Disulfiram Inhibits The Expression Of ALDH1A1 In Bladder Cancer Cells And Enhances Its Cisplatin Sensitivity

Objective: Worldwide,nearly 150,000 people die of bladder cancer each year.At present,for advanced bladder,there are mainly endoscopic and open surgery,local or systemic immunotherapy,radiotherapy and chemotherapy,etc.,although these treatment strategies have improved the treatment effect and prolonged the survival and life treatment of bladder cancer patients.However,the disease is still not completely cured and has a high risk of recurrence,and the prognosis is not ideal.In recent years,scholars have found that there are a group of undifferentiated bladder tumor cells,which are called bladder cancer stem cells.They believe that tumor recurrence and poor prognosis are related to bladder cancer stem cells.Many studies on bladder cancer stem cells have been published,and CD44,Ck5,CD47,ALDH and other molecules are considered to be molecular markers of cancer stem cells.Based on these molecular markers of bladder cancer stem cells,new treatments for bladder cancer are provided.Ideas and methods.Reusing old medicines for new purposes,such as using old medicines to achieve cancer suppression in clinical treatment of tumors,is a very attractive and exciting area.Disulfiram is a good example of this.Disulfiram is re-used as a potential anticancer drug due to its unique known properties,low cost,low side effects and high selectivity for different cancers and synergistic activity with other drugs.Moreover,one of the mechanisms by which disulfiram inhibits tumor cells is as an inhibitor of ALDH.This experiment explored the effect of disulfiram on the expression of ALDH1A1 in bladder tumors and its potential mechanism of enhancing the sensitivity of tumor cells to cisplatin.Methods: The experiment was divided into a control group and a disulfiram(DSF)group.Search the relevant database to query the expression of ALDH1A1 bladder cancer specimens.The mRNA of EJ cells and SV-HUC-1 cells were extracted,and the expression of ALDH1A1 was detected.The effect of DSF on the colonization ability of bladder tumor cells was examined by colony formation assay.The RNA of bladder tumor cells in DSF intervention group and control group were extracted,and the expression of ALDH1A1 and other dry related genes in tumor cells were detected by real-time fluorescent quantitative PCR after real-time quantitative PCR.The effect of DSF on the migration ability of bladder tumor cells was examined by scratch test at 0H and 24 H after DSF intervention.The effect of flow cycle on the cell cycle of bladder tumors after DSF intervention.The effect of DSF on the sensitivity of tumor cells to cisplatin was detected by CCK8 method and flow cytometry.Results: The number of cloned cells in the bladder cancer cells of the DSF group was significantly lower than that of the control group.After the clones of the control group and the DSF group were formed,the pellets were stained with crystal violet.After dissolving the crystal violet,the absorbance at 570 nm of the two groups was 0.884�0.022 and 0.490�0.027,respectively,and the difference was statistically significant(P<0.001).Compared with normal urothelial cells SV-HUC-1,the expression of ALDH1A1 in bladder cancer EJ cells was significantly increased,and the difference was statistically significant(P<0.001).Compared with the control group,the expression level of ALDH1A1 in DSF group was significantly down-regulated.The expression of stem cell genes such as NOTCH2 and SHH was also down-regulated,and the difference was statistically significant(P<0.05).Compared with the control group,the migration ability of bladder tumor cells after DSF intervention was significantly decreased.Compared with the control group,the number of bladder tumor cells in the G1 phase increased significantly after DSF intervention,while the number of cells in the S phase decreased.The cisplatin combined with DSF and cisplatin alone were used to treat bladder tumor cells for 24 h.The median cell killing concentrations of cisplatin were 8umol and 32 umol,respectively,and the difference was statistically significant(P<0.05).Bladder tumor cells after DSF intervention become sensitive to cisplatin.Conclusion: Bladder cancer tissue has higher ALDH1A1 expression than adjacent tissues.The expression of ALDH1A1 in bladder cancer cells is higher than that in normal urothelial cells.As an inhibitor of ALDH,disulfiram can affect the expression of ALDH1A1 in bladder tumor cells,affecting the migration,clone formation ability and cell cycle phenotype of bladder tumor cells,it also enhances the sensitivity of bladder tumor cells to cisplatin.