The Study Of Expression, Clinical Significance Of TAMs In Bladder Carcinoma

Bladder cancer is one of the most common cancer in urothelial carcinoma, most of clinical pathology(over 90%) is transitional cell carcinoma in the most of country in the world, Non–muscle-invasive bladder cancer is treated by transurethral surgery, but part of cases will recur and metastasis within a short time, the followed pathological staging proved to be muscle-invasive cancer in some of them. Tumor microenvironment plays an important role during tumorigenesis, recurrence, metastasis, which involves a multiple steps and complicated processes. Tumor-associated macrophages(TAMs) have been described in many solid tumors, including bladder cancer, they make up a main proportion of the cellular elements in different tumors. Several preclinical as well as clinical studies have linked macrophages to tumor initiation, development, and metastasis. A high count of TAMs were associated with a poor prognosis, accelerated lymph angiogenesis, The present study was conducted to explore the expression, clinical significance of TAMs in bladder carcinoma.Part Ⅰ The value of M2 tumor associated macrophages in predicting first recurrence in non muscle-invasive bladder transitional cell cancer patientsObjective: Tumor associated macrophages(TAMs) enhance the development of malignant tumor. CD163+ macrophages is considered to be expressed exclusively on cells of the monocyte/macrophages lineages. The present study is to explore the releationship of the infiltration of TAMs and tumor pathology with first recurrence of non muscle-invasive bladder transitional cell cancer(NMIBC).Methods : Diagnostic tissue from 337 patients with NMIBC was evaluated retrospectively by immunohistochemical analysis for CD163. Clinical datas and patient characteristics include age, gender, tumor size, tumor grade, multiplicity and TAMs density. Their correlation between clinicopathological variables and patients outcome were statistically analyzed with SPSS 21.0 statistical package χ2 test, fisher exact t test and Cox regression model were used for multivariate analysis. Kaplan-Meier survival cure and log-rank test were used to compare the survival rate between different groups with α=0.05 as statistical standard.Results: We combined the mild infiltration TAMs and moderate infiltration TAMs into the group in tumor stroma, compared with severe infiltration group, the recurrence rates are 13.4%, 74.1%, respectively. The recurrence rates are 14.0 for tumor <3cm and 28.4% for tumor ≥3cm; 26.7% for multiple tumors and 14.9% for single tumor; 25.0% for G1 tumor, 17.7% for G2 tumor and 22.1% for G3 tumor, All statistically significant differences were observed. Among all patients, severe infiltration of TAMs in tumor stroma is closely correlated with high recurrence rate(P=0.006). whereas for recurrence only tumour grade, size and multiplicity were significantly independent predictors(P=0.008, 0.001, 0.046, respectively).Conclusions: TAMs is considered as a potential recurrence biomarkers in NMIBC, among the first recurrence risk factors of NMIBC, Tumor grade, tumor size, multiplicity are independent risk factors for non-muscle-invasive bladder cancer recurrence.Part Ⅱ To study the expression, clinical significance of TAMs, MVD and VEGF in patients with bladder cancerObjective: To explore the expression, clinical significance of CD68 labeled tumor associated macrophages(TAMs), CD34 labeled tumor-microvessel density(MVD) and VEGF in patients with bladder cancer.Methods: Immunohistochemistry assay were used to detect TAMs, MVD and VEGF in 153 cases of bladder cancer and 39 cases normal bladder tissues, the relation between TAMs and MVD, VEGF were studied. Their correlation between clinicopathological variables and the expression of CD68, CD34 and VEGF were statistically analyzed with SPSS 21.0 statistical package χ2 test, Spearman’s and Kendall’s test.Results: To compare with normal tissue, the infiltration densities of CD68 positive macrophages were significantly higher in bladder cancer tissue(P<0.01), the infiltration densities of CD68 in muscle invasive bladder cancer were also significantly higher in non-muscle invasive bladder cancer tissue(P<0.01). the positive rates of VEGF, MVD and pathological grades of bladder cancer was correlated to the infiltration densities of CD68 positive macrophages(r=0.333 P<0.01, r=0.414 P<0.01, r=0.205 P=0.006; respectively).Conclusions: the deep infiltration densities of CD68 positive macrophages was associated with muscle invasive bladder cancer, CD68 positive macrophages may be involved with progression of bladder cancer. CD68 positive macrophages, VEGF and tumor-MVD have close relationships with the biology behaviors of bladder transitional cell cancer, the infiltration densities of CD68 can be used to differentiate clinicopathological stages.Part Ⅲ The expression character of monocyte-macrophages in tumor microenvironment in patients with bladderl cancerObjective: To investigate the expression character of CD163+/CD206+ monocyte-macrophages in the microenvironment of bladder cancer.Methods:Fluorescence microscopy or flow cytometry were performed to observe the expression character of infiltrating cells CD45+CD14+CD163+ monocytes/macrophages in tumor tissue from patients with bladder cancer. A tumor-bearing mice model was constructed using the mouse bladder carcinoma cell line MB49 in C57/BL6 mice. The dynamic expression of CD11b+F4/80+CD206+ monocytes-macrophages were examined by flow cytometry at different time points of tumor-bearing in the model.Result: The levels of CD45+CD14+CD163+ monocyte-macrophage were significantly increased in tumor tissue compared with those in nontumor tissue. CD40 high is an important character of CD45+CD14+CD163+ monocyte-macrophage compared with CD45+CD14+CD163- monocyte-macrophage in both tumor and non tumor tissue form bladder cancer patients. In tumor bearing mice experiment, it is found that levels of CD11b+F4/80+CD206+ monocyte-macrophage significantly increased with the tumor growth.Conclusion: CD163+/CD206+monocyte-macrophage show a significantly higher level in tumor microenvironment, and it is closely related to the progression of tumor. More important, CD40 high is found to be an important character of CD45+CD14+CD163+ monocyte-macrophage tumor microenvironment of bladder cancer.