Combination Chemotherapy With Zyflamend Reduced The Acquired Resistance Of Bladder Cancer Cells To Cisplatin Through Inhibiting NFκB Signaling Pathway

Background:Urinary bladder carcinoma is one of the most common malignant tumors in urinary system,which seriously affects human life and health.Bladder cancer is treated mainly by surgery combined with chemotherapy,including intravesical instillation chemotherapy or adjuvant systemic chemotherapy before or after radical surgery.cisplatin based chemotherapy is the main therapy for bladder cancer.Chemotherapy tolerance is one of the most important factors affecting the poor prognosis of tumor patients.However,the mechanism of acquired chemotherapy resistance remains unclear.Inflammatory factor activation may be one of the important causes of drug resistance and recurrence of tumor.We intend to detect the expression of inflammatory-related signaling pathway by constructing a cisplatin-tolerant bladder cancer cell line.At the same time,the potential effects of a polyherbal preparation,Zyflamend,on cisplatin-resistant bladder cancer cells were also investigated.Methods:Firstly,we construct the cisplatin tolerant bladder cancer cell line(T24R).T24 cells were cultured in cisplatin for 10 months.The drug resistance of T24R cell line was confirmed by MTT assay.Cell proliferation and NF κ B signaling pathway related gene expression were detected by Western Blot.We treated these cells with different concentrations of Zyflamend to detect proliferation and the activity of NF κ B signaling pathway.In order to detect the synergistic effect between Zyflamend and cisplatin,T24R cells were treated withZyflamend or cisplatin alone or in combination.The apoptotic effect was evaluated by Annexin V/PI double staining,and the levels of cell cycle and anti-apoptosis-related proteins were detected by Western Blot.Finally,the nude mice were heterograft with T24R cells.We treated with cisplatin and Zyflamend alone or in combination to measure the size and weight of tumor tissue,and to collect tumor tissue samples.In addition,the indexes of proliferation and apoptosis were detected by immunohistochemistry.Results:Compared with parent T24 cells,NF κ B signaling pathway was activated in cisplatin-resistant T24R cells.Zyflamend inhibited the growth of T24,T24R and another bladder cancer cell line J82 in a concentration-dependent manner.In the mechanism,Zyflamend inhibits NF κ B-mediated cell proliferation,survival and invasion,and induces cell apoptosis.In addition,Zyflamend signiflicantly increased the sensitivity of T24R and J82 cells to cisplatin therapy.These results were also confirmed in the T24R xenograft tumor model.When treated with Zyflamend and cisplatin,the tumor volume decreased,the proliferation of cells decreased,and the expression of ReIA and its downstream target protein MMP9 decreased.Conclusion:Our results suggest that cisplatin-resistant bladder cancer cells activate the NF κB signaling pathway.Zyflamend can inhibit cell proliferation and induce apoptosis by targeting NFκB signaling pathway,and treat cisplatin resistance of bladder cancer.