lymphocyte and Treg cells in non-muscle invasive bladder cancer tissues and the prognosis,with a consideration of factors in clinical pathology.Study findings:discovered through immunohistochemical staining detection,the positive features of CD4~+T lymphocytes and Treg cells mainly exists in cell membrane;and theirs expression proportion in non-muscle invasive bladder cancer tissues and normal epithelial cells of bladder mucosa are differences.Based on the average number of CD4~+T lymphocyte and Treg cells counting data collected under Nikon Eclipse80i microscope(The average value=1,400times high resolution),Patients with non-muscle invasive bladder cancer are divided into two groups:low density group and high density group.According to the results of single factor variable analysis,the density of both CD4~+T lymphocytes and Treg cells and the overall survival are in an inversely proportional relationship(P<0.05),while the density of The two and the prognosis of relapse survival are unrelated(P<0.05);Multivariate analyses demonstrate that the density of tumor-infiltrating CD4~+T lymphocytes and Treg cells are also independent prognosis factor for the overall survival(P<0.05).Conclusions for Part ?:Higher proportioned expressions of CD4~+T lymphocytes and Treg cells indicates a shorter survival of patients with non-muscle invasive bladder cancer;for these patients,the higher proportioned expressions of CD4~+T lymphocytes and Treg cells may be a sign of potential poor prognosis.Part ?Study Objectives:to explore the expressions of CD4~+T lymphocytes and Treg cells in patients with muscle invasive bladder cancer and its correlation with adjuvant chemotherapy and prognosis.Study Method:We collected clinical and pathological data from 97 patients who have undergone the radical total cystectomy in our clinical practices;among these patients,37 were treated with neoadjuvant chemotherapy before the operation,and 40 were treated with adjuvant chemotherapy after the operation.And we tested the expressions of CD4~+T lymphocytes and Treg cells in bladder cancer tissues of the patients before and after the chemotherapy via immunohistochemical staining to analyze the correlation between the expression of CD4~+T lymphocytes,Treg cells and various clinicopathologic factors.Through survival analysis,we try to find independent factors that may influence the prognosis of patients with muscle invasive bladder cancer.Study Findings:discovered through immunohistochemical staining detection,the positive features of CD4~+T lymphocytes and Treg cells mainly exists in cell membrane with different expression levels.According to statistical analysis,the expression of CD4~+T lymphocytes and Treg cells are irrelevant to factors,such as the patients5 age,gender,tumor location and size,which are no difference in statistics(P>0.05).According to results of single factor variable analysis,the density of both CD4~+T lymphocytes and the overall survival are irrelevant correlation(P>0.U5),while all of the density of Treg cells and TNM staging and Pathology classification and chemotherapy are related to the Overall survival(P<0.05);Results of multi-factor survival analysis demonstrate that clinical staging,Treg cells density and the chemotherapy treatment are relevant to the prognosis of muscle invasive bladder cancer patients(P<0.05),while factors such as age,gender,tumor location and size are irrelevant to the prognosis of muscle invasive bladder cancer patients(P>0.05).In addition,the study also found that neoadjuvant chemotherapy can significantly reduce the Treg cells expressing(P<0.05)but no obvious difference for CD4~+T cells(P>0.05).Conclusions for Part ?:The density of Treg cells and active neoadjuvant chemotherapy before and after operations can be independent influencing factors of prognosis for muscle invasive bladder cancer patients.Neoadjuvant chemotherapy before the operation can reduce the expression of Treg cells in muscle invasive bladder cancer patients,and density difference of Treg cells can be one of the predictors for the effectiveness of chemotherapy.Part ?Study Objective:To investigate the correlation between the cell proliferation,apoptosis and the expression of Treg cells in patients with bladder cancer.Study Methods:We collected 45 clinical resection of bladder cancer tissue samples,by using flow cytometry to detect the expression of the Treg cells,tumor proliferation index(proliferation index,PI),DNA ploidy and cell apoptosis rate index,and discussed the correlation between them,so as to clear the role of judgment in the malignant degree and prognosis.Study findings:discovered through flow cytometry detection,there is high expression of bladder cancer in the group of aneuploid tumors(75.56%);the expression of Treg cells and DI are closely associated with the differentiation of bladder cancer and the clinical stage(P<0.05);and PI is closely related with ploidy types,degree of differentiation and clinical staging in the bladder cancer(P<0.05);apoptosis rate has a close correlation with ploidy types and clinical staging of bladder cancer(P<0.05).With the increased expression of Treg cells gradually,the value of DI and PI were increased,the DI and PI were positively correlated with the expression of Treg cells(r=0.567,r=0.843,P<0.05),and the apoptotic rate was negatively correlated with the expression of Treg cells(r =-0.8502,P<0.05).Conclusions for Part ?:The cell proliferation and apoptosis in bladder cancer were abnormal expressed and associated with Treg cells expression,suggesting that the expression of Treg cells plays an important role in the development,occurrence and prognosis of bladder cancer.Conclusions for this Project:the expression of CD4~+T lymphocytes and Treg cells can be detected in bladder cancer tissues;and the density of CD4~+T lymphocytes and Treg cells are of certain predictive value for clinical prognosis of non-muscle invasive bladder cancer patients:higher proportioned expression of CD4~+T lymphocytes and Treg cells may indicate worse clinical prognosis;and for patients with muscle invasive bladder cancer,correlation between the expression of CD4~+T lymphocytes and clinical prognosis is uncertain,but the high expression of Treg cells have poor prognosis in patients,according to the change of the density of Treg cells before and after chemotherapy which may be accepted as a effective predictors of evaluation of chemotherapy;the expression of Treg cells were correlated with cancer cell proliferation,apoptosis in bladder cancer tissues which were confirmed by flow cytometry,suggesting that the expression of Treg cells plays an important role in the development,occurrence and prognosis of bladder cancer. |
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