Study On The Anti Esophageal Squamous Cell Carcinomaactivity And Mechanism Of Dandelion Root Extract

OBJECTIVE: To study the effect of water dandelion root extract?DRE?on the growth of esophageal squamous cell carcinoma?ESCC?and the role of CBS / H₂S system in ESCC.And objective to investigate the inhibitory effect of DRE on esophageal squamous cell carcinoma and its mechanism.METHODS: The effects of DRE on the growth,proliferation,migration,invasion and apoptosis of ESCC were detected by MTT,EdU,cell scratch,Transwell and apoptosis experiments,Esophageal pingsan was the positive control;The expression of cystathionine-?-lyase?CSE?and cystathionine-?-synthase?CBS?in esophageal squamous cell carcinoma cells and esophageal normal epithelial cells was detected by Western Blot,and their H₂S levels were measured by H₂S fluorescence probe method;In order to evaluate the role of CBS / H₂S system in esophageal squamous cell carcinoma,KYSE450 and NEC cells were transfected with CBSsiRNA to detect the effect of CBS down-regulation on the growth,proliferation,migration and invasion of esophageal squamous cell carcinoma cells;H₂S fluorescence probe and Western Blot were used to detect the effect of DRE on the level of H₂S and the expression level of CBS and CSE protein in ESCC cells.In order to evaluate the anti esophageal squamous cell carcinoma effect of DRE,a model of esophageal squamous cell carcinoma transplantation was constructed to detect the inhibition of DRE on tumor growth in vivo.RESULTS:1.MTT,EdU,cell scratch,Transwell and apoptosis experiments showed that DRE can effectively inhibit the growth,proliferation,migration and invasion of esophageal squamous cell carcinoma KYSE450 and NEC cells,and induce apoptosis.Moreover,the toxicity of DRE to esophageal normal epithelial cells was significantly lower than that of positive control esophageal pingsan.2.Western Blot showed that there was no significant difference in the expression of CSE between esophageal squamous cell carcinoma and esophageal normal epithelial cells,while the expression of CBS in KYSE450,NEC,EC9706 cells was higher than that in NE-3 cells.3.The results of H₂S fluorescence probe showed that the endogenous H₂S released of KYSE450 and NEC cells was significantly higher than that of NE-3 cells.4.After CBSsiRNA was used to down regulate the expression of CBS in KYSE450 and NEC cells,the release of H₂S in KYSE450 and NEC cells was significantly reduced,and the growth,proliferation,migration and invasion of cells were inhibited.5.DRE can inhibit the expression of CBS and the release of H₂S in KYSE450 and NEC cells in a dose-dependent manner,but it has no significant effect on the expression of CSE.6.DRE can significantly inhibit the growth of tumor in vivo.Compared with the positive control group,the effect of DRE is slightly better than that of the positive control group,and it has no toxic and side effects on the body.CONCLUSIONS:1.DRE can effectively inhibit the growth,proliferation,migration and invasion of KYSE450 and NEC cells,and induce apoptosis.The toxicity of WEDR to esophageal normal epithelium cells is significantly lower than that of positive control esophageal pingsan.2.The level of H₂S in ESCC cells depends on the level of CBS.The high expression of CBS / H₂S system may promotes the development of ESCC.3.DRE may exert its anti esophageal cancer activity by down regulating CBS / H₂S system.4.DRE can inhibit the growth of tumor in vivo and may has no toxic and side effects on the body.