The Inhibit Effect Of Diosgenin To The Bladder Cancer Cells(T24 And 5637)

Background:Bladder cancer(BC)is one of the most common tumors over the world,and its incidence and mortality have occupied great attention in the past decades.Approximately 357,000 new individuals are occurring around the world and about 145,000 people dying from this disease each year.Among them,approximately 70%BCa patients are non-muscle-invasive bladder cancer(NMIBC).According to the SEER-Medicare-linked database of the National Cancer Institute(USA),almost 40%of patients with high-risk NMIBC experienced recurrence without progression.Another 33%experienced disease progression,40%of whom died as a result of the progression.To reduce the high rate of recurrence,intravesical therapy has been demonstrated to be the most effective therapy for intermediate-high-risk NMIBC.For muscle-invasive BCa,now the gold standard treatment is radical cystoprostatectomy[5],but this therapeutic approach still has many unfavorable outcomes.Therefore,an effective strategy for preventing the progression of BCa is urgently needed.Diosgenin,a naturally occurring steroidal saponin.It was found in variety of plants including Trigonella foenum graecum and roots of wild yam,is a well-known precursor of various synthetic steroidal drugs that are extensively used in the pharmaceutical industry.Diosgenin has been used in traditional medicine for old patients with hyperlipidemia,hypercholesterolemia and atherosclerotic disease,It has been demonstrated that diosgenin inhibits proliferation and promotes apoptosis in a wide variety of tumor cells of human colon,osteosarcoma,leukemia,erythroleukemia,breast and liver.Therefore,diosgenin may have a new therapeutic potential in anti-metastatic therapy for bladder cancer.Apoptosis,or programmed cell death,is critical for the normal development,function and homeostasis of multi-cellular organisms.Biochemical events lead to characteristic cell changes(morphology)and death.Cell apoptosis was also considered to be an efficient way which may be wipe out the precancerous and cancerous cells,and the reduction of apoptosis is one of the basic mechanisms for tumorigenesis.Therefore,it is a promising strategy for enhancing the anticancer potential of several anticancer drugs which are able to induce,or enhance,the extent of apoptosis.The expression of some proteins(Bcl-2 and Bax)can obviously affect the apoptotic process,which has been clearly demonstrated.A lot of evidence supports that cell cycle arrest and apoptosis play an improtant role in cell proliferation.The transition from one cell phase to another is an well-ordered process,and cell cycle control is the major regulatory mechanism underlying cell growth,which is regulated by cyclin-dependent kinases(CDKs)and their cyclin partners.Epithelial-to-mesenchymal transition(EMT)is an important role in the metastasis process that can promote migration.Organ development,tumor growth and cancer progression.Meanwhile,an accumulating body of evidence revealed that EMT acts as a major driver of tumor invasion,the expression of protein markers,such as N-cadherin,vimentin and ?-catenin,are positive for majority of cases of cancer.Although the effects of Diosgenin on antiproliferative and apoptotic activity have been documented in various human cancer cells.But its therapeutic efficacy and the underlying mechanisms in bladder cancer cell is still unknown.The aim of this study is to investigate the mechanism of Diosgenin on proliferation,apoptosis and metastasis in bladder cancer cells,by using human bladder cancer cell lines(5637 and T24).Objective Diosgenin,a steroidal saponin extracted from Trigonella foenum graecum,was a drug for old patients with hyperlipidemia,hypercholesterolemia and atherosclerotic disease in China,meanwhile,it also been found an anticarcinogenic properties in several tumor cell lines.However,the effect of Diosgenin on bladder cancer remains unclear.The aim of the study was to examine the effect of Diosgenin in human bladder cancer T24 and 5637 cells.MethodsMTT assay was used to determine the effects of Diosgenin on the viability of these cells.The cell cycle arrest was detected through propidium iodide(PI)staining.The induction of apoptosis was determined through Annexin V staining.We used the methords of PCR and Western blot to detect the expression of the mRNA and protein of Bcl-2,Bax,Vimentin,etc.in bladder cancer T24 and 5637 cells,and the transwell assay and scratch test was used to testify the cell migration efficiency.Results and ConclusionIn present study,it was found that Diosgenin could obviously inhibit proliferation of the bladder cancer cell lines demonstrated by MTT assay and Clonogenic survival assay.In a concentration range of 0 to 30 ?g/mL,the inhibitory effects of Diosgenin on proliferation of the bladder cancer cell were increased in a dose and time-dependent manner(P<0.01).Flow cytometry indicated that Diosgenin could inhibit the DNA synthesis at G0/G1 phase(P<0.01).The apoptosis rates of the the bladder cancer cells treated with Diosgenin were increased significantly compared with the control group(P<0.01).All the DNA frequency histogram in Diosgenin-treated groups showed obvious hypodiploid peak and with the increase of Diosgenin concentrations.Wound healing assay and transwell assay shown that the cell migration capacity were markedly inhibited.It was also found that Diosgenin could up-regulate the expression of Bax and down-regulate the expression of Bcl-2,CDK2/4/6,N-cadherin,Vimentin,etc.It was concluded that Diosgenin could inhibit proliferation of bladder by interrupting the cell cycle at G0/G1 restriction point via decrease the expression of CDK2/4/6,Cyclin D1 and induce apoptosis of T24 and 5637 cells via reducing the ratio of Bcl-2/Bax.It also could effect bladder cancer cell migration by downregulation the quantity of N-Ca/Vimentin,etc.These findings reveal that diosgenin may have the therapeutic potential on bladder cancer.