The Search Of The New Pattern Pathway In Bladder Cancer Cell And The Influence Of Formononetin On The Apoptosis Signaling Pathways Of Bladder Caner Cell

Background:Bladder cancer(BC)is one of the most common human malignancies affecting the urinary system,which is caused by multi-factors,and short of effective diagnostic methods and drugs.SUMOylation is an essential posttranslational modification characterized by covalent conjugation of SUMO proteins to many different proteins,which emerged as a major regulatory mechanism for a variety of cellular processes including gene expression,DNA repair etc.Dysregulation of sumoylation in various types of cancer is closely associated with tumorigenesis and drug sensitivity.But the role and molecular mechanism of sumoylation in BC remains unclear.At present,operation is the mainly treatment.As the indispensable treatment for end-staged BC,chemoradiotherapy led to a heavy economic burden and pain for the patients.So,discovering a novel antineoplastic drug with more effects and less toxicity is very important.Recent studies identified that Formononetin could inhibit proliferation and induce apoptosis of several tumor cells such as prostate cancer and breast cancer cells.Objective:In order to elucidate the role of sumoylation on bladder cancer with database analysis(GEO and TCGA),immunohistochemistry,shRNA,RNA-seq etc;and identify the mechanism of anti-BC with formononetin through up-regulating apoptosis related factor(Bax)in vitro.Methods:On the basis of GEO and TCGA databases,immunohistochemistry,shRNA,RNA-seq and so on,the role of sumoylation in BC was presented;on the basis of MTT and FCM assay,the role of formononetin in the effect of anti-cancer was presented;and the expression of Bax changed with the different concentration of formononetin according to Western blot and qPCR.Result:The results of GEO and TCGA databases showed that SUMO pathway is overactive in BC.Moreover,immunohistochemistry also show that SUMO subtypes and SUMO ligase UBC9 were overexpression in different malignancy of bladder cancer tissue.Additionally,knockdown of UBC9 with shRNA in bladder cancer cells could inhibit cell proliferation,with many pathways associated with growth and with proliferation subsequently changed,the result of MTT assay showed that formononetin could effectively inhibit proliferation of BC cell,dependent on the concentration;FCM assay showed that formononetin could inhibit proliferation of BC cell mainly through inducing cellular apoptosis.Moreover,the expression of Bax was rising along with the concentration of formononetin according to Western blot and qPCR.Conclusion:Sumoylation is crucial for the initiation and development of BC.And it may be a valuable target for diagnosis and treatment of BC in the future;Formononetin could inhibit proliferation of BC cell and induce cell apoptosis through up-regulated expression of Bax possibly.