The Study Of CYLD And Livin In Chemosensitivity Of Bladder Cancer

BackgroundBladder cancer is the most common type of cancer in western countries and Asian countries.The prevalence and mortality of bladder cancer are increasing year by year.It is the fourth most common cancer in the United States and the second most common malignancy in males.It ranks the first in Chinese uriary system tumors and its incidence has been increasing in recent years.Although the early detection of bladder cancer means has become universal and patients 5-year survival rate has already get to 80%,but its 5-year recurrence rate of more than 50%,and there will be 10-20%progress into invasive bladder cancer.Gemcitabine is a first-line chemotherapeutic agent for bladder cancer and has been widely used in clinic.However,chemotherapy resistance is still an unavoidable problem..So,to overcome chemoresistance and improve Gemcitabine chemotherapeutic efficacy,novel molecular marker for chemosensitivity has been highly desirable.ObjectivesIn this study,we first invest the relationship of CYLD combined with Livin in bladder cancer,and then by stable transfection CYLD and Livin to discover CYLD combined with Livin in improving gemcitabine-induced cytotoxicity,autophagy and apoptosis.In order to provide a new experimental basis for exploring new molecular markers to enhance chemosensitivity of tumor cells and possible mechanisms of action to accelerate tumor cell killing and to improve the traditional chemotherapy regimen.Materials and methods(1)Cultured bladder cancer cells(253J,T24),transfected with CYLD and Livin,and the expression levels of CYLD and Livin in each cell line were detected by Western blot and reverse transcription-PCR.(2)The inhibitory effects of CYLD and Livin on bladder cancer cells were detected by cell viability assay(CCK-8),and then detect the expression of CYLD and Livin by Western blot.The expression of CYLD and Livin in each bladder cancer cell line,the expression of NF-kappaB signal pathway and the expression of autophagy and apoptosis related proteins in T24 and 253J cells were detected by Western blot.(3)Transwell invasion assay was used to detect the invasion and migration ability of CYLD and Livin in invasive bladder cancer cell lines(253J,T24).The expression of autophagy and apoptosis was detected by Western blot,and the mechanism of CYLD and Livin on the chemosensitivity of bladder cancer cells was investigated.(4)Using bladder cancer cell lines 253J and T24 to establish animal model of bladder cancer(mouse subcutaneous tumor model)to study the effect of various drugs in vivo treatment of tumors;intraperitoneal injection of drugs injected into mice and grouped,(DMSO),CYLD group,Livin group and CYLD + Livin group.The effect of each group on the growth of subcutaneous tumor of bladder cancer was observed,and the inhibition of tumor cell proliferation was analyzed.Results(1)CYLD and Livin could increase the chemotherapy and sensitivity of bladder cancer cells,while CYLD combined with Livin could significantly increase the cytotoxicity induced by gemcitabine,and increase the expression of CYCL and Livin in bladder cancer cells Sensitivity.(2)CYLD and Livin could inhibit invasion and migration of invasive bladder cancer cell lines 253J and T24,and CYLD combined with Livin could inhibit the invasion and migration of bladder cancer cells.(3)Western blotting detection of muscle invasive bladder cancer cell lines 253J,T24,found that bladder cancer cell line gemcitabine chemotherapy sensitivity may be related to inhibition of cell autophagy protein LC3,P62,promote apoptosis protein Caspase3,Smac,And found that CYLD can regulate Livin through the NF-kB signal pathway,thereby enhancing chemotherapy sensitivity of cancer cells gemcitabine.(4)The results of in vivo experiments showed that CYLD + Livin could inhibit the growth of subcutaneous tumors and inhibit the proliferation of tumor cells.Conclusions(1)Increased expression of CYLD and downstream NF-kB signaling pathway is closely related to the progression of bladder cancer and suppresses autophagy of bladder cancer cells and promotes cancer cell apoptosis.(2)Livin can inhibit the invasion and migration of bladder cancer cell lines,and by CYLD regulation and control.(3)CYLD combined with Livin can significantly increase the cytotoxicity induced by gemcitabine and regulate autophagy and apoptosis in bladder cancer cells.This combination therapy provides an effective new treatment for overcoming the chemotherapy resistance of bladder cancer.