Mechanism Of Regulation Of LOXL2 To SNAIL In Cholangiocarcinoma Cells

Background: Cholangiocarcinoma(CCA)is one of the major diseases threatening human health.The incidence is increasing year by year and the early diagnosis is difficult.The operation rate of the first patient is less than 30% and the survival rate within 5 years after radical resection of cholangiocarcinoma is less than 10%.At present,the treatment method of cholangiocarcinoma is not too many,the overall prognosis of patients with cholangiocarcinoma is poor.The incidence of cholangiocarcinoma is higher in people over 40 years of age.The incidence of cholangiocarcinoma is slightly higher in males than in females.Cholangiocarcinoma threatens human life and causes a great economic and spiritual burden to families,society and countries.Therefore,it is urgent to find a new treatment method for cholangiocarcinoma.Invasion and metastasis are the main cause of death of cholangiocarcinoma.EMT is an important phenomenon in promoting invasion and metastasis of cholangiocarcinoma cells.SNAIL is an important factor that mediates the occurrence of EMT.SNAIL directly or indirectly promotes the occurrence of EMT by silencing E-Cadherin.LOXL2 is the key regulatory factor of EMT.It has been found that LOXL2 is involved in the post-translational modification of SNAIL protein,and this modification may depend on the K98 and K137 sites,while recently SUMO is discovered regulation of hundreds of nucleoproteins.The process and mechanism of LOXL2 regulating Snail is not clear.Based on the above studies and our previous experimental results,we propose the following scientific hypothesis: in cholangiocarcinoma cells,LOXL2 regulates the expression of SNAIL by mediating SUMOylation,and this regulation may be related to the K 98 and K 137 sites of SANIL.Objective: In cholangiocarcinoma cell lines RBE and QBC939,studying the process and mechanism of LOXL2 regulating SNAIL provides a new target for the prevention and treatment of cholangiocarcinoma,and provides new ideas for the treatment of other tumors.Method: In order to study the process and mechanism of LOXL2 regulating SNAIL,firstly we transfect EX-LOXL2?EX-LOXL2-EX-SENP?EX-LOXL2-Si-SUMO?EX-LOXL2-MU into RBE and QBC939 cell lines.Then we verify that LOXL2 regulates the expression of SNAIL by mediating SUMOylation,and this regulation is related to the K 98 and K 137 sites of SANIL by IF?CO-IP and Western blot.Result: ? In cholangiocarcinoma cell lines RBE and QBC939,when we transfected EX-LOXL2 plasmid,the expression level of SNAIL increased the and SNAIL Combine with SUMO.? In cholangiocarcinoma cell lines RBE and QBC939,when we transfected EX-LOXL2 plasmid and EX-SENP plasmids,the expression level of SNAIL decreased.when we transfected EX-LOXL2 and Si-SUMO,the expression level of SNAIL decreased at the same time.? In cholangiocarcinoma cell lines RBE,when we transfected EX-LOXL2 plasmid and MU-K98/K137 plasmid,the expression level of SNAIL decreased.Conclusion: in cholangiocarcinoma cell lines RBE and QBC939,LOXL2 positively regulates SNAIL by SUMOylation,and this process is related to the K98 and K137 sites of SNAIL.The results provide a new theoretical basis for the prevention and treatment of cholangiocarcinoma.