The Significance Of CCR7 Expression In Bladder Urothelial Carcinoma

Part one The correlation between expression of CCR7 in bladder urothelial carcinoma (BUC) and lymphatic node metastasisAim Explore the situation of expression of CCR7 in BUC, which relationship whit lymphatic node metastasis is also investigated, and evaluate the correlation between CCR7 with the parameters of BUC such as tumor stage and grade.Method The expression level of CCR7 in BUC was estimated by Immunohistochemistry technique, and the correlation between CCR7 with lymphatic node metastasis and other parameters of BUC was analyzed using statistic tools. And the influence factors were also tested.Result The positive rate of CCR7 expression among 57 patients was 82.5%, which was distinct higher than that in normal bladder tissue (20%); and the positive rate of CCR7 in lymphatic node metastasis group was 95.5%, distinct higher than that in none lymphatic node metastasis group (74.3%); The expression level of CCR7 was obvious greater in high stage group (T3-T4) than that in low stage group (T1-T2) (91.2%vs. 69.9%); The expression level of CCR7 was obvious greater in high grade group (G3) than that in low grade group (G1-G2) (87.5%vs.76.0%), and the expressions of CCR7 were no statistic difference with other clinic parameters; CCR7 expression, tumor stage and tumor grade were connected with lymphatic node status, but only the first was an independent one.Conclusion The expression of CCR7 was up-regulated in BUC, which is a positive independent influence factor of lymphatic node metastasis. Part two The influence of CCL21/CCR7 axis on T24 cells'biologic behaviorAim Estimate the influence of CCL21/CCR7 axis on T24 cells' biologic behavior such as migration, invasion, proliferation and anti-apoptosis.Method The T24 cell was grouped depend on the concentration of CCL21 treatment (0,50,100,200ng/mL). Transwell analysis was used to detect the effect of CCL21 to T24 migration and invasion; MTT was employed to evaluate the impact of CCL21 to T24 proliferation while FCM was employed to evaluate the impact of CCL 21 to T24 with anti-apoptosis ability to the apoptosis induced by ADM. What's more, the invasion associated protein MMP-2 and MMP-9, the apoptosis associated protein Bcl-2 and Bax was detected by western blot. Added, CCR7 antibody was used to evaluate the role of CCR7 in CCL21 usage.Result The T24 cells showed more powerful progresses of migration and invasion under the treatment of CCL21, and the affect was depend on the concentration of CCL21. CCL21 would up-regulate the expression of MMP-2 and MMP-9 proteins. Otherwise, CCL21 could promote the proliferation and anti-apoptosis ability of T24. The latter might associated with the up-regulation of Bcl-2 protein and down-regulation of Bax protein cause by CCL21. All the impact mentioned above was depend of the activation of CCR7.Conclusion CCL21/CCR7 axis enhances the migration and invasion of T24 cells, and promotes the ability of proliferation and anti-apoptosis of T24 cells.Part Three The relationship between CCR7 and VEGF-C expression in BUCAim Explore the relationship between CCR7 and VEGF-C expression in BUC and evaluate the impact of CCL21/CCR7 axis on VEGF-C expression of T24 cells.Method The relationship of CCR7 and VEGF-C expression level of BUC was estimated by Immunohistochemistry technique, RT-PCR was employed to measure the VEGF-C mRNA expression of T24 under the stimulation of CCL21. The VEGF-C protein expression was detected using western blot and the RNA bonding protein HuR was also detected under the stimulation of CCL21.Result There is an obvious relationship between expression of CCR7 and VEGF-C protein among 57 patients with BUC, the related coefficient was 0.309; VEGF-C mRNA expression appeared no change under the stimulation of CCL21, but the secretion of VEGF-C was increased under the stimulation of CCL21. The up-regulation of VEGF-C might be a result of post-transcript regulation of VEGF-C mRNA because HuR protein was also increased under the CCL21 treatment.Conclusion CCL21/CCR7 might increase the VEGF-C protein expression through a post-transcript regulation.