Effect Of MS-275 Combined With Cisplatin On ErbB3-overexpressing Human Bladder Transitional Cell Carcinoma,and Its Clinical Prospect

Part one:Effect of MS-275 combined with cisplatin on ErbB3-overexpressing human bladder transitional cell carcinoma,and its underlying mechanism.Objective:ErbB3 is a member of the epidermal growth factor receptor(ErbB/HER)family,which plays an important role in the development and progression of the tumor.Our previous study obtained the ErbB3-overexpressing human bladder urothelial carcinoma cell line by lentivirus transfection and found that the ErbB3-overexpressing human bladder urothelial carcinoma cell line was resistant to chemotherapeutic drugs such as cisplatin.Studies have shown that histone deacetylase inhibitor MS-275 can inhibit the expression of ErbB3 in ErbB2-overexpressing human breast cancer cells and promote apoptosis.In this study,we used MS-275 and cisplatin monotherapy and combination therapy to observe the effect of MS-275 and cisplatin on ErbB3-overexpressing human bladder urothelial carcinoma cell line.And to observe whether MS-275 could reverse ErbB3-overexpressing human bladder urothelial carcinoma cell line on cisplatin resistance and to study its underlying mechanism.Method:1?The survival rates of ErbB3-overexpressing group(T24-ErbB3,J82-ErbB3),lentivirus control group(T24-CV,J82-CV),control group(T24,J82)at different concentrations of MS-275,DDP monotherapy and combination therapy were analyzed by MTS assay.2?The effects of MS-275,DDP monotherapy and the combination therapy on the expression of PARP and Caspase8 in T24-ErbB3 cells at different time or different concentrations was detected by Western Blotting.3?Western Blotting was used to detect the effect of different concentrations of MS-275 on the expression of signal transduction-related proteins in T24-ErbB3 cells.Results:1?MTS results showed that the survival rates of ErbB3-overexpressing group,lentivirus control group,control group were gradually decreased upon treatment with increased concentration of MS-275,DDP.The effects of MS-275 and DDP combination therapy were better than monotherapy.There was statistically significant difference between ErbB3-overexpressing group and lentivirus control group(P<0.05).2?Western Blotting showed MS-275,DDP and MS-275 combined with DDP can increased apoptosis related proteins levels such as RARP and Caspase8.In combined group,the change of related proteins expression was more obvious.3?Western Blotting showed the expression of ErbB3?Akt and Survivin was down-regulated in combined group.Conclusion:MS-275 showed obviously antitumor effect on human ErbB3-overexpressing bladder transitional cell carcinoma T24/J82 cell line.MS-275 can enhance DDP on the antitumor effect of BTCC.The potential mechanism may be that MS-275 can down-regulate the expression of ErbB3,block ErbB3-PI3K/Akt-Survivin signaling pathway,and induce apoptosis.Part two:The expression of ErbB3,ErbB2 and Survivin in human bladder transitional cell carcinoma and its para-cancerous tissuesObjective:To discover the expression of ErbB2,ErbB3 and Survivin in human bladder urothelial carcinoma and their adjacent tissues,and to analyze the relationship between the expression of ErbB2,ErbB3 and Survivin in bladder urothelial carcinoma and the clinical or pathological stages.And to explore the correlation between ErbB2,ErbB3 and Survivin protein expression in bladder urothelial carcinoma.Method:We collected 20 cases of bladder transitional cell carcinoma tissues and their own para-cancerous tissues which were all confirmed by pathology specimens in March 20014-March 2016 in Dongfang hospital.The immunochemistry technologies were used for the detection.Positive tissue sections of the known breast cancer were selected as a positive control to ErbB2 and ErbB3,and colorectal tumor tissue to Survivin,PBS was slected instead of primary antibody as a negative control.The result was observed by two experienced pathologists in the unknown clinical and pathological data in accordance with the percentage of the number of positive cells in the slice of the results of grading.Results:In 20 cases of BTCC,12 patients showed positive expression of ErbB2,16 of ErbB3,14 of Survivin;In the normal bladder tissues,the ErbB2?ErbB3 and Survivin expression were3?6?2cases.Two goups were compared,the expression of ErbB2?ErbB3 and Survivin between BTCC group and para-cancerous group was statistically significant(P<0.05).The expression of ErbB2,ErbB3 and Survivin was not associated with clinical stage or pathological grade of bladder cancer.The expression of Survivin was correlated with that of ErbB3(r=0.771,P<0.01).Conclusion:The expression of ErbB3?ErbB2 and Survivin in BTCC tissues was higher than that in para-cancerous tissue significantly.In the bladder cancer,ErbB3 and Survivin expression has a strong correlation,the two may have synergies,common participation in the occurrence and development of bladder cancer and chemotherapy drug resistance process.ErbB3 is expected to be a new target for the treatment of bladder urothelial carcinoma.MS-275 has a bright future in the chemotherapy of ErbB3-overexpressing bladder cancer.