The Her-1/Her-2Expression In Bladder Cancer And Research Of The Effecting Of Afatinib On Bladder Cancer T24Cells

Part one The correlation between expression of HER-1/HER-2in bladder transitional cell carcinoma(BTCC) and clinical significanceAim Explore the situation of expression of HER-1/HER-2in BTCC and evaluate the correlation between HER-1/HER-2with the parameters of BTCC such as tumor stage and grade.Method The expression level of HER-1/HER-2was estimated by Immunohistochemistry technique. And the influence factors were also tested.Result The positive rate of HER-1expression among56patients was55.4%, which was distinct higher than that in normal bladder tissue (10%); the positive rate of HER-2expression among56patients was37.5%, which was distinct higher than that in normal bladder tissue (0%); The expression level of HER-1was obvious greater in high stage group, high grade and relapse group than that in low stage group, low grade group and primary group. The expression level of HER-2was obvious greater in high stage group and relapse group than that in low stage group and primary group.Conclusion The expression of HER-1/HER-2was up-regulated in BTCC, and their expression has the positive correlation. Part two The influence of Afatinib on T24cells'biologic behaviorAim Estimate the influence of Afatinib on T24cells'biologic behavior such as invasion, proliferation and apoptosis.Method The T24cell was grouped depend on the concentration of Afatinib treatment (0,1,5,10,20umol/L). MTT was employed to evaluate the impact of Afatinib to T24proliferation; FCM was employed to evaluate the impact of Afatinib to T24with apoptosis ability; transwell analysis was used to detect the effect of Afatinib to T24invasion. The invasion associated protein MMP-2and MMP-9, the apoptosis associated protein Bcl-2and Bax was detected by western blot. Check the ERK1/2and Akt to explore the mechanism of the Afatinib in bladder cancer.Result The T24cells showed more powerful repression of proliferation and invasion under the treatment of Afatinib. Afatinib could regulate the expression of MMP-2/MMP-9and BAX/Bcl-2proteins. Afatinib could promote the apoptosis ability of T24.Conclusion Afatinib inhibition of T24cell proliferation and invasion and promotes apoptosis in T24cells. Part three Afatinib influencing chemotherapy sensibility that cisplatin treat T24cellAim To research Afatinib influencing chemotherapy sensibility that cisplatin treat T24cellMethod First we prepared the T24-ADM cell, then observation of Afatinib combined with cisplatin on the inhibition of T24by MTT. Using the MTT to test sensibility of cisplatin before and after Afatinib used in T24-ADM cell. Using the PCR to test MDR-1before and after Afatinib used in T24-ADM cell and using the western-blot to test p-gp before and after Afatinib used in T24-ADM cell.Result Combining Afatinib and cisplatin could of inhibit T24cell proliferation better than with cisplatin alone; cisplatin could of inhibit T24cell proliferation better if the Afatinib was used. Afatinib can inhibit the mdr-1gene and P-gp protein.Conclusion Afatinib and cisplatin showed a synergistic effect in the inhibition of T24cell proliferation. Cisplatin could of inhibit T24cell proliferation better if the Afatinib was used. Afatinib can inhibit the mdr-1gene and P-gp protein.