Study On The Effect And Mechanism Of XMU-MP-1 On Cholangiocarcinoma Cells Through Hippo Signaling Pathway

Objective Cholangiocarcinoma is one of the most common digestive tract cancers in the world,and the incidence rate is increasing year by year.Most patients with clinical cholangiocarcinoma have reached the middle and advanced stage,and the treatment effect is poor.Hippo signaling pathway plays a relatively stable role in mammalian cells,and its core components play an important role in regulating cell growth in various cells.This study explored the effect of XMU-MP-1 on the proliferation,invasion and migration of cholangiocarcinoma cells through Hippo signaling pathway,and explored its mechanism by detecting the expression level of downstream proteins.It is expected to provide more reference for the targeted treatment of cholangiocarcinoma in the future.Methods 1.Select CCK-8 experiment and adjust the concentration of XMU-MP-1 in complete medium(0,0.05,0.10,0.20,0.40,0.80,1.60,3.20μ M)according to relevant literature and reagent instructions,they were divided into 8 groups.Their effects on the proliferation of cholangiocarcinoma cells were studied at 24 h,48h and 72h;2.The appropriate concentration of XMU-MP-1 is selected according to the experimental results of CCK-8.According to different culture media,cholangiocarcinoma cells were divided into two groups: experimental group(containing XMU-MP-1 and fetal bovine serum)and control group(fetal bovine serum);3.Transwell invasion assay was used to detect the effect of XMU-MP-1 on the invasive ability of cholangiocarcinoma cells;4.The effect of XMU-MP-1 on the migration ability of cholangiocarcinoma cells was detected by cell scratch test;5.Western Blot was used to detect the expression of related factors LATS1,LATS2,YAP and TAZ in Hippo signal pathway downstream of XMU-MP-1 action site in the two groups.Results 1.CCK-8 experiment found that within the concentration range of this experiment,the proliferation activity of cholangiocarcinoma cells increased with the increase of XMU-MP-1culture concentration and culture time;2.Transwell invasion experiment found that XMU-MP-1 can effectively improve the invasion ability of cholangiocarcinoma cells;3.Cell scratch test showed that XMU-MP-1 could effectively enhance the migration ability of cholangiocarcinoma cells;4.The results of Western blot showed that after XMU-MP-1 treatment,the expression of LATS1 and LATS2 decreased,and the expression of YAP and TAZ increased.Conclusion 1.XMU-MP-1 can effectively enhance the proliferation,invasion and migration of cholangiocarcinoma cells;2.XMU-MP-1 can promotes the proliferation,invasion and migration of cholangiocarcinoma cells by inhibiting Hippo pathway,down regulating the expression of LATS1 and LATS2 proteins and up regulating the expression of YAP and TAZ proteins.